Search

Objective: To investigate the role of RNA m6A methylation in mediating cerebellar dysplasia through analyzing the phenotypes of the mouse cerebella and the expression of several key m6A regulators upon hypobaric hypoxia treatment. Methods: Five-day old C57/BL6 mice were exposed to hypobaric hypoxia for 9 days. The status of mouse cerebellar development was analyzed by comparing the body weights, brain weights and histological features. Immunostaining of cell-type-specific markers was performed to analyze the cerebellar morphology. Real-time PCR, Western blot and immunohistochemical staining were performed to detect the expression of key m6A regulators in the mouse cerebella. Results: Compared with the control, the body weights, brain weights and cerebellar volumes of hypobaric hypoxic mice were significantly reduced ( P< 0.01). The expression of specific markers in different cells, including NeuN (mature neuron), Calbindin-D28K (Purkinje cell) and GFAP (astrocyte), was decreased in hypobaric hypoxic mouse cerebella ( P< 0.01), accompanied with disorganized cellular structure. The expression of methyltransferase METTL3 was significantly down-regulated in the cerebella of hypobaric hypoxic mice ( P< 0.05). Conclusions: Hypobaric hypoxia stimulation causes mouse cerebellar dysplasia, with structural abnormalities in mature granular neurons, Purkinje cells and astrocytes. Expression of METTL3 is decreased in hypobaric hypoxic mice cerebellum compared with that of normobaric normoxic mice, suggesting that its mediated RNA m6A methylation may play an important role in hypobaric hypoxia-induced mouse cerebellar dysplasia.

Objectives: To compare the repair effects of different doses of human umbilical cord mesenchymal stem cells (hUC-MSCs) on white matter injury (WMI) in neonatal rats., Methods: Two-day-old Sprague-Dawley neonatal rats were randomly divided into five groups: sham operation group, WMI group, and hUC-MSCs groups (low dose, medium dose, and high dose), with 24 rats in each group. Twenty-four hours after successful establishment of the neonatal rat white matter injury model, the WMI group was injected with sterile PBS via the lateral ventricle, while the hUC-MSCs groups received injections of hUC-MSCs at different doses. At 14 and 21 days post-modeling, hematoxylin and eosin staining was used to observe pathological changes in the tissues around the lateral ventricles. Real-time quantitative polymerase chain reaction was used to detect the quantitative expression of myelin basic protein (MBP) and glial fibrillary acidic protein (GFAP) mRNA in the brain tissue. Immunohistochemistry was employed to observe the expression levels of GFAP and neuron-specific nuclear protein (NeuN) in the tissues around the lateral ventricles. TUNEL staining was used to observe cell apoptosis in the tissues around the lateral ventricles. At 21 days post-modeling, the Morris water maze test was used to observe the spatial learning and memory capabilities of the neonatal rats., Results: At 14 and 21 days post-modeling, numerous cells with nuclear shrinkage and rupture, as well as disordered arrangement of nerve fibers, were observed in the tissues around the lateral ventricles of the WMI group and the low dose group. Compared with the WMI group, the medium and high dose groups showed alleviated pathological changes; the arrangement of nerve fibers in the medium dose group was relatively more orderly compared with the high dose group. Compared with the WMI group, there was no significant difference in the expression levels of MBP and GFAP mRNA in the low dose group ( P >0.05), while the expression levels of MBP mRNA increased and GFAP mRNA decreased in the medium and high dose groups. The expression level of MBP mRNA in the medium dose group was higher than that in the high dose group, and the expression level of GFAP mRNA in the medium dose group was lower than that in the high dose group ( P <0.05). Compared with the WMI group, there was no significant difference in the protein expression of GFAP and NeuN in the low dose group ( P >0.05), while the expression of NeuN protein increased and GFAP protein decreased in the medium and high dose groups. The expression of NeuN protein in the medium dose group was higher than that in the high dose group, and the expression of GFAP protein in the medium dose group was lower than that in the high dose group ( P <0.05). Compared with the WMI group, there was no significant difference in the number of apoptotic cells in the low dose group ( P >0.05), while the number of apoptotic cells in the medium and high dose groups was less than that in the WMI group, and the number of apoptotic cells in the medium dose group was less than that in the high dose group ( P <0.05). Compared with the WMI group, there was no significant difference in the escape latency time in the low dose group ( P >0.05); starting from the third day of the latency period, the escape latency time in the medium dose group was less than that in the WMI group ( P <0.05). The medium and high dose groups crossed the platform more times than the WMI group ( P <0.05)., Conclusions: Low dose hUC-MSCs may yield unsatisfactory repair effects on WMI in neonatal rats, while medium and high doses of hUC-MSCs have significant repair effects, with the medium dose demonstrating superior efficacy.

Objective: To observe the effects of transcutaneous auricular vagus nerve stimulation (taVNS) on the motor function and the expression of glial fibrillary acidic protein (GFAP) and microtubule associated protein 2 (MAP2) in cerebral ischemic penumbra of rats with middle cerebral artery occlusion (MCAO) and explore the mechanism of taVNS in the improvement of motor function in MCAO rats., Methods: A total of 48 male SD rats were randomized into a sham-operation group, a model group, a transcutaneous auricular non-vagus nerve stimulation (tnVNS) group and a taVNS group, with 12 rats in each group. The suture-occluded method was adopted to prepare MCAO rat model. The auricular rim was stimulated in the tnVNS group and the concha stimulated in the taVNS group, 2 mA in intensity, 10 Hz in frequency, 30 min each time, once a day, for 14 days consecutively. The nerve functional assessment was recorded in each group. The expressions of nicotinic acetylcholine receptor (α7nAchR) in the cerebral ischemic penumbra and the spleen were detected by using Western blot. With the immunofluorescence, the expressions of GFAP and MAP2 were detected., Results: After modeling, compared with the sham-operation group, the nerve functional score was increased in the model group, the tnVNS group and the taVNS group ( P <0.01), suggesting the success of modeling. After treatment, the score was increased in the model group ( P <0.01) as compared with the sham-operation group. Compared with the model group, the neurological deficit score was reduced in the taVNS group ( P <0.01). Compared with the sham-operation group, GFAP expression was increased and MAP2 expression was reduced remarkably in the cerebral ischemic penumbra in the model group ( P <0.05). In comparison with the model group, GFAP expression was reduced, while MAP2 expression was increased remarkably in the cerebral ischemic penumbra in the taVNS group ( P <0.05). There were no significant differences in the abovementioned indexes between the model group and tnVNS group ( P >0.05). The differences in the expression of α7nAchR in the cerebral ischemic penumbra and the spleen had no statistical significance among groups ( P >0.05)., Conclusion: TaVNS is effective on neuroprotection in MCAO rats, which may be related to its function of inhibition of GFAP expression and promotion of MAP2 expression in the ischemic penumbra.

