摘要
目的 观察自噬在IL-1β促进INS-1胰岛β细胞凋亡中的作用。方法 体外培养大鼠INS-1胰岛β细胞株,给予不同浓度IL-1β干预,用CCK-8法检测在不同作用时间点INS-1细胞的存活率;用Western blot技术检测不同IL-1β浓度下自噬相关蛋白LC3、Beclin1、P62的表达水平;Western blot法和流式细胞术检测INS-1细胞的凋亡水平。结果 IL-1β可以降低INS-1细胞的存活率,且呈浓度及时间依赖性( P <0.05, P <0.01)。与对照组相比,随着IL-1β浓度的增高,INS-1细胞凋亡水平逐渐升高( P <0.05),且细胞自噬水平也随着IL-1β浓度的增高而逐渐增高(自噬相关蛋白LC3、Beclin1表达上升,P62降低, P <0.05)。采用自噬激动剂雷帕霉素预处理细胞上调自噬可进一步增加IL-1β诱导的细胞凋亡,而用自噬抑制剂3-MA预处理抑制细胞自噬后则可逆转IL-1β诱导的INS-1胰岛细胞凋亡。结论 IL-1β通过诱导自噬增加促进INS-1胰岛β细胞凋亡。
Aim To investigate the role of autophagy in IL-1β-induced apoptosis of islet β cells. Methods Rat β cell line INS-1 was cultured in vitro . The survival rate of INS-1 cells was measured by CCK-8 method at different concentrations and time. The level of apoptosis was analyzed by flow cytometry and Western blot, and the expressions of autophagy-related proteins were detected by Western blot. Results IL-1β could reduce the survival rate of INS-1 cells in a concentration-and time-dependent manner ( P <0.05, P <0.01). Compared with control group, with the increase of IL-1β concentration, the level of apoptosis of INS-1 cells increased gradually ( P <0.05);at the same time, the expression of autophagy increased gradually ( P <0.05). Pretreatment with autophagy agonist rapamycin (RAPA) could increase autophagy ( P <0.05) and apoptotic level ( P <0.05). However, the expression of autophagy and apoptosis decreased after pretreatment with autophagy inhibitor 3-MA ( P <0.05). Conclusion IL-1β can promote the apoptosis of INS-1 cells by inducing autophagy.
作者
宋峰
王瑾
王丽
焦向英
SONG Feng;WANG Jin;WANG Li;JIAO Xiang-ying(Dept of Physiology,Basic Medical College, Shanxi Medical University, Taiyuan 030001, China;Dept of Pathology, Basic Medical College, Shanxi Medical University, Taiyuan 030001, China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2019年第11期1564-1569,共6页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 31401006)
山西省青年科技研究基金资助项目(No 201801D221401)
