健脾祛痰和血方通过调控线粒体自噬对氧化低密度脂蛋白诱导的巨噬细胞损伤保护作用机制  

Mechanism of Jianpi Qutan Hexue Decoction(健脾祛痰和血方)on Oxidative Low-Density Lipoprotein Induced Macrophage Injury by Regulating Mitophagy

作　　者：杨茗茜[1] 姜钧文[2] 袁东超[1] 李思琦[2] 季康寿 YANG Mingqian;JIANG Junwen;YUAN Dongchao;LI Siqi;JI Kangshou(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁中医药大学附属医院,辽宁沈阳110032

出　　处：《中华中医药学刊》2023年第11期74-77,I0016,I0017,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81874372);辽宁省自然科学基金项目(2019-ZD-0963,2023-MS-227);辽宁中医药大学基金项目(2021LZY025)。

摘　　要：目的探讨健脾祛痰和血方通过线粒体自噬对氧化低密度脂蛋白(ox-LDL)诱导的巨噬细胞损伤保护作用的机制研究。方法制备健脾祛痰和血方含药血清并进行分组:对照组、ox-LDL组、健脾祛痰和血方(JPQTHXD)低剂量组、JPQTHXD中剂量组与JPQTHXD高剂量组。CCK-8法检测不同处理对RAW 264.7细胞的增殖影响;细胞免疫荧光检测LC3的表达;ROS检测不同组别细胞内活性氧的含量;RT-RCR及Western blot检测PINK1、Parkin、LC3基因及蛋白的表达。结果健脾祛痰和血方可显著减轻ox-LDL诱导后的RAW 264.7细胞损伤,可降低细胞内LC3与ROS含量;健脾祛痰和血方可以显著逆转ox-LDL引起的PINK1、Parkin与LC3 mRNA与蛋白表达的变化。结论健脾祛痰和血方对ox-LDL诱导的巨噬细胞损伤具有一定的保护作用,其机制主要是通过线粒体自噬介导。Objective To investigate the mechanism of the protective effect of Jianpi Qutan Hexue Decoction(健脾祛痰和血方,JPQTHXD)on oxidized low density lipoprotein(ox-LDL)induced macrophage injury through mitophagy.Methods The serum containing JPQTHXD was prepared and divided into control group,ox-LDL group,JPQTHXD low-dose group,JPQTHXD medium-dose group and JPQTHXD high-dose group.The effects of different treatments on RAW 264.7 cell proliferation were detected by CCK-8 method.The expression of LC3 was detected by cell immunofluorescence.ROS was used to detect the content of reactive oxygen species(ROS)in different groups.RT-RCR and Western blot were used to detect the expressions of PINK1,Parkin,LC3 genes and proteins.Results JPQTHXD can significantly reduce the damage of RAW 264.7 cells induced by ox-LDL,and reduce the intracellular LC3 and ROS contents.JPQTHXD can significantly reverse the mRNA and protein expression changes of PINK1,Parkin and LC3 induced by ox-LDL.Conclusion JPQTHXD can protect ox-LDL induced macrophage injury,and the mechanism is mainly mediated by mitophagy.

关 键 词：健脾祛痰和血方 线粒体自噬 巨噬细胞 

分 类 号：R285.5[医药卫生—中药学]