Objective: To explore the effects of intrauterine infection on early growth and neurobehavioral development in neonatal rats. Methods: Escherichia coli (E. coli) was inoculated into uterine cervix of pregnant rats with gestation of 15 d to establish the intrauterine infection model, and the effect on the delivery of pregnant rats was observed. The neonatal rat brain tissue was stained with Hematoxylin-Eosin and the cerebral white matter damage was assessed. Immunohistochemical staining and Western blot analysis were performed to evaluate the expression of glial fibrillary acidic protein (GFAP), 2', 3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) and neurofilament (NF) in pup brains. Birth weight and early growth development indices were monitored,and neurobehavioral tests were performed to access the change of neurobehavioral development in neonatal rats. Results: The white blood cell count increased significantly in the uterus and placenta of the pregnant rats after intrauterine E. coli infection and no significant impact was observed on the delivery of pregnant rats. Weak staining and focal rarefaction of cerebral white matter from rats at P7 in intrauterine infection group were observed. The expression of GFAP markedly increased ( P <0.05) in infection group, while the level of CNPase and NF in pup brains at P7 significantly decreased ( P <0.05 or P <0.01). Compared with control group, the neonatal rats in infection group had lower birth weight and slower weight gain during the suckling period ( P <0.05 or P <0.01), and the completion times of ear opening, eye opening, surface righting, negative geotaxis, acoustic startle and swimming test in infection group were significantly delayed ( P <0.05 or P <0.01). Conclussion: Intrauterine infection in pregnant rats can induce cerebral white matter damage and retardation of early growth and neurobehavioral development in neonatal rats.

Objective To observe the effects of microRNA-7 (miR-7) on intracerebral hemorrhage-induced brain injury in rats and explore the underlying mechanism. Methods Seventy-five SD rats were used to establish intracerebral hemorrhage model through injection of VII collagenase into the pallidum. These rats were then divided into control, miR-7 agomir, agomir control, miR-7 antagomir and antagomir control groups, containing 15 animals in each group. On day 2 after modeling, rats were injected with 10 μL of physiological saline, miR-7 agomir, agomir control, miR-7 antagomir and antagomir control via the lateral ventricle, respectively. On day 7 after modeling, the neurological function score was evaluated, and then all rats were killed to obtain brain tissue. HE staining was conducted to observe the pathological changes of cerebral tissue. Brain water content was examined by the dried and wet mass. The expressions of miR-7, glial fibrillary acidic protein (GFAP) and epidermal growth factor receptor (EGFR) in the cerebral tissue around hemotomas were detected using real-time quantitative PCR. Western blotting was used to analyze the levels of GFAP, EGFR, signal transducers and activators of transcription 3 (STAT3) and phosphorylated STAT3 (p-STAT3) in the cerebral tissue around hemotomas. In addition, bioinformatics was used to predict the binding of miR-7 to EGFR. Both wild type luciferase reporter gene vector and corresponding mutant vector were constructed and then transfected into HEK293T cells along with miR-7 mimic for detecting the luciferase activity. Results In comparison with control group, miR-7 agomir markedly increased miR-7 level, alleviated the pathological impairment of cerebral tissues, reduced neurological function score and brain water content, and attenuated the expression levels of GFAP and EGFR and p-STAT3/STAT3 ratio in the cerebral tissue around hemotomas. An opposite effect was observed in response to miR-7 antagomir. However, their negative controls had no impact on the above indicators. There was a binding site between 3'-untranslated region within the EGFR and miR-7. Moreover, miR-7 mimic markedly decreased the luciferase activity of the reporter gene of wild type, but not its mutant. Conclusion The miR-7 can inhibit the EGFR/STAT3 signaling pathway to antagonize astrocyte activation, leading to the protection against brain injury after intracerebral hemorrhage in rats.

Objective: To identify the clinical and genetic characteristics in 43 Chinese children diagnosed with type Ⅰ Alexander disease (AxD). Method: Forty-three type Ⅰ AxD cases identified by glial fibrillary acidic protein (GFAP) gene mutations in Peking University First Hospital from 2005 to 2016 were followed up. The data of medical history, physical examination and magnetic resonance imaging (MRI) were collected. All these patients were followed up in December 2010, Febury 2012, June 2014 and January 2016, respectively. Result: Forty-three patients were genetically confirmed as type I AxD and the median age at the last visit was 11.71 years (10.27, 13.15). The characteristic clinical manifestations of these type Ⅰ AxD patients were developmental delay (79%, 34/43), seizures (86%, 37/43), macrocephaly (the median percentile of head circumference is 90%), and paroxysmal deterioration (27%, 13/43). All the 43 patients' brain MRI satisfied typical MRI features proposed by van der Knaap. According to the analysis of the long-term follow-up, patients with type Ⅰ AxD began to have obvious regression in motor function after 7 years of age, and the social life ability was milally impaired 8(6, 10)scores at the last follow-up. Seventeen heterozygous missense mutations of GFAP were identified in 43 genetically confirmed patients, and 4 mutations were novel. The mutations in 41 patients (95%, 41/43) were de novo. Three hot spots of mutation in Chinese patients were found: p. Arg239(35%, 15/34), p. Arg79 (26%, 11/43) and p. R88 (16%, 7/43). Conclusion: The characteristic clinical manifestations of type Ⅰ AxD patients are developmental delay, seizures, macrocephaly and paroxysmal deterioration. Moreover, a few patients may present with brain stem symptoms, mental abnormalities, scoliosis or kyphosis. Patients with type Ⅰ AxD may show significant regression in motor function after 7 years of age.

