Qingda Granules alleviate brain damage in spontaneously hypertensive rats by modulating the miR-124/STAT3 signaling axis

doi:10.12122/j.issn.1673-4254.2025.01.03

Abstract

Abstract:

Objective To explore the mechanism of Qingda Granules (QDG) for alleviating brain damage in spontaneously hypertensive rats (SHRs). Methods Twelve 5-week-old SHRs were randomized into SHR control group and SHR+QDG group treated with QDG by gavage at the daily dose of 0.9 g/kg for 12 weeks. The control rats, along with 6 age-matched WKY rats, were treated with saline only. Blood pressure changes of the rats were monitored, and pathologies and neuronal apoptosis in the cerebral cortex were examined with HE staining and TUNEL staining. Cerebral cortical expressions of miR-124 and STAT3 mRNA were detected using RT-qPCR, and the protein expressions of NeuN, STAT3, Bcl-2, Bax, and cleaved caspase-3 were detected with immunohistochemistry and Western blotting. In a HT22 cell model of oxygen and glucose deprivation/reoxygenation (OGD/R), the effects of QDG on cell viability and apoptosis, expressions of miR-124 and STAT3 mRNA, and protein expressions of STAT3, Bcl-2, Bax, and cleaved caspase-3 were evaluated using CCK8 assay, Hoechst 33342 staining, RT-qPCR, and Western blotting. Results Compared with WKY rats, SHRs had significantly elevated systolic blood pressure, diastolic blood pressure and mean arterial pressure with significantly increased neuronal apoptosis in the cerebral cortex, reduced expressions of NeuN, miR-124 and Bcl-2, and enhanced expressions of STAT3, Bax and cleaved caspase-3 (P<0.05). All these changes in the SHRs were significantly ameliorated by treatment with QDG (P<0.05). In the HT22 cell model, QDG treatment obviously reduced OGD/R-induced cell apoptosis, increased the expressions of miR-124 and Bcl-2, and suppressed the elevation of protein expressions of STAT3, Bax and cleaved caspase-3. Conclusion QDG inhibits cerebral cortical neuronal apoptosis and thereby attenuates brain damage in SHR rats by modulating the miR-124/STAT3 signaling axis.

Key words: Qingda Granules, hypertension, brain damage, neurons, miR-124/STAT3 signaling axis

Qiaoyan CAI, Yaoyao XU, Yuxing LIN, Haowei LIN, Junpeng ZHENG, Weixiang ZHANG, Chunyu ZHAO, Yupeng LIN, Ling ZHANG. Qingda Granules alleviate brain damage in spontaneously hypertensive rats by modulating the miR-124/STAT3 signaling axis[J]. Journal of Southern Medical University, 2025, 45(1): 18-26.

Figures/Tables 7

Fig. 1 Effect of Qingda Granules (QDG) treatment on systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial pressure (MAP) in spontaneously hypertensive rats (SHRs). *P<0.05 vs WKY group; #P<0.05 vs SHR group.

Fig.2 Effects of Qingda Granules on pathological changes and neuron number in the cerebral cortex of SHRs. A: HE staining of the cortical area of the brain. B, C: Immunohistochemistry for detecting the protein expression of NeuN. Arrows indicate neuronal shrinkage. *P<0.05 vs WKY group; #P<0.05 vs SHR group.

Fig.3 Effects of Qingda Granules on neuronal apoptosis and protein expressions of Bcl-2, Bax and cleaved caspase-3 in the cerebral cortex of SHRs. A: TUNEL staining for detecting neuronal apoptosis. B: Immunohistochemistry for detecting protein expression of Bcl-2, Bax and cleaved caspase-3. *P<0.05 vs WKY group; #P<0.05 vs SHR group.

Fig. 4 Effect of Qingda Granules on expressions of miR-124 and STAT3 in the cerebral cortex of SHR rats. A: RT-qPCR analysis of mRNA expression of miR-124 and STAT3. B: Western blotting for detecting protein expressions of STAT3. *P<0.05 vs WKY group; #P<0.05 vs SHR group.

Fig.5 Effect of Qingda Granules on viability of HT22 cells with OGD/R. A: CCK-8 assay of viability of HT22 cells treated with different concentrations of Qingda Granules. B: CCK-8 assay of viability HT22 cells with OGD/R and Qingda Granules treatment. *P<0.05 vs 0 μg/mL group; #P<0.05 vs OGD/R group.

Fig.6 Effect of Qingda Granules on apoptosis and expressions of Bcl-2, Bax and cleaved caspase-3 in OGD/R-induced HT22 cells. A: Hoechst 33342 staining for detecting cell apoptosis. B: RT-qPCR analysis of mRNA expressions of Bcl-2 and Bax. C: Western blotting for detecting protein expressions of Bcl-2, Bax and cleaved caspase-3. *P<0.05 vs Control group; #P<0.05 vs OGD/R group.

Fig.7 Effect of Qingda Granules on expression of miR-124 and STAT3 in OGD/R-induced HT22 cells. A: RT-qPCR analysis of mRNA expression of miR-124 and STAT3. B: Western blotting for detecting STAT3 protein expression. *P<0.05 vs Control group; #P<0.05 vs OGD/R group.

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