清达颗粒通过调控miR-124/STAT3信号轴减轻自发性高血压大鼠脑损伤

doi:10.12122/j.issn.1673-4254.2025.01.03

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (1): 18-26.doi: 10.12122/j.issn.1673-4254.2025.01.03

清达颗粒通过调控miR-124/STAT3信号轴减轻自发性高血压大鼠脑损伤

蔡巧燕¹^,²^,³^,⁴(), 许瑶瑶¹, 林雨星¹, 林浩伟¹, 郑俊鹏¹, 张伟祥¹, 赵春雨¹, 林育鹏¹, 张铃¹^,²^,³^,⁴()

^1.福建中医药大学，中西医结合学院，福建福州 350122
^2.福建中医药大学，中西医结合研究院，福建福州 350122
^3.福建中医药大学，福建省中西医结合老年性疾病重点实验室，福建福州 350122
^4.福建中医药大学，陈可冀学术思想传承工作室，福建福州 350122

收稿日期:2024-06-21 出版日期:2025-01-20 发布日期:2025-01-20
通讯作者: 张铃 E-mail:cqy2005899@163.com;remona1986@126.com
作者简介:蔡巧燕，博士，高级实验师，E-mail: cqy2005899@163.com
基金资助:
国家自然科学基金(82374282);福建省自然科学基金(2022J01876);福建中医药大学陈可冀中西医结合发展基金(CKJ2023003);省级大学生创新创业训练计划项目(S202210393016);福建中医药大学青年科技创新培育计划项目(XQC2024004)

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

Qiaoyan CAI¹^,²^,³^,⁴(), Yaoyao XU¹, Yuxing LIN¹, Haowei LIN¹, Junpeng ZHENG¹, Weixiang ZHANG¹, Chunyu ZHAO¹, Yupeng LIN¹, Ling ZHANG¹^,²^,³^,⁴()

^1.College of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China
^2.Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China
^3.Fujian Provincial Key Laboratory of Integrative Medicine on Geriatrics, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China
^4.Chen Keji Academic Thought Inheritance Studio, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China

Received:2024-06-21 Online:2025-01-20 Published:2025-01-20
Contact: Ling ZHANG E-mail:cqy2005899@163.com;remona1986@126.com
Supported by:
National Natural Science Foundation of China(82374282)

摘要/Abstract

摘要：

目的探讨清达颗粒（QDG）对自发性高血压大鼠（SHR）脑损伤的保护作用及其对miR-124/STAT3信号轴的影响。方法 6只5周龄WKY大鼠为WKY组，12只5周龄SHR大鼠分为SHR组、SHR+QDG组，6只/组。SHR+QDG组大鼠每天给予QDG灌胃，灌胃剂量为0.9 g/（kg·d），连续灌胃12周，其余组大鼠每天给予等体积生理盐水灌胃。利用鼠尾无创血压仪监测血压，HE染色观察脑皮质区病理，TUNEL染色检测皮质区凋亡，Q-PCR检测miR-124、STAT3 mRNA的表达，免疫组化和Western blotting检测NeuN、STAT3、Bcl-2、Bax、cleaved caspase-3的表达；利用氧糖剥夺/复氧（OGD/R）构建神经元HT22细胞损伤模型，分为Control组、QDG组、OGD/R组、OGD/R+QDG组。利用CCK-8法检测细胞活力，Hoechst 33342染色检测细胞凋亡，Q-PCR检测miR-124、STAT3、Bcl-2、Bax mRNA的表达，Western blotting检测STAT3、Bcl-2、Bax、cleaved caspase-3的表达。结果体内实验中，与WKY组比较，SHR组大鼠的收缩压、舒张压及平均动脉压均升高（P<0.05），脑皮质区神经元出现核固缩，数量减少，凋亡率升高（P<0.05），NeuN、miR-124、Bcl-2的表达降低，STAT3、Bax、cleaved caspase-3的表达升高（P<0.05）。与SHR组比较，清达颗粒能降低SHR大鼠的收缩压、舒张压及平均动脉压（P<0.05），改善SHR大鼠脑皮质区的病理损伤，降低皮质区的凋亡率（P<0.05），升高皮质区的NeuN、miR-124、Bcl-2的表达（P<0.05），降低STAT3、Bax、cleaved caspase-3的表达（P<0.05）。体外实验中，与Control组比较，OGD/R组凋亡率升高，miR-124、Bcl-2表达降低，STAT3、Bax、cleaved caspase-3表达升高（P<0.05）；与OGD/R组比较，清达颗粒能降低OGD/R诱导的HT22细胞凋亡，升高miR-124、Bcl-2表达，降低STAT3、Bax、cleaved caspase-3表达（P<0.05）。结论清达颗粒可能通过调控miR-124/STAT3信号轴，抑制神经元的凋亡，从而减轻SHR大鼠的脑损伤。

关键词: 清达颗粒, 高血压, 脑损伤, 神经元, miR-124/STAT3信号轴

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

蔡巧燕, 许瑶瑶, 林雨星, 林浩伟, 郑俊鹏, 张伟祥, 赵春雨, 林育鹏, 张铃. 清达颗粒通过调控miR-124/STAT3信号轴减轻自发性高血压大鼠脑损伤[J]. 南方医科大学学报, 2025, 45(1): 18-26.

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.

图/表 7

图1 清达颗粒对SHR大鼠收缩压、舒张压及平均动脉压的影响

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.

图2 清达颗粒对SHR大鼠脑皮质区病理改变及神经元数量的影响

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.

图3 清达颗粒对SHR大鼠脑皮质区凋亡及Bcl-2、Bax及cleavedcaspase-3蛋白表达的影响

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.

图4 清达颗粒对SHR大鼠脑皮质区miR-124、STAT3表达的影响

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.

图5 清达颗粒对OGD/R诱导的HT22细胞活力的影响

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.

图6 清达颗粒对OGD/R诱导的HT22细胞凋亡及Bcl-2、Bax及cleaved caspase-3表达的影响

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.

图7 清达颗粒对OGD/R诱导的HT22细胞中miR-124、STAT3表达的影响

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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清达颗粒通过调控miR-124/STAT3信号轴减轻自发性高血压大鼠脑损伤

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

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