Neuroprotective effects of miR-30c on rats with cerebral ischemia/reperfusion injury by targeting SOX9
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Min Zhang1, Yingjun Zhu1, Meng Wei1
1Department of Anesthesiology, The Affiliated Huaian No.1 People's Hospital of Nanjing Medical University, Huaian, 223300, Jiangsu, China.
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.
Insights
MicroRNA-30c (miR-30c) demonstrates significant neuroprotective effects against cerebral ischemia/reperfusion (CI/R) injury in rats. By targeting SOX9, miR-30c reduces brain damage and improves outcomes, offering a potential therapeutic strategy for stroke.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Cerebral ischemia/reperfusion (CI/R) injury is a major cause of stroke-related disability.
- MicroRNAs (miRNAs) play crucial roles in regulating cellular processes following ischemic events.
- The specific role of miR-30c in CI/R injury requires further elucidation.
Purpose of the Study:
- To investigate the neuroprotective effect of miR-30c in a rat model of CI/R injury.
- To elucidate the underlying molecular mechanism of miR-30c's action, focusing on its interaction with SOX9.
- To assess the impact of miR-30c on neuronal survival, apoptosis, and the expression of key neurotrophic factors.
Main Methods:
- In vivo studies involved transfecting miR-30c mimic in CI/R-induced rats and evaluating neurological damage, infarct size, and apoptosis.
- In vitro experiments utilized an oxygen-glucose deprivation/reperfusion (OGD/R) model in HT22 cells to assess cell viability and apoptosis.
- Gene expression analysis (RT-PCR, Western blot, immunofluorescence) and dual-luciferase reporter assays were employed to determine miR-30c and SOX9 levels and their interaction.
Main Results:
- miR-30c expression was significantly downregulated in CI/R rats.
- Overexpression of miR-30c reduced infarct volume, pathological damage, and apoptosis in the hippocampus of CI/R rats.
- miR-30c directly targeted SOX9, decreasing its expression while upregulating BDNF and NT-3.
- In vitro, miR-30c enhanced HT22 cell viability and reduced apoptosis under OGD/R conditions, effects reversed by SOX9 mimic transfection.
Conclusions:
- miR-30c exerts significant neuroprotective effects against CI/R injury in both in vivo and in vitro models.
- The mechanism involves the direct targeting of SOX9, leading to reduced neuronal damage and apoptosis.
- miR-30c represents a promising therapeutic target for mitigating the consequences of cerebral ischemia/reperfusion injury.
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