Objective: To observe the effect of electroacupuncture(EA) combined with medication on changes of expression of Nestin, glial fibrillary acidic protein (GFAP) and neuron specific enolase (NSE) in the hippocampal CA 1 and CA 3 regions of focal cerebral ischemia (FC1) rats, so as to analyze its mechanisms underlying neuroprotection., Methods: Fifty male SD rats were randomly divided into normal control, model, EA, medication, and EA+ medication groups (n = 10 in each group). The FCI model was established by middle cerebral artery occlusion (MCAO) with thread embolus. EA (2 Hz, 2 V) was applied to the left "Hegu"(LI 4) and "Quchi" (LI 11) for 30 min, once daily for 14 days after MCAO. Rats of the medication group were given with intraperitoneal injection of gastrodin (10 mg/kg). The expression of Nestin, GFAP and NSE in the hippocampal CA 1 and CA 3 regions were detected by immunohistochemistry., Results: Compared with the normal control group, the numbers of Nestin- and GFAP-immunoreaction (IR) positive cells in both CA 1 and CA 3 regions of the hippocampus were significantly increased in the model ciroup (P<0.05), while those of NSE-IR positive cells in both CA 1 and CA 3 regions were significantly decreased in the mdlgroup (P<0.05). After EA and medication interventions, the numbers of Nestin- and NSE-IR positive cells in the CA 1 and CA 3 regions were evidently increased and GFAP-IR positive neurons were considerably reduced in the EA, medication and EA+ medication groups (P<0.05). The effects of EA+ medication were significantly superior to those of both EA and simple medication in up-regulating the number of Nestin- and NSE-IR positive cells and down-regulating the number of GFAP positive neurons in CA 1 and CA 3-regions (P<0.05)., Conclusion: EA and EA intervention combined with gastrodin can significantly up-regulate the number of Nestin- and NSE-IR positive cells, and down-regulate the number of GFAP positive cells in the CA 1 and CA 3 regions of hippocampus in focal cerebral ischemia rats, which may contribute to their effects in promoting the differentiation and proliferation of mature neurons in the hippocampus for improving cerebral functions. The effects of EA+ medication are obviously better than simple EA intervention.

Objective: To observe the effect of electroacupuncture (EA) preconditioning on the expression of neuronal nitric oxide synthase (nNOS), inducible nitric synthase (iNOS) and glial fibrilliary acidic protein (GFAP) in the cortex of focal cerebral ischemia-reperfusion (CI/R) rats so as to explore its underlying mechanism in the protection of ischemic cerebral tissue., Methods: Twenty-four Sprague-Dawley rats were randomized into sham operation (sham), model, and EA preconditioning groups (n = 8 in each group). The CI/R model was induced by intraluminal middle cerebral artery occlusion .(MCAO) with a nylon monofilament suture. Before modeling, EA (2 Hz/15 Hz, 3 V) was applied to "Baihui"(GV 20) and "Dazhui"(GV 14) for 30 min, once daily for 7 consecutive days. The neurologic impairment score was assessed by using Longa standards and the survival number of neurons in the local ischemic cerebral cortex was determined after Nissl staining, and the expression of nNOS, iNOS and GFAP in the cerebral cortex was detected using immunohistochemistry., Results: Compared with the sham operation group, the neurological deficit score of the rats in the model group was significantly increased (P < 0.01), and the number of survival neurons of the ischemic cortex was obviously decreased (P < 0.01), and the expression levels of nNOS, iNOS and GFAP were significantly increased in the model group (P < 0.01). In the EA preconditioning group , the neurological deficit score, the expression levels of nNOS and iNOS were significantly down-regulated (P < 0.01), while the number of the survival neurons and GFAP expression level in the ischemic cerebral cortex were obviously higher in the EA preconditioning group in compared with the model group (P < 0.01)., Conclusion: EA preconditioning can protect the ischemic cerebral cortex tissue from injury in CI/R rats, which may be related to its effects in down-regulating the expression of nNOS and iNOS, and up-regulating the expression of GFAP.

Objective: To observe the effect of electroacupuncture (EA) on the expression of proliferating cell nuclear antigen (PCNA) and glial fibrillary acidic protein (GFAP) in the subventricular zone (SVZ) of Parkinson's disease (PD) rats, so as to explore its mechanisms underlying improvement of PD., Methods: Eighteen SD rats were randomized into normal control, model and EA groups (6 rats/group). PD model was duplicated by microinjection of 6-hydroxydopamine (2.5 microg/microL, 6 microL) containing 0.02% antiscorbic acid into the right medial forebrain bundle region (P: -4. 0, L: 0.8, H: 8.0). On the 7th day after modeling, EA (100 Hz) was applied to bilateral "Hegu" (LI 4) and "Taichong" (LR 3) for 30 min, once a day for 21 days. The expression levels of tyrosine hydroxylase (TH), GFAP and PCNA in the right SVZ tissue were detected by immunohistochemistry., Results: TH expression was strong in the normal control group, but no TH expression was found in the model and EA groups. In comparison with the normal control group, the expression levels of PCNA and GFAP proteins and the number of PCNA-IR-positive cells in the right SVZ were decreased significantly in the model group (P < 0.01); while compared with the model group, the expression levels of PCNA and GFAP proteins and the number of PCNA-IR-positive cells in the right SVZ were up-regulated considerably in the EA group (P < 0.01, P < 0.05)., Conclusion: EA can up-regulate the expression of PCNA and GFAP proteins in the SVZ in PD rats without changes of TH expression, which may contribute to its effect in improving clinical symptoms of PD.

Objective: To delineate the phenotype and genotype characteristics in 12 Chinese children with Alexander disease (AD), which is helpful for the molecular diagnosis and genetic counseling in China., Methods: Clinical diagnosis of AD was based on MRI criteria proposed by van der Knaarp in 2001. Included AD patients were followed up for 0.50 - 3.67 years. Mutations in GFAP were detected by DNA sequencing., Results: The 12 cases of AD were clinically diagnosed. Age of first visit was 4.87 years (0.75 - 12.00 years), with 3 types of chief complaints: developmental delay in 3, recurrent seizures in 7, unable to walk after falling in 2. Average head circumference was 52.34 cm (44 - 58 cm), which larger than age-matched average by 6.45% (1.80% - 13.95%). On the first visit, scaling according to Gross motor functional classification system (GMFCS) was performed, with GMFCSI in 8, II in 3, V in 1. Mild to severe cognitive dysfunction were found in 8, and seizures in 11 cases. The 12 patients were followed up for 0.50 - 3.67 years, their motor and cognitive function remained stable. Episodic aggravations provoked by fever or falling were observed in 5 cases (41.67%). Heterozygous missense mutations of GFAP were detected in 12 patients. All mutations were de novo; 3 out of 10 mutations identified were novel. R79 and R239 were hot mutations, which was consistent with previous reports. Mutations were located in exon 1 in 8 cases., Conclusions: The phenotype in these patients is characterized by slower progression compared with reports from other population and high incidence of seizures. And episodic aggravations provoked by fever or falling were more common. The genotype characteristics are consistent with previous reports. The results of this research expanded the number of patients with Alexander disease found to have GFAP coding mutations in China.

