南华大学生理学教研室,常德职业技术学院医学系,南华大学级卓越医生班,南华大学生理学教研室,南华大学生理学教研室,南华大学生理学教研室,南华大学社会医学与卫生管理教研室,南华大学生理学教研室
湖南省自然科学基金(2018JJ2347),湖南教育厅基金(16C1411)和衡阳市科技局基金(2016KJ64)资助
Department of Physiology,University of South China,Department of Medicine,Changde Vocational Technical College,Class of excellent doctor,University of South China,Department of Physiology,University of South China,Department of Physiology,University of South China,Department of Physiology,University of South China,Department of Social Medicine and Health Management, University of South China,Department of Physiology,University of South China
This work was supported by grants from Hunan Provincial Natural Science Foundation (2018JJ2347) , Hunan Education Department Foundation(16C1411) and Hengyang Municipal Bureau of Science and Technology Foundation(2016KJ64)
本文探讨二氢杨梅素(dihydromyricetin,DHM)是否通过下调JNK信号拮抗高糖诱导的PC12细胞凋亡.使用四甲基偶氮唑盐法(MTT)检测PC12细胞活力;流式细胞仪检测PC12细胞早、晚期凋亡及死亡细胞率;Hoechst 33258染色观察凋亡细胞核变化;蛋白质印迹法(Western blotting)检测PC12细胞凋亡相关蛋白(Bax、Bcl-2、cleaved-Caspase-3)和p-JNK蛋白的表达.结果发现,不同浓度的葡萄糖(4.5、9.0、13.5、18.0 g/L)分别处理PC12细胞24、48、72、96 h后,发现浓度为13.5 g/L的高糖处理PC12细胞72 h可明显改变细胞形态、降低细胞活力、增加细胞凋亡率,同时促凋亡蛋白(Bax、Caspase-3)表达增加、抗凋亡蛋白Bcl-2表达降低,提示:长时间高糖处理可诱导PC12细胞凋亡.DHM(15 ?滋mol/L)预处理能明显改善高糖诱导的PC12细胞凋亡,降低高糖诱导的PC12细胞中JNK和p-JNK蛋白的表达;进一步用JNK激动剂(茴香霉素)处理能取消DHM对高糖诱导PC12细胞凋亡的保护作用.综上,得出结论:DHM通过下调JNK信号拮抗高糖诱导的PC12细胞凋亡.
To investigate whether dihydromyricetin(DHM)inhibits high glucose induced PC12 cell apoptosis by downregulating JNK signaling. The cell viabilities of PC12 cells were assessed by MTT assay. The apoptotic rates of PC12 cells were measured after Annexin-V/PI (propidium iodide) staining by flow cytometry (FCM). Hoechst 33258 staining was used to detect the morphology of apoptotic PC12 cells. The expression of apoptosis-related proteins (Bax, Bcl-2, cleaved-Caspase-3) and the level of p-JNK in PC12 cells were detected by Western blotting assay. After PC12 cells were treated with different concentrations of glucose (4.5, 9, 13.5, 18 g/L) at 24, 48, 72 and 96 h respectively, the results showed that 13.5 g/L high glucose treatment for 72 h could significantly change the cell morphology, reduce cell viability, increase the apoptosis rate, at the same time, the expression of pro-apoptotic protein (Bax, Caspase-3) was increased and anti-apoptotic protein Bcl-2 was decreased, indicating that long-term high glucose treatment induced PC12 cell apoptosis. However, pretreatment with DHM (15 μmol/l) could significantly improve high glucose induced PC12 cell apoptosis and decrease the expression of JNK and p-JNK high glucose induced PC12 cell. Further treatment with JNK agonists (Anisomycin), which could eliminate the protective effect of DHM on apoptosis induced by high glucose in PC12 cells. In conclusion, DHM antagonizes high glucose-induced PC12 cell apoptosis by down-regulating JNK signaling.
吕慧婕,朱责梅,陈维昭,何剑琴,杨丝丝,张恺芳,奉水东,凌宏艳.二氢杨梅素通过下调JNK信号拮抗高糖诱导的PC12细胞凋亡[J].生物化学与生物物理进展,2018,45(6):663-671
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