补阳壮通饮通过调控细胞增殖和线粒体自噬保护星形胶质细胞机制研究  

Study on the protective mechanism of Buyang Zhuangtong Yin on astrocytes by regulating cell proliferation and mitochondrial autophagy

作　　者：陈真珍[1] 李日著[2] 谢福周 林富生 李岩 王凯华[4] 黄龙坚 CHEN Zhenzhen;LI Rizhu;XIE Fuzhou;LIN Fusheng;LI Yan;WANG Kaihua;HUANG Longjian(Guangxi University of Chinese Medicine,Nanning 530200,China)

机构地区：[1]广西中医药大学,广西南宁530200 [2]右江民族医学院附属医院,广西百色533000 [3]右江民族医学院,广西百色533000 [4]广西中医药大学附属国际壮医医院,广西南宁530201 [5]广西高校右江流域特色民族药研究重点实验室,广西百色533000

出　　处：《陕西中医》2024年第12期1604-1608,共5页Shaanxi Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(82360870);广西自然科学基金资助项目(2021GXNSFAA196012,2023GXNSFAA026404);广西重点研发计划项目(桂科AB24010222);广西壮族自治区中医药管理局自筹科研项目(GXZYZ20210103);广西中医药大学科研项目(2021MS011)。

摘　　要：目的:初步探讨补阳壮通饮对脑缺血再灌注损伤(CIRI)的神经保护作用机制。方法:建立糖氧剥夺/再灌注(OGD/Rep)细胞模型,分为正常组(NC)、低剂量模型对照组(OGD-L)、中剂量模型对照组(OGD-M)、高剂量模型对照组(OGD-H)、补阳壮通饮低剂量组(BYZTY-L)、补阳壮通饮中剂量组(BYZTY-M)和补阳壮通饮高剂量组(BYZTY-H)。大鼠连续灌胃补阳壮通饮7 d以制备含药血清。利用含药血清对上述分组细胞进行干预培养3 d。MTT法检测细胞增殖活力;流式细胞仪检测细胞凋亡率变化;ELISA法检测炎症指标(TNF-α、IL-6)变化;Western blot检测线粒体自噬相关蛋白LC3、Beclin-1、P62和磷酸化p38-MAPK蛋白表达变化。结果:补阳壮通饮干预可增强模型组大鼠原代皮层神经细胞的增殖活力,抑制其凋亡,促进LC3、Beclin-1和磷酸化p38-MAPK蛋白的表达,抑制P62蛋白表达,且表现一定的剂量和时间效应。结论:补阳壮通饮对OGD/Rep损伤星形胶质细胞具有保护作用,机制可能与增强细胞增殖活力和线粒体自噬、抑制凋亡和炎症反应有关,p38-MAPK通路也可能参与其中。Objective:This study aims to explore the molecular mechanism of the neuroprotective effect of Buyang Zhuangtong Yin on cerebral ischemia-reperfusion injury.Methods:Established oxygen and glucose deprivation/reperfusion(OGD/Rep)cell models,and they were divided into normal group(NC),low-dose model control group(OGD-L),medium dose model control group(OGD-M),high-dose model control group(OGD-H),low-dose Buyang Zhuangtong Yin group(BYZTY-L),medium dose Buyang Zhuangtong Yin group(BYZTY-M),and high-dose Buyang Zhuangtong Yin group(BYZTY-H).Rats were continuously gavaged with Buyang Zhuangtong Yin for 7 days to prepare medicated serum.Using medicated serum to intervene and cultivate the above-mentioned grouped cells for 3 days.MTT assay was used to detect cell proliferation activity.Flow cytometry was used to detect changes in cell apoptosis rate.ELISA was used to detect the inflammatory markers(TNF-αand IL-6)changes.Western blot was used to detect the mitochondrial autophagy related proteins(LC3,Beclin-1,P62 and p-p38-MAPK)changes.Results:Buyang Zhuangtong Yin can enhance the proliferation activity of primary cortical neurons,inhibit apoptosis,and induce the expression of LC3,Beclin-1 and p38-MAPK,and inhibit the expression of P62,exhibit a certain dose and time effects.Conclusion:Buyang Zhuangtong Yin has a protective effect on astrocytes damaged by OGD/Rep injury,which may be related to enhancing cell proliferation activity and mitochondrial autophagy,inhibiting apoptosis and inflammatory response,and also the p38-MAPK pathway may also be involved.

关 键 词：补阳壮通饮 糖氧剥夺/再灌注 含药血清 抗炎 线粒体自噬 P38-MAPK 

分 类 号：R285.5[医药卫生—中药学]