Objective: To observe the dose-effect relationship of electroacupuncture of different current intensities combined with Morphine of different dosage on alleviating the rats' tibial cancer pain, and explore the possible mechanism, which could provide the experiment basis for alleviating the tibial cancer pain by electroacupuncture combined with Morphine., Methods: One hundred female Wistar rats were randomly divided into a normal group, a model group and eight treatment groups, 10 cases in each group. The rats in the treatment groups were treated by combined therapies of electroacupuncture of different intensities with 2 Hz /100 Hz dense-disperse wave on "Jiaji"(EX-B 2)and different dosage Morphine in 2 factor 3 level conditions, once a day for 6 days. The pain thresholds were observed before the treatment and 0 min, 1 h, 2 h and 5 h after the first treatment as well as after 3 and 6 times of treatments. The glial fibrillary acidic protein (GFAP) expression was determined by immunohistochemical method., Results: The rats' pain thresholds were significantly increased with electroacupuncture of 2 mA and 1 mA (all P < 0.01) on the 0 min, 1 h and 2 h of the first treatment, between which there were no significant differences (all P > 0.05). The pain threshold was still increased by electroacupuncture of 2 mA on the 5 h of the treatment (P < 0.01), while that of 1 mA failed to take effect (P > 0.05). After 3 and 6 times of treatments, both electroacupuncture of 2 mA and 1 mA had the effect of increasing the pain threshold (all P < 0.01), and the effect of 2 mA was superior to that of 1 mA (P < 0.05), had the synergistic effect with 5 mg/(kg x d) Morphine (P < 0.05). After 6 times of treatments, both electroacupuncture of 2 mA and 1 mA could inhibit the expression of GFAP (both P < 0.01), and there was no significant difference between them (P > 0.05). Both of 5 mg/(kg x d) and 2.5 mg/(kg x d) of Morphine, however, didn't bring about inhibition effect (P > 0.05)., Conclusion: There is a does-effect relationship on electroacupuncture of different current intensity for alleviating the tibial cancer pain in rats. The electroacupuncture with 2 mA, which is better than that with 1 mA, has the synergistic effect with 5 mg/(kg x d) of Morphine. The electroacupuncture can inhibit the expression of GFAP to cooperate with Morphine for the purpose of alleviating the rats' tibial cancer pain.

Objective: To study the effects of edaravone on glial fibrillary acidic protein (GFAP) and interleukin-1β (IL-lβ) expression and neuronal apoptosis in the juvenile rat hippocampus after status convulsion (SC)., Methods: One hundred and ninety-five juvenile male Sprague-Dawley (SD) rats were randomly divided into 3 groups: normal saline control and SC with and without edaravone treatment. Each of the 3 groups was further subdivided into subgroups sacrificed at 4, 12, 24, 48 and 72 hrs after SC (n=15). The SC model was prepared using lithium-pilocarpine. The expression of GFAP and IL-lβ protein was detected with immunohistochemistry methods. The neuronal apoptosis was observed by TdT-mediated dUTP nick end labeling (TUNEL). The hippocampal GFAP mRNA expression was detected by RT-PCR., Results: The value of IOD of GFAP and IL-lβ positive cells measured by immunohistochemistry in the untreated SC group increased compared with the control group. Expression of GFAP and IL-lβ protein was significantly reduced in the edaravone treated SC group compared with the untreated SC group. RT-PCR showed the expression trend of GFAP mRNA was similar to that of protein. The TUNEL positive cells in the hippocampus CA1 in the untreated SC group increased significantly 12 hrs after SC and reached a peak at 48 hrs compared with the control group. The intervention with edaravone decreased significantly TUNEL positive cells between 12-48 hrs after SC, but the number of TUNEL positive cells in the intervention group remained significantly greater than in the control group., Conclusions: The expression of GFAP and IL-lβ in the hippocampus increases after SC in rats. Edaravone may decrease the expression of GFAP and IL-1β and reduce the number of neuronal apoptosis. These results suggest that edaravone may have protective effects against brain damage caused by SC.

Objective: To explore the changes in the expressions of glial fibrillary acidic protein (GFAP) and growth- associated protein-43 (GAP-43) in retinal ganglial cells after neural transplantation., Methods: Thirty-nine rats were randomized into normal control group, nerve amputation group and nerve amputation with peripheral nerve transplantation group. Immunohistochemistry was used to detect the changes in the expressions of GFAP and GAP-43 at different time points after the operations, and real-time PCR was employed to detect the mRNA expressions of 13 genes in the retinal ganglial cells of the rats., Results: Immunohistochemistry showed obviously increased GFAP expressions in the retina following the nerve amputation. GFAP expression was down-regulated while GAP-43 expression upregulated in the retinal ganglial cells after peripheral nerve transplantation. Real-time PCR results showed that 5 days after the operations, retinal GFAP and GAP-43 expressions increased significantly in the nerve amputation group and peripheral nerve transplantation groups as compared with those in the control group, but GAP-43 expression decreased significantly in the former two groups afterwards., Conclusion: The regenerated retina may adjust the production of GFAP. The retinal ganglial cells express GAP-43 during retinal regeneration. Up-regulation of the expression of GAP-43 provides the evidence for nerve regeneration following the nerve transplantation.

Objective: To observe the effect of electroacupuncture (EA) on the expression of spinal glial fibrillary acidic protein (GFAP), tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) in chronic constriction injury (CCI) rats., Methods: Seventy-two SD rats were randomized into sham operation (sham), CCI (model) and EA groups (n = 24/group). The mechanical and thermal pain thresholds were measured by using Von Frey filaments and radiant-heat irridiation separately. The immunoactivity of GFAP of spinal dorsal horn (L4-L5) was assessed by immunohistochemistry, and the expression of spinal TNF-alpha mRNA and IL-1beta mRNA was detected by real time-PCR., Results: Compared with pre-CCI and sham group, both mechanical and thermal pain thresholds decreased considerably in rats with CCI (P < 0.05), and in comparison with model group, those of EA group increased markedly (P < 0.05). Compared with sham group, GFAP immunoactivity (mainly in the lamina I-II of the spinal dorsal horn), TNF-alpha mRNA and IL-1beta mRNA expression in the ipsilateral spinal cord on the CCI side in model group increased considerably (P < 0.05), while compared with model group, the expression of GFAP, TNF-alpha mRNA and IL-1beta mRNA in EA group was down-regulated remarkably (P < 0.05)., Conclusion: EA can effectively suppress CCI-induced up-regulation of expression of spinal GFAP, TNF-alpha mRNA and IL-1beta mRNA, which may contribute to its effect in reducing mechanical allodynia and thermal hyperalgesia in neuropathic pain rats.

