南蛇藤通过调控内质网应激途径抑制巨噬细胞损伤  

Inhibitory Effect of Celastrus orbiculatus on Macrophages Injury by Regulating Endoplasmic Reticulum Stress Pathway

作　　者：宋园园[1] 司艳红[1] 张敬军[1] 张颖[1] SONG Yuan-yuan;SI Yan-hong;ZHANG Jing-jun;ZHANG Ying(Taishan Medical University,Taian 271000,China)

机构地区：[1]泰山医学院,山东泰安271000

出　　处：《中药材》2019年第6期1387-1392,共6页Journal of Chinese Medicinal Materials

基　　金：山东省重点研发计划(2015GSF119008);山东省高校科研计划(J12LM01);泰安市科技发展计划(201440774-03);国家级大学生创新创业计划(201610439289)。

摘　　要：目的:验证南蛇藤通过调控内质网应激(ERS)途径减轻氧化低密度脂蛋白(Ox-LDL)诱导巨噬细胞损伤的作用机制。方法:体外培养RAW264.7巨噬细胞,给予50、100、150 mg/L南蛇藤提取物预处理30 min后加入100 mg/mL Ox-LDL损伤24 h,分别采用油红O染色和ELISA法检测细胞内脂质沉积情况和胆固醇含量,ELISA法和TBA法检测细胞内乳酸脱氢酶(LDH)活性和丙二醛(MDA)含量,流式细胞术检测细胞内活性氧簇(ROS)的释放,MTT法和Annexin V-FITC双染法检测细胞活力和凋亡情况,Western blot检测内质网应激相关蛋白CD36、GRP78、GRP94的表达。结果:南蛇藤浓度依赖性地减轻了Ox-LDL诱导的巨噬细胞内脂质蓄积和巨噬细胞的泡沫化,显著降低了Ox-LDL诱导的细胞LDH及ROS释放,下调了MDA水平,并抑制了Ox-LDL诱导的清道夫受体CD36与葡萄糖调节蛋白78/94(GRP78/94)的高表达。结论:南蛇藤可保护Ox-LDL导致的细胞损伤,其机制可能与调控内质网应激途径有关。Objective:To investigate the mechanism of Celastrus orbiculatus in reducing oxidative low-density lipoprotein(Ox-LDL)-induced macrophages injury by regulating endoplasmic reticulum stress(ERS).Methods:RAW264.7 cells were cultured in vitro and pretreated with Celastrus orbiculatus at different concentrations(50,100,150 mg/L)for 30 min and then treated with Ox-LDL(100 mg/mL)for 24 h.Lipids diposition and cholesterol content in the RAW264.7 macrophages were detected by oil red O staining and ELISA methods,the activity of lactate dehydrogenase(LDH)and the content of malondialdehyde(MDA)were measured by ELISA and TBA methods.The release of reactive oxygen species(ROS)was detected by flow cytometry,the cell viability and the cell apoptosis were examined with MTT and Annexin V-FITC double-staining methods,the expressions of CD36,GRP78,GRP94 were determined by Western blot.Results:Celastrus orbiculatus attenuated Ox-LDL-induced cholesterol accumulation in macrophages and foam cell formation in a content-dependent manner and significantly reduced the Ox-LDL-induced LDH and ROS releases and down-regulated MDA level in the macrophages,inhibited the high expression of scavenger receptor CD36 and GRP78,GRP94 induced by Ox-LDL.Conclusion:Celastrus orbiculatus can protect the cell damage caused by Ox-LDL,and its mechanism may be related to the regulation of endoplasmic reticulum stress pathway.

关 键 词：南蛇藤 内质网应激 氧化低密度脂蛋白 巨噬细胞损伤 

分 类 号：R285.5[医药卫生—中药学]