吴门调脂颗粒对大鼠平滑肌细胞自噬的干预研究  

Intervention of Wumen Tiaozhi Granules(吴门调脂颗粒)on Autophagy of Rat Smooth Muscle Cells

作　　者：吴瑛滢 张国兴[2] 陈竞纬[1] WU Ying-ying;ZHANG Guo-xing;CHEN Jing-wei(Suzhou Affiliated Hospital of Nanjing University of Chinese Medicine,Suzhou Jiangsu 215000,China;Soochow University,Suzhou Jiangsu 215000,China)

机构地区：[1]南京中医药大学苏州附属医院,江苏苏州215000 [2]苏州大学,江苏苏州215000

出　　处：《中医药导报》2020年第12期19-23,共5页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：苏州市2015年度第九批产业技术创新专项(应用基础研究·医疗卫生)(SYS201573)。

摘　　要：目的:探究吴门调脂颗粒对大鼠平滑肌细胞(VSMCs)的自噬在细胞、蛋白质、基因等水平的影响。方法:运用血清药理学方法,以调脂颗粒含药血清、6-氨基-3-甲基嘌呤(3-MA)、雷帕霉素(Rapamycin)诱导处理VSMCs,通过MTT检测调脂颗粒含药血清对细胞生长增殖的影响,Western blotting检测自噬相关蛋白LC3-Ⅱ的相对表达量变化,免疫荧光法检测细胞内自噬小体的数目变化,Real time PCR检测自噬相关基因表达变化。结果:调脂颗粒含药血清处理VSMCs后,细胞生存率无明显改变,自噬相关蛋白LC3-Ⅱ的相对表达上调,自噬小体数量增多,自噬相关基因表达上调(P<0.05)。雷帕霉素联合调脂颗粒含药血清处理VSMCs后,细胞生存率下降,较雷帕霉素组LC3-Ⅱ的相对表达下降,自噬小体数量显著减少,自噬相关基因表达下调(P<0.05或P<0.01)。结论:吴门调脂颗粒可以通过多个层面抑制平滑肌细胞的异常增殖及过度自噬,促进平滑肌细胞适度自噬。Objective: To explore the effects of Wumen Tiaozhi granules on the autophagy of rat smooth muscle cells(VSMCs) at cell, protein and gene levels. Methods: Using serum pharmacology methods, VSMCs were induced by Wumen Tiaozhi granules containing serum, 6-amino-3-methylpurine(3-MA) and rapamycin(Rapamycin). MTT was used to detect the effect of Tiaozhi granule-containing serum on cell growth and proliferation. Western blotting was used to detect the relative expression of autophagy-related protein LC3-Ⅱ. Immunofluorescence was used to detect the number of autophagosomes in the cell. Real time PCR was used to detect the expression of autophagy-related genes. Results: There was no significant change in the cell survival rate when the VSMCs were treated with Wumen Tiaozhi granules. At the same time, the relative expression of autophagy-related protein LC3-Ⅱ was up-regulated, the number of autophagosomes was significantly increased,and the expression of autophagy-related genes was up-regulated(P <0.05). The cell survival rate of VSMCs treated with Wumen Tiaozhi granules combined with rapamycin was decreased. Compared with the rapamycin group, the relative expression of LC3-Ⅱ was decreased, the number of autophagosomes was significantly reduced, and the expression of autophagy-related genes was down-regulated(P<0.05 or P<0.01). Conclusion: Wumen Tiaozhi granules can inhibit the abnormal proliferation and excessive autophagy of smooth muscle cells through multiple levels, and promote proper autophagy of smooth muscle cells.

关 键 词：自噬 吴门调脂颗粒 中医药 平滑肌细胞 心血管疾病 大鼠 

分 类 号：R285.5[医药卫生—中药学]