Objective: To determine the expression of glial fibrillary acidic protein(GFAP) and vascular endothelial growth factor(VEGF) in the hippocampus of rats with Alzheimer's disease(AD), and to determine the effect of butylphthalide on them and its significance., Methods: Sixty male adult rats were randomly divided into a model group, a Butylphthalide group, and a control group. AD models were established by injecting beta-amyloid protein 1-42 into the hippocampus of rats. Sixty days later,the rats were sacrificed and both sides of the hippocampus were sectioned for immunohistochemistry., Results: Positive cells of GFAP in the hippocampus of the model group increased and the expression of VEGF decreased statistically, compared with the control group(P<0.01). The positive cells of GFAP in the hippocampus of the butylphthalide group decreased and the expression of VEGF increased significantly, compared with the model group(P<0.05)., Conclusion: Butylphthalide may protect the neuron-vascular unit of the hippocampus of Alzheimer model rats by inhibiting the expression of GFAP and increasing the expression of VEGF.

Objective: To observe the effects of auricular acupuncture on the learning and memory abilities of model rats with Alzheimer's disease (AD), and investigate its mechanism., Methods: Thirty SD rats were randomly divided into a control group, a model group and an auricular acupuncture group, 10 rats in each group. The model rats with AD were established by multiple injections with Okadaic Acid into the CA1 region of hippocampus. In the control group, the same quantity injection with Dimethyl Sulfoxide (DMSO) was applied on experimental rats. The auricular acupoints of "Nao" (brain) and "Shen" (kidney) were used for treating in the auricular acupuncture group, in contrast, the auricular region were not treated in the model and the control groups. The learning and memory capabilities of the rats were assessed with Morris Water Maze behavioral test, and the expressions of choline acetyltransferase (ChAT) and glial fibrillary acidic protein (GFAP) were examined by immunohistochemistry., Results: Comparing with the model group, the treated AD rats with auricular acupuncture was showed that the average escape latency was obviously shortened in the place navigation test (P<0.01), the movement time in plateform quadrant was obviously prolonged in the spatial probe test (P<0.05), and the number of traversing platform obviously increased (P<0.01) after the platform was taken away. The expression of ChAT increased in the hippocampus and cortex (P<0.01, P<0.05), but the expression of GFAP obviously decreased in the CA1 region of hippocampus (P<0.01)., Conclusion: Auricular acupuncture can improve the learning and memory capability of the model rats with AD. Its mechanism might be related with decreasing cholinergic neuron damage and reducing the abnormal activation and hyperplasia of astrocyte.

Objective: To explore the influence of hyperbaric oxygen (HBO) treatment on neural plasticity and it's mechanism in experimental rats with cerebral ischemia., Methods: Ninety-healthy male adult Sprague-Dawley rats (3 to approximately 4 month old) were randomly divided into a pseudo-operative group, a model group, and an HBO therapy group. The middle cerebral artery occlusion model was duplicated with suture methods, then we used beam walking test (BWT) to determine the motor skill of the rats and immunohistochemistry method to detect the distribution and location of microtubule-associated protein-2 (MAP-2) and glial fibrillary acidic protein (GFAP). Quantitative real-time PCR was used to detect the expression of Map-2 mRNA and GFAP mRNA., Results: Immunohistochemistry showed that the fluorescence gray scale value of Map-2 in the HBO group was the highest in 3 groups at 1st week and 2nd week (P<0.05).The value of GFAP was lower than that of the model group but higher than that of the sham operated group (P<0.05). Real-time fluorescence-quantitative PCR indicated that the Map-2 mRNA of HBO group was the highest in 3 groups at 1st week and 2nd week (P<0.05); but the value of GFAP mRNA in the HBO group is lower than that of the model group,but higher than that of the sham operated group at 1st week and 2nd week (P<0.05)., Conclusion: After cerebral infarction, giving hyperbaric oxygenation treatment can improve the limbs motor function, and hyperbaric oxygenation treatment can increase the expression of Map-2 and decrease the expression of GFAP, which promote neural plasticity.

Objective: To observe the expressions of nestin and glial fibrillary acidic protein (GFAP) and their association with reactive astrocytes following spinal cord injury in adult rats., Methods: Adult rats with compression injury of the spinal cord were divided into 7 groups (n=6) and examined at 1, 3, and 5 days and at 1, 2, 4 and 8 weeks after the injury. The recovery of the locomotor function after the injury was evaluated with Basso, Beattie and Bresnahan (BBB) scale, and the degree and scope of the spinal injury were assessed using toluidine blue staining. Immunohistochemistry, double immunofluorescent labeling and an image analysis system were employed to observe nestin and GFAP expression and cell proliferation in different regions of the spinal cord., Results: The bilateral hind limb locomotor function of the rats declined severely 24 h after the spinal cord injury and underwent substantial recovery in 1 or 2 weeks after the injury, but followed by rather slow recovery afterwards. Toluidine blue staining of the spinal cord 24 h after the injury showed significant pathological changes in the neurons. The extension of the tissue injury increased with time till 1 week after the spinal cord injury. The site of injury and the adjacent tissues presented with markedly increased nestin and GFAP expressions 24 h after the injury, and nestin+/GFAP(-) cells dominated in the ependymal region around the central canal, whereas nestin+/GFAP+ dominated in the in other regions, showing significant difference from the control group. Nestin and GFAP expression reached the peak level 3 to 7 days after the injury and declined gradually till reaching nearly the control level at 2 weeks., Conclusion: Compression injury of the spinal cord induces up-regulated expressions of nestin and GFAP, and nestin expression is positively correlated to the reactive astrocytes, which, along with the neural stem cells, respond to spinal nerve injury and possibly play a role in repair of the central nervous system injury.

Objective: To study clinicopathologic features, treatment and prognosis of pilocytic astrocytoma (PA)., Methods: Histopathological, ultrastructural, immunohistochemical (EnVision method) and clinical features of 68 cases of PA were studied by microscopic investigation with correlation of clinical follow-up information when available., Results: Thirty-five male patients and 33 female patients were studied. The patient's age ranged from 3 to 66 years (mean = 20.1 years). The mean time from symptom onset to surgery was 371 days (range, 3 days to 14 years). Cystic degeneration was noted in 41 cases (60.3%), and enhancement of the tumor was noted in 43 cases (87.8%). On postcontrast imaging examination there were 33 cases involving the cerebellum (48.5%). Total tumor excision was performed in 35 patients, subtotal tumor excision was performed in 31 patients, and the procedures of other 2 patients were not clear. Among 51 patients with follow-up information, 44 were alive, 7 had recurrent tumor, and 7 died. The post-operative survival ranged from 2 months to 124 months (mean survival = 48.1 months). Five years and ten years survival rates were 89%, respectively. Tumors with classic histopathology demonstrated biphasic pattern of growth, consisting of compact elongated bipolar astrocytes associated with rosenthal fibers, and less cellular areas of multipolar cells with granular bodies and microcyst. Some cases showed atypia of nuclei, and occasional mitoses. Involvement of subarachnoid space was seen in 17 cases. One case had anaplastic features. All cases showed diffuse positive staining for GFAP and low expression for Ki-67, except 1 anaplastic tumor with 10% Ki-67 indices. Tumors with subarachnoid space involvement showed positive reticular fiber staining and negative EMA staining., Conclusions: PA is a benign, WHO grade I tumor with favorable prognosis, and does not require radiotherapy after total resection. The tumor can be mistaken as higher-grade astrocytoma when involving the subarachnoid space, and with cytological atypia, leading to unnecessary radiotherapy after surgery. Recurrence rate is increased when only partial resection is achieved. The outcome for patients with brainstem tumor or anaplastic PA is poor.

