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Objective: This study aims to observe changes in the expression of the hepatic endoplasmic reticulum stress PERK-eIF2α-ATF4 signaling pathway protein in the progression of CCl 4 -induced hepatic fibrosis in rats., Methods: Male Wistar rats were sacrificed at the end of the 2 nd , 4 th , 8 th and 12 th weeks, respectively. A specific test was performed to compare the pathological changes of hepatic tissues in the model and normal groups. Immunohistochemical staining and Western blot were carried out to detect the expression of p-PERK, p-eIF2α and ATF4 proteins in the hepatic tissue group. Furthermore, real-time PCR was used to detect changes in the expression of ATF4 mRNA in hepatic tissues., Results: In the eight-week and twelve-week hepatic fibrosis group, significant fibrosis hyperplasia was identified in the livers of rats, and pseudo-lobules were also formed in the livers of rats in the twelve-week hepatic fibrosis group. Immunohistochemical staining and Western blot results indicated that the expression levels of p-PERK, p-eIF2α and the ATF4 protein in the livers of rats were significantly increased from the 8 th week compared with the normal group ( P <0.05). Real-time PCR results revealed that the expression of ATF4 mRNA was significantly increased in hepatic tissues in the hepatic fibrosis group compared with the normal group ( P <0.05), and this was gradually enhanced as hepatic fibrosis progressed., Conclusions: CCl 4 can induce an increase in the expression of the PERK-eIF2α-ATF4 signaling protein in the development of hepatic fibrosis along with phosphorylation-mediated activation, indicating that the activation of the PERK-eIF2α-ATF4 signaling pathway may contribute to the onset and development of hepatic fibrosis by regulating downstream target genes., Competing Interests: None., (IJCEP Copyright © 2018.)

Objective: To compare and analyze the differential expression of proteins between coal-burning arsenism serum and normal human serum and identify the proteins related with arseniasis caused by coal-burning., Methods: Serum samples were collected from 6 normal subjects and 6 patients suffering from coal-burning arsenism. 2-DE was performed to separate serum proteins. After silver staining, the differential expression of proteins was analyzed and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS)., Results: There were an average of 779 +/- 35 spots and 865 +/- 30 spots on 2-DE matching of two groups and the matching rate was 90.1% between two groups. From these two groups, 60 different protein spots were identified. Up-regulated expression was observed in 25 proteins and down-regulated expression in 35 proteins in the patient serum group. Among which 35 with differential expression above three times were singled out and MALDI-TOF-MS analysis was carried out on them. Thirteen proteins were identified, including keratin 10, apolipoprotein A-V, transferrin, alpha-1-antitrypsin, human zinc-alpha-2-glycoprotein, mitogen-activated protein kinase 3, vacuolar protein sorting 33A, O-linked GlcNAc transferase and etc. Up-regulated expression was observed in 5 proteins and down-regulated expression in 8 proteins in the patient serum group., Conclusion: The well-resolved and reproducible 2-DE serum patterns of patients suffering from coal-burning arsenism were established and some differentially expressed proteins characterized. These data will be used to screen the biomarker and to further study arseniasis caused by coal-burning.

Objective: To develop the 2-DE profiles of proteome from hepatic fibrosis tissue and preliminarily analyze the differential expression of proteome., Methods: The differential expression of proteins were analyzed by imaging analysis and MALDI-TOF-MS after the total protein was extracted from the hepatic fibrosis tissue and the normal liver tissue by 2-DE. 3 proteins were verified by in Western-blotting., Results: Fifty-nine differentially expressed protein were found in the proteome profile analysis of these two types of tissue, among which 30 protein spots were up-regulated and 29 protein spots were down-regulated in these hepatic fibrosis tissues. Fifteen protein spots with the differential expressions more than 3 times were analyzed by peptide mass fingerprint (PMF) and 10 differentially expressed proteins were found to be related with cell signal transduction, cell proliferation, and oxidative stress. Western-blotting results of 14-3-3beta, DJ-1, and PEBP were consistent with those by the 2-DE examination., Conclusion: Some differentially expressed proteins have been found in the hepatic fibrosis tissue and the normal liver tissue. These data provided a fundamental basis for further study of the mechanism of hepatic fibrogenesis. Proteome;

Aim: To investigate the effects of Danshao Huaxian (DSHX) capsules, a preparation of traditional Chinese medicine, on the expression of matrix metalloproteinase-1 (MMP-1), and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the fibrous livers of rats., Methods: Eighty male Wistar rats were randomly divided into normal control group (group A), CCl(4)-induced hepatic fibrosis group (group B), non-DSHX-treated group (group C), low dose-treated group (group D), and high dose-treated group (group E). Fibrous liver models in rats were induced by subcutaneous injection of CCl(4), oral administration of alcohol and high-lipid/low-protein diet for 8 wk. After the models were established, the rats in groups D and E were orally given a low dose (0.5 g/kg) and a high dose (1.0 g/kg) of DSHX daily for 8 wk, respectively. Then, the liver indexes, serum hyaluronic acid (HA) and alanine aminotransferase (ALT) were examined. The degree of hepatic fibrosis was evaluated by optical microscopy. Hydroxyproline (Hyp) in the urine was determined, and the expression of MMP-1 and TIMP-1 was detected by immunohistochemical techniques., Results: In groups D and E, the liver indexes, levels of serum HA and ALT reduced and development of hepatic fibrosis weakened significantly. The urinary Hyp and expression of MMP-1 in the liver tissues elevated, but the expression of TIMP-1 decreased obviously, as compared to groups B and C., Conclusion: DSHX enhances the expression of MMP-1 but decreases that of TIMP-1 in liver tissues of CCl(4)-induced hepatic fibrotic rats, which may result in its elevated activity that contributes to fighting against hepatic fibrosis.

