Neuroprotective effects of miR-30c on rats with cerebral ischemia/reperfusion injury by targeting SOX9.

2020 
OBJECTIVE To explore the neuroprotective effect and mechanism of miR-30c in rats with cerebral ischemia/reperfusion (CI/R) injury. METHODS in vivo, miR-30c mimic was transfected before the rats were treated with CI/R injury, and the neurological damage was evaluated. The expression of miR-30c was analyzed by RT-PCR.The area of cerebral infarction and pathological damage in CI/R rats were detected by TTC, H&E, Nissl and TUNEL staining. The expression of SOX9, BNDF and NT-3 were analyzed by immunofluorescence and western blot. The target relationship between SOX9 and miR-30c was analyzed by dual- luciferase reporter assay. in vitro, miR-30c mimic, SOX9 mimic and negative control were transfected before the OGD/R model of HT22 was established. CCK8 and flow cytometry were used to analyze the effect of miR-30c on the cell viability and apoptosis of HT22 cells in OGD/R condition. The protein expression of SOX9, BNDF and NT-3 were analyzed by immunofluorescence and western blot. RESULTS The expression of miR-30c in the CI/R rats were significantly lower than that of sham group (p < 0.05). The area of cerebral infarction and the number of pathological damage and apoptosis in hippocampus were improved by miR-30c. miR-30c reduced SOX9 expression and improved BNDF and NT-3 expression in hippocampus compared with CI/R group (p < 0.05). The results of dual-luciferase reporter showed that SOX9 was the target of miR-30c. Under OGD/R condition, miR-30c mimic promoted cell viability and reduced cell apoptosis. miR-30c decreased the expression of SOX9, which was consistent with the results of immunofluorescence. Moreover, miR-30c could increase the expression of BNDF and NT-3 in HT22 cells. After transfection of SOX9 mimic, it was found that the above effects of miR-30c were all reversed. CONCLUSION miR-30c can improve the pathological damage of rats with CI/R injury and plays a neuroprotective role both in vivo and in vitro by targeting SOX9.
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