Objective: To investigate whether gliosis in the brain tissues of the hamsters infected with various amounts of scrapie strain 263K is correlated with the inoculation doses or the incubation times., Methods: The total values of glial fibrillary acidic protein (GFAP) in brains were evaluated by Western Blots and the GFAP-stained cells were detected by immunohistochemistry (IHC). The characteristics of GFAP distributions among various groups were defined by quantitive and statistic analyses., Results: Compared with the brain tissues of normal hamsters, remarkably higher total GFAP levels and more GFAP-stained cells were observed in the brain tissues of infected ones, howbeit, no significant difference was addressed among the infected groups., Conclusion: Inoculations of various amounts of scrapie strain 263K into experimental hamsters intracerebrally induced the similar patterns of gliosis in the brains at the clinically terminal stage, regardless of infectious doses and incubation times.

Objective: To investigate the expression and significance of peroxisome proliferators-activated receptor gamma (PPAR gamma) in human glioma., Methods: Immunohistochemical staining for PPAR gamma was performed using biopsy specimens of human glioma of various histological types. Expression of PPAR gamma and GFAP in glioma cell lines SWO-38, U251 and SHG-44 were analyzed using Western blotting and reverse transcription-polymerase chain reaction (RT-PCR)., Results: Immunohistochemical study showed that PPAR gamma was expressed in glioma tissues with positive rate of 37.5%. Western blotting and RT-PCR showed that PPAR gamma was expressed in both glioma cell lines SWO-38 and U251, but not in SHG-44 cells. However, high expression of GFAP was detected in SHG-44 cells., Conclusion: PPAR gamma is associated with carcinogens of glioma. Actived PPAR gamma by agonist may be a novel approach to the treatment of glioma.

Objective: To study the potential interaction between PrP protein and glial fibrillary acidic protein (GFAP) and identify the binding region within PrP with GFAP., Methods: The supernatant of healthy and scrapie-infected hamsters' brain homogenate was prepared, while various recombinant PrP or GFAP proteins were expressed using prokaryotic-expressing or in-vitro translation system. The possible molecular interaction between PrP proteins and GFAP was tested by Pull-down and immunoprecipitation assays., Results: Both native PrP(C) and its protease-resistant isoform (PrP(Sc)) formed complexes with the native GFAP. The full-length recombinant PrP proteins interacted with GFAP. The domain responsible for interacting GFAP was located at C-terminal of PrP (residues 91 to 231)., Conclusion: The studies of the association of PrP with GFAP may further provide insight into a potential role of GFAP in the biological function of PrP and the pathogenesis of prion disease.

Objective: To investigate the feasibility of inducing adult human myoblasts into neural precursor cells., Methods: The myoblasts were isolated with mixed digestive enzyme from minced human temporal muscle samples, cultured and purified clonally. The 3rd passage cells were incubated with serum free medium including basic fibroblast growth factor (bFGF), epidermal growth factor (EGF) and leukemia inhibitory factor (LIF). Morphological change was investigated during incubation period. Immunofluorescence cytochemistry and RT-PCR analysis were used to assess cell differentiation and trans differentiation., Results: After the induction, cells became non-adherent aggregates as neurospheres. The myoblast-derived neurospheres was immuno-positive for nestin. In differentiation condition, they looked like neurons and glial cells and expressed neuronal (microtubule associated protein 2, MAP-2), astrocytic (Glial fibrillary acidic protein, GFAP) and oligodendrocytic (Galactocerebroside, Galc) markers by immunocytochemistry. The result by RT-PCR was coincident with immunocytochemistry. The myoblast-derived neurospheres expressed MAP-2 and GFAP after they were transplanted into the brain of rats with cerebral ischemia., Conclusion: Adult human myoblasts can be inducted to trans-differentiate into neural precursor cells.

Objective: To study the effect of saikosaponins, the active ingredients of Bupleurum chinense DC, on glial fibrillary acidic protein (GFAP) expression in hippocampal astrocytes of chronic kindling rats induced by pentetrazole (PTZ)., Methods: Forty-eight healthy Sprague-Dawley rats were randomized into 6 equal groups, namely the blank control group (Group A), normal saline group (Group B), sodium valproate group (Group C), and 3 saikosaponins groups of high, medium and small doses (Groups D, E, and F, respectively). The rats (except those in Group A) received intraperitoneal injection of PTZ to induce chronic kindling 1 h after the respective agents as indicated were administered intragastrically on a daily basis for 4 consecutive weeks. Upon completion of the treatment course, the rats were sacrificed and the brain tissues were sampled, sliced and stained for immunohistochemical examination. The results were analyzed to calculate the positive cell count, cross-sectional area of the cells and the gray scale., Results: In group B, the positive cell population and cross-sectional area of the positive cells were the greatest among the groups (P<0.01), but the positive cell gray scale of the CA1 and CA2 regions and the dentate gyrus (DG) of the hippocampus was the lowest. The CA1 region of Group B was significantly different from that of groups A, C and D (P<0.01), and the CA2 region different from groups A, C, D and E (P<0.05), while the DG different from group F (P<0.05) and groups A, C, D and E (P<0.01)., Conclusion: In chronic kindling rats induced by PTZ, GFAP overexpression can be inhibited by saikosaponins, which suppress the abnormal activation of hippocampal astrocyte of the kindling rats.

