Search

To investigate the regulatory effect of miR-1298 on hypoxic-ischemic nerve injury by cell culture in vitro. Firstly, the modeling effect of oxygen glucose deprivation/reoxygenation (OGD/R) of rat PC-12 cells was determined by cell activity detection and lactate dehydrogenase (LDH) cytotoxicity. At the same time, the expression difference of miR-1298 was detected by real-time fluorescence quantitative PCR (RT-qPCR). The miR-1298mimic, mimic NC, miR-1298 inhibitor and inhibitor NC were transfected into rat PC-12 cell line in vitro. The transfection efficiency of miR, mimic NC, inhibitor and inhibitor NC was detected. After OGD/R treatment, the cells were divided into control group, OGD/R group, mimicgroup, mimicNC group, inhibitor group and inhibitor NC group. The apoptosis of PC-12 cells in each group was detected by flow cytometry, and the expression of PC-12 apoptosis related protein B lymphoma-2 gene (BCL-2) and Bcl-2 associated X protein (BAX) were detected by Western blot. After OGD/R treatment, the survival rate of PC12 cells decreased significantly compared with the control group, and the leakage rate of LDH increased significantly (all P<0.05). The relative expression of miR-1298 in model cells was significantly lower than that in control group (P<0.05). 24 hours after transfection, the relative expression of miR-1298 in mimic group cells was significantly higher than that in mimicNC group ( P<0.05). The cells apoptosis rate in mimic group was lower than that in mimicNC group, while cells apoptosis rate in inhibitor group was higher than that in inhibitor NC group (all P<0.05). The expression of BCL-2 in mimic group was higher than that in mimicNC group, while the expression of BAX decreased. Compared with the inhibitorNC group, the expression of BCL-2 decreased, while the expression of BAX increased, and the differences were statistically significant (all P<0.05). miR-1298 alleviates the injury of OGD/R in PC-12 cells by inhibiting apoptosis. [ABSTRACT FROM AUTHOR]

BACKGROUND: Previous studies have found that the culture supernatant of the olfactory ensheathing cells is capable of promoting axonal regeneration and functional recovery after spinal cord injury, but there is a lack of research in the field of peripheral nerve. OBJECTIVE: To investigate whether olfactory ensheathing cell culture supernatant is beneficial for nerve repair after peripheral nerve injury. METHODS: Olfactory ensheathing cells were isolated and purified, to prepare the supernatant. The olfactory ensheathing cell culture supernatant was applied to the dorsal root ganglion tissue block in vitro to observe the axon growth of the dorsal root ganglion. The olfactory ensheathing cell culture supernatant was applied to a rat sciatic nerve defect model in vivo to examine its effect on axonal regeneration and myelinization of the injured nerve. RESULTS AND CONCLUSION: The purity of olfactory ensheathing cells was (94.4±3.1)%. Compared with the blank control and low dose olfactory ensheathing cell culture groups, the average length of five longest axons in dorsal root ganglion tissue mass in the high dose olfactory ensheathing cells culture group was significantly increased (P < 0.05). Immunofluorescence showed that the regenerated nerve penetrated through the defect area and the regenerated nerve was arranged orderly in the olfactory ensheathing cell culture and the autologous nerve groups, which was significantly superior to that in the blank control group. Transmission electron microscope observed that the number of regenerated nerve axons and the thickness of myelin sheath in the olfactory ensheathing cell culture group were significantly higher than those in the blank control group (P < 0.05). These results indicate that the supernatant of the olfactory ensheathing cells can promote axonal regeneration after peripheral nerve injury and the myelination of the regenerated axons, which provides a new olfactory ensheathing cells-based acellular therapy for peripheral nerve injury. [ABSTRACT FROM AUTHOR]