二氢杨梅素通过下调JNK信号拮抗高糖诱导的PC12细胞凋亡  

Dihydromyricetin Inhibits High Glucose Induced PC12 Cells Apoptosis by Down-regulating JNK Pathway

作　　者：吕慧婕 朱责梅 陈维昭 何剑琴[1] 杨丝丝[1] 张恺芳[1] 奉水东[4] 凌宏艳[1] LU Hui-Jie;ZHU Ze-Mei;CHEN Wei-Zhao;HE Jian-Qin;YANG Si-Si;ZHANG Kai-Fang;FENG Shui-Dong;LING Hong-Yan(Department of Physiology, University of South Chino, Hengyang 421001, China;Department of Medicine, Changde Vocational Technical College, Changde 415003, China;Class 1 of 2015 Excellent Doctor, University of South Chino, Hengyang421001, China;Department of Social Medicine and Health Management, University of South China, Hengyang 421001, China)

机构地区：[1]南华大学生理学教研室,衡阳421001 [2]常德职业技术学院医学系,常德415003 [3]南华大学2015级卓越医生1班,衡阳421001 [4]南华大学社会医学与卫生管理教研室,衡阳421001

出　　处：《生物化学与生物物理进展》2018年第6期663-671,共9页Progress In Biochemistry and Biophysics

基　　金：湖南省自然科学基金(2018JJ2347); 湖南教育厅基金(16C1411); 衡阳市科技局基金(2016KJ64)资助

摘　　要：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(15μmol/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）. Hoechst33258 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 and96 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.

关 键 词：二氢杨梅素 高糖 PC12细胞凋亡 JNK信号 

分 类 号：R587[医药卫生—内分泌] R742[医药卫生—内科学]