Purpose: To modulate optic nerve regeneration by reducing the inhibitory effects of reactive astrocytes., Methods: Bcl-2 over-expression transgenic mice were mated with GFAP/Vimentin double gene knock out mice (GFAP-/-/Vim-/-) to produce Bcl-2 tg/GFAP-/-/Vim-/- triple mutant mice (3M). Optic nerve crush model was established respectively in 3 P5 and 3 P18 mice. GAP43 immunofluore-scence was used to specifically label regenerative axons., Results: Optic nerve regeneration was observed in P5 3M mice all the way to optic chiasm in P18 3M mice, only local sprouting regeneration was observed., Conclusions: With reestablishment of intrinsic ability of retinal ganglion cells by Bcl-2 transgenic, optic nerve regeneration is promoted by reducing the inhibitory effects of reactive astrocytes, which demonstrates reactive astrocyte as an important factor inhibiting optic regeneration and provides new direction for regenerative therapy in glaucoma.

Objective: To isolate and identify human retinal stem cells (HRCs) from human embryonic retina., Methods: HRCs were isolated from neural retinas at the 16th-20th week of gestation and cultured in either serum-free or serum-containing media. The expression of nestin antigen and retinal terminal cell markers were conformed by immunofluorescence labeling technique. RT-PCR analysis was used to identify the difference of nestin gene expression in HRCs and differentiated cells., Results: HRCs divided into multiple generations (up to passage 8) and expressed nestin. Other characteristics of the isolated cells were to differentiate into glial fibrillary acidic protein-positive glial cells (Müller cells) and retina-specific neurons expressing rhodopsin, syntaxin, PKCalpha, and Thy1. These were markers for rod receptors, amacrines, bipolar cells, and ganglion cells respectively. When cultured with serum, the expression of nestin was significantly downregulated in HRCs., Conclusion: Human neural retina at the 16th-20th week of gestation harbors neural stem cells that are capable of selfrenewal and multilineage differentiation.

Objective: To study the genes change of hMSC before and after differentiation into neuron-like cells., Methods: hMSC were separated from marrow, cultured and expanded in culture medium. After hMSC being induced to differentiate with Shenqiye, Neuron-specific enolase (MSE), neurofilament (NF), and glial fibrillary acidic protein (GFAP) were detected by immunocytochemistry respectively. Using semi-quantitiative RT-PCR to detect ten genes change in hMSC before and after differentiation. Two genes were used to detect the gene sequence., Results: When treated with the inducer--Shenqiye, hMSC exhibited neuronal phenotype. The expressions of NSE and NF in the neuron-like cells were positive, but the glial astrocyte marker GFAP was not found. hMSC expressed germline, ectodermal, endodermal and mesodermal genes prior to neurogenasis. After differentiation, germline genes did not express, while the expressions of endodermal and mesodermal genes were weakened. Ectodermal and neuronal genes expressed stronger., Conclusion: hMSC express multipotentiality, and the neural differentiation comprises quantitative modulation of gene expression rather than simple on-off switching of neural specific genes.

Objective: To investigate the molecular basis of infantile Alexander disease in a Chinese patient, which may yield useful information for further genetic counseling., Methods: DNA sequencing analysis and restriction endonuclease analysis were used to detect the mutation of glial fibrillary acidic protein (GFAP) gene in a patient with clinically diagnosed Alexander disease, in her parents and in 50 healthy controls., Results: A 249C>T (R79C) mutation was identified in the exon 1 of the GFAP gene but not in her parents and the controls., Conclusion: The study on mutation of GFAP gene in Chinese patients with Alexander disease has never been reported previously. The mutation analysis of GFAP gene can provide valuable information for the diagnosis of Alexander disease and can serve as a reliable method of prenatal diagnosis for the family.

For specific expressing Cre recombinase in central nerve system (CNS), a transgenic construct (pGFAP-Cre-hGH), containing the beta-globin insulators, 1.8 kb of glial fibrillary acidic protein gene (GFAP) 5' end regulation region, Cre gene and polyA of human growth hormone gene (hGH) was generated, in which the 5' end regulation region of GFAP was isolated from a 129sv mouse genomic DNA library with PCR-screening. 7.6 kb of pGFAP-Cre-hGH DNA fragment was introduced into 191 fertilized eggs by microinjection. 176 injected eggs were implanted into the oviducts of eight female mice respectively, from which 25 offspring were obtained. Seven mice carry the Cre genes by the identification of PCR and Southern blotting, and the integration efficiency is 28%. The GFAP-Cre transgenic mice were crossed with ROSA26 mice whose genomic DNA is integrated by LoxP sites and LacZ expression frame to check the activity and the tissue-specific expression of Cre recombinase and recombination with its mediation between two LoxP sites. The results of LacZ dying showed that the Cre recombinase was expressed only in CNS and successfully mediated the recombination between the LoxP sites in vivo.

Objective: To investigate the biological effects of ectopic overexpression of glial fibrillary acidic protein (GFAP) in human malignant glioma cell line, and to explore new method of differentiation induction gene therapy for gliomas., Methods: A eukaryotic expression vector containing 1.1 kb GFAP cDNA fused with green fluorescent protein (GFP) gene, pIRGFP-GFAP, was transfected into human SHG-44 glioma cell line by lipofectamine. The expression of GFAP/GFP gene and their proteins were detected by fluorescent real-time monitoring, in situ hybridization, Western blot and immunocytochemistry. Flow cytometry, soft agar colony formation and other methods were used to measure the effects of exogenous GFAP expression on cell cycle progression, morphology and growth features of the transfected glioma cells., Results: The expressions of GFAP mRNA and its protein were markedly increased in SHG-44 cells upon stable transfection with pIRGFP/GFAP vector. Profound morphological changes in these cells were also observed, including the formation of abundant, stellate and thin cytoplasmic processes and a reduction of atypia. Cell proliferation rate and its tumorigenecity on soft agar were markedly reduced. In addition, cell cycle analysis revealed a percentage decrease of cell populations at G0/G1 and G2/M phases., Conclusions: Ectopic overexpression of GFAP gene could significantly suppress the growth of SHG-44 malignant glioma cells along with an induction of differentiation. These results imply that forced over-expression of GFAP gene may provide a new strategy for glioma therapy.