Aim: To investigate the effects of Danshaohuaxian (DSHX), a Chinese herbal recipe, on the apoptosis and cell cycles of hepatic stellate cells (HSCs) in rat hepatic fibrosis and its possible mechanisms., Methods: Seventy-six male Wistar rats were randomly divided into normal control group, hepatic fibrosis group, non-DSHX-treated group and DSHX-treated group. Except for the normal control group, rat hepatic fibrotic models were induced by subcutaneous injection of carbon tetrachloride (CCl4), drinking alcohol, giving diet of hyperlipid and hypoprotein for 8 wk. When the hepatic fibrotic models were produced, 12 rats of hepatic fibrosis group (15 rats survived, others died during the 8 wk) were sacrificed to collect blood and livers. HSCs were isolated from the other 3 rats to detect the apoptotic index (AI) and cell cycles by flow cytometry. DSHX was then given to the DSHX-treated group (1.0 g/kg, PO, daily) for 8 wk. At the same time, normal control group and non-DSHX-treated group were given normal saline for 8 wk. At end of the experiment, some rats in these three groups were sacrificed to collect blood and livers, the other rats were used for HSC isolation to detect the apoptotic index (AI) and cell cycles. Then the liver index, serum hyaluronic acid (HA) and alanine aminotransferase (ALT), degree of hepatic fibrosis, urinary excretion of hydroxyproline (Hyp) and expression of collagen types I and III (COL I and III) in these four groups were detected respectively., Results: Compared with the indexes of the hepatic fibrosis group and non-DSHX-treated group, the DSHX-treated group revealed a liver index of (0.0267+/-0.0017 vs 0.0423+/-0.0044, 0.0295+/-0.0019, P<0.05), levels of serum HA (200.78+/-31.71 vs 316.17+/-78.48, 300.86+/-72.73, P<0.05) and ALT (93.13+/-5.79 vs 174.5+/-6.02, 104.75+/-6.54, P<0.01), and stage of hepatic fibrosis (1.30 vs 4.25, 2.60, P<0.01) all reduced. The urinary excretion of Hyp increased (541.09+/-73.39 vs 62.00+/-6.40, 182.44+/-30.83, P<0.01), the COL I and III expression decreased (COL I: 1.07+/-0.96 vs 4.18+/-2.26, 3.22+/-1.44, P<0.01; COL III: 1.09+/-0.58 vs 3.04+/-0.62, 2.23+/-0.58, P<0.01), the HSCs apoptotic index of HSCs (7.81+/-0.47 vs 1.63+/-0.25, 1.78+/-0.4, P<0.05) and the ratio of G0-G1 phase cells increased (94.30+/-1.33 vs 62.27+/-17.96, 50.53+/-2.25, P<0.05). The ratios of S-phase cells (3.11+/-1.27 vs 9.83+/-1.81, 11.87+/-1.9, P<0.05) and G2-M phase cells (2.58+/-0.73 vs 23.26+/-10.95, 13.60+/-1.15, P<0.01) declined., Conclusion: DSHX capsule shows certain therapeutic effects on hepatic fibrosis in rats and inhibits abnormal deposition of COL I and III in rat livers by promoting the apoptosis of HSCs and preventing their proliferation.

Background: Hepatic fibrosis is the common pathological change in various chronic liver diseases, and its major cause is the imbalance between the production and degradation of the extracellular matrix, which is mainly composed of collagens. Dan-Shao-Hua-Xian (DSHX) capsule, a traditional Chinese herbal compound, has shown marked preventive effects on hepatic fibrosis in rats in our previous studies. The present study was designed to further investigate its therapeutic actions on hepatic fibrosis in rats and its possible mechanisms., Methods: Eighty male Wistar rats were randomly divided into normal control group, hepatic fibrosis group, non-DSHX-treated group, low-dose-treated group, and high-dose-treated group. The rat models of hepatic fibrosis were established by subcutaneous injecton of CCl4, drinking alcohol, giving diet of hyperliprosis and hypoprotein for 8 weeks. The two DSHX-treated groups were treated respectively with low dose (0.5 g/kg) and high dose (1.0 g/kg) of DSHX capsule p.o. everyday for 8 weeks. At the end of the experiment, liver indexes were calculated in each group in addition to the levels of the serum hyaluronic acid and alanine aminotransferase. Their degree of hepatic fibrosis and urinary excretion of hydroxyproline and expression of collagen I, III were detected., Results: Comparison of the indexes of the hepatic fibrosis group and non-DSHX-treated group revealed that the liver indexes, levels of serum hyaluronic acid and alanine aminotransferase, and stage of hepatic fibrosis were all significantly reduced in the two DSHX treated groups. The urinary excretion of hydroxyproline was increased and the expression of collagen I and III in liver tissue was lessened. These alterations were more obviously observed in the high-dose-treated group., Conclusion: DSHX capsule has certain therapeutic effect on hepatic fibrosis in rats.