Objective: In order to explore the effects of lead on the growth and development of cultured hippocampal neural cells and on the expression of Oct-2, the II subtype POU domain protein., Methods: Experiment cell model was established using primary culture of hippocampal neural cells from SD rat embryos. Target cells were exposed to lead acetate in the different concentrations, i.e. 10(-1), 10(0), 10(1), 10(2), 10(3) micromol/L, while the control group was given the same quantity of the culture medium. The immunohistochemistry method was utilized to detect the expressions of Neurofilament (NF) and Glial Fibrillary Acidic Protein (GFAP), the markers for neuron and astrocyte, respectively, and the expression of Oct-2 as well., Results: The results showed that 10 micromol/L lead acetate treatment caused diminishing of neuronal cell body and the decreases of both axon lengths and inter-cellular connections. In addition, 1 micromol/L lead acetate significantly increased the number of GFAP-positive cells compared with the control group (P < 0.05). By image analysis system, 1 micromol/L lead acetate treatment was found to induce a statistically significant increase of the positive area rate concerning Oct-2 expression in hippocampal neurons and astrocytes, while both positive area rate and integral density of light of Oct-2 expression were found to increase markedly in the groups treated by 10 micromol/L lead acetate (P < 0.01)., Conclusions: Lead acetate treatment may contribute to the inhibitions of both growth and differentiation of hippocampus neurons, and to the stimulation of glial cell hyperplasia simultaneously. In addition, the CNS impairments caused by lead is partly correlated with the enhancement of Oct-2 expression.

Objective: To investigate the expression of glial fibrillary acidic protein (GFAP), GFAP mRNA and interleukin-1beta mRNA (IL-1beta mRNA), tumor necrosis factor-alpha mRNA (TNF-alpha mRNA) in neonatal rat brain after intrauterine infection., Methods: Escherichia coli (E. coli) was inoculated into both uterine horns of pregnant rats when gestation was 70% complete (15 days). The control group was treated with normal saline. The pups were killed on the postnatal day 1 (P1), P3 and P7, respectively. The cerebral white matter damage of the neonatal rats was determined by HE staining. Immunohistochemistry was used for evaluation of GFAP expression in neonatal rat brains and RT-PCR to analyze GFAP mRNA, IL-1beta mRNA and TNF-alpha mRNA expression at P1, P3 and P7., Results: The major histopathological changes in neonatal cerebral white matter at P7 after intrauterine infections were: weak staining of cerebral white matter and focal rarefaction. GFAP-positive cells were observed in both the control and the E. coli-treated groups. The numbers of GFAP-positive cells of the E. coli-treated group pups were markedly increased in periventricular white matter and hippocampus at P7 compared with those of the control group (periventricular white matter: 9.73 +/- 3.55 vs 5.67 +/- 1.90, P < 0.05 and hippocampus: 7.81 +/- 3.61 vs 2.16 +/- 1.11, P < 0.05, respectively). No significantly different levels of GFAP expression in corpus callosum were found between two groups (P > 0.05). The expression of GFAP mRNA in brain of the E. coli-treated neonatal rat was higher than the control at P1, P3 (P1: 0.25 +/- 0.07 vs 0.15 +/- 0.08, P < 0.05 and P3: 0.50 +/- 0.09 vs 0.39 +/- 0.08, P < 0.05, respectively), but the expression of GFAP mRNA in brain of the neonatal rat at P7 had no significant difference between two groups (P > 0.05). The expression of IL-1beta mRNA and TNF-alpha mRNA in brain of the E. coli-treated neonatal rat were higher than of the control at P1 (IL-1beta mRNA: 0.83 +/- 0.19 vs 0.50 +/- 0.30, P < 0.05 and TNF-alpha mRNA: 0.74 +/- 0.30 vs 0.30 +/- 0.20, P < 0.05, respectively), but the expression of IL-1beta mRNA and TNF-alpha mRNA in brain of the neonatal rat at P3 and P7 had no significant difference between two groups (P > 0.05)., Conclusions: The intrauterine infection could cause neonatal white matter damage and IL-1beta, TNF-alpha may be a mechanism mediating between the two events.

Objective: To investigate the ability of human GFAP positive neural progenitor cell line from the subventricular zone (SVZ) to differentiate into neurons., Methods: Real-time RT-PCR, Western blot analysis and immunocytochemistry were used to examine the expression level of the neural stem cell marker and neuronal-specific marker before and after all-trans-retinoic acid (AT-RA) induction in the GFAP positive neural progenitor cell line. Immunocytochemistry was used to examine the expression of the neuronal-specific marker after transplantation the GFAP positive neural progenitor cell line into the animal model., Results: After induction, in the GFAP positive neural progenitor cell line the expression levels of the neuronal-specific marker increased, while the neural stem cell marker decreased both in mRNA and protein levels. After transplantation into animal model, the GFAP positive neural progenitor cell line could differentiate into neurons., Conclusion: The GFAP positive neural progenitor cell line could be induced to differentiate into neurons both in vitro and in vivo.

Primers for the glial fibrillary acidic protein (GFAP) gene were designed from a human cDNA sequence aligned with the mouse GFAP gene on the principle of comparative anchor tagged sequence (CATS). The 412 bp PCR product isolated from Chinese Erhualian pig genome was characterized as the porcine GFAP gene by comparing the sequence with the GenBank database. The chromosomal location of the GFAP gene is on pig Chr: 12(p11-(2/3)p13) using pig x rodent somatic cell hybrid panel.

The present study was to investigate the changes of macroglia and the effect of gonadal hormone on reactive gliosis and of antisense GFAP retrovirus on astrocyte, gliosis and glial scar formation after brain stab injury (BSI). The results indicate that astrocytes are the main cells of glial scar. GFAP plays an important role in the maintenance of structure and function of astrocytes. The oligodendrocyte is not an active cell during glial scar formation. The gonadal hormone can modify the reactivity of astrocyte after BSI, but has no significant effect on differentiation and proliferation of astrocyte. The recombined GFAP retrovirus can effectively inhibit the growth of astrocyte, and GFAP expression of injured astrocyte in vitro and glial scar forming in vivo.

Objective: To study the relationship between reactive astrogliosis and its autocrine bFGF., Methods: On the primary astrocyte culture with mechanical scratch, the dynamic expression of bFGF, PCNA, GFAP and GFAP-mRNA were determined by immunocytochemistry and in situ hybridization., Result: Astrocytes at the edge of the injury began to express bFGF 2 hours after scratching and reached its peak by the 12th hour, then declined after 2 days. GFAP-mRNA was detectable in astrocytes near the injury from the 6th hour and reached its peak within 24 hours. A few hypertrophic astrocytes in the injured area expressed GFAP-mRNA on the 3rd day. Enhanced GFAP expression of astrocytes was observed with hypertrophy of cytoplasma which extended wide processes into the injured area from the day of the scratch and reached its peak on the 2nd day. On the 3rd day, the previously injured area was covered with hypertrophic astrocytes. PCNA was expressed 2 hours after damage by some of the astrocytes in the vicinity of injury., Conclusion: Autocrine bFGF was the early response of reactive astrocytes and bFGF may act as promoter of reactive astrogliosis. Enhanced GFAP expression was the result of upregulation of GFAP-mRNA. The prominent event of reactive astrogliosis was the hypertrophy of astrocytes.