丹参通络解毒汤调控METTL3介导的m^(6)A修饰对缺氧/复氧CMECs的保护作用  

Protective effects of Danshen Tongluo Jiedu Decoction on hypoxia/reoxygenation-injured CMECs through regulating METTL3-mediated m^(6)A modification

作　　者：陆哲雯 刘莉 蒋啸 李鑫辉 LU Zhewen;LIU Li;JIANG Xiao;LI Xinhui(School of Chinese Medicine,Hunan University of Chinese Medicine,Changsha,Hunan 410208,China)

机构地区：[1]湖南中医药大学中医学院,湖南长沙410208

出　　处：《湖南中医药大学学报》2025年第3期438-444,共7页Journal of Hunan University of Chinese Medicine

基　　金：国家自然科学基金面上项目(82074392);湖南省自然科学基金区域联合基金项目(2023JJ50036);湖南省中医药管理局重点项目(A2023013);湖南省教育厅科学研究项目(23A0283)。

摘　　要：目的探讨丹参通络解毒汤对缺氧/复氧(H/R)损伤大鼠心肌微血管内皮细胞(CMECs)的保护作用及甲基转移酶样3(METTL3)的影响。方法建立H/R损伤CMECs模型,将细胞分为空白组、模型组、中药组、过表达组、中药+过表达组、空载组,ELISA检测各组炎症因子IL-1β和IL-10水平,流式细胞术检测各组细胞凋亡情况,Western blot检测各组METTL3蛋白表达水平,比色法检测各组m^(6)A及肌酸激酶CK水平。结果与空白组比较,模型组METTL3蛋白表达与m^(6)A水平均升高(P<0.01),CMECs凋亡率升高(P<0.01),IL-1β、CK含量升高(P<0.01),IL-10含量降低(P<0.01);与模型组比较,中药组METTL3蛋白表达与m^(6)A水平均降低(P<0.05,P<0.01),CMECs凋亡率显著降低(P<0.01),IL-1β、CK含量显著降低(P<0.01),IL-10含量显著升高(P<0.01);与模型组比较,过表达组METTL3蛋白表达升高(P<0.01),CMECs凋亡率升高(P<0.01),IL-1β、CK含量升高(P<0.01,P<0.05),IL-10含量降低(P<0.01);与过表达组相比,中药+过表达组METTL3蛋白表达降低(P<0.01),CMECs凋亡率降低(P<0.01),IL-1β、CK含量降低(P<0.01,P<0.05),IL-10含量显著升高(P<0.01)。结论丹参通络解毒汤能降低炎症反应,抑制细胞凋亡,对缺氧/复氧CMECs有保护作用,其机制可能与抑制METTL3表达,从而下调m^(6)A水平有关。Objective To explore the protective effects of Danshen Tongluo Jiedu Decoction(DSTLJDD)on cardiac microvascular endothelial cells(CMECs)of rats with hypoxia/reoxygenation(H/R)injury and its influence on methyltransferase-like 3(METTL3).Methods The H/R-injured CMECs model was established,and the cells were divided into blank group,model group,Chinese medicine(CM)group,overexpression group,CM+overexpression group,and empty vector group.The levels of inflammatory factors IL-1β and IL-10 in each group were determined by ELISA,cell apoptosis was measured by flow cytometry,the protein expression level of METTL3 was checked by Western blot,and the content of m^(6)A and creatine kinase(CK)were examined by colorimetry.Results Compared with the blank group,the protein expression of METTL3 and the level of m^(6)A in the model group increased(P<0.01),the apoptosis rate of CMECs increased(P<0.01),the content of IL-1βand CK increased(P<0.01),while the content of IL-10 decreased(P<0.01).Compared with the model group,the protein expression of METTL3 and the level of m^(6)A in the CM group decreased(P<0.05,P<0.01),the apoptosis rate of CMECs markedly decreased(P<0.01),the content of IL-1β and CK significantly decreased(P<0.01),while the content of IL-10 notably increased(P<0.01).Compared with the model group,the protein expression of METTL3 in the overexpression group increased(P<0.01),the apoptosis rate of CMECs increased(P<0.01),the content of IL-1βand CK increased(P<0.01,P<0.05),while the content of IL-10 decreased(P<0.01).Compared with the overexpression group,the protein expression of METTL3 in the CM+overexpression group decreased(P<0.01),the apoptosis rate of CMECs decreased(P<0.01),the content of IL-1β and CK decreased(P<0.01,P<0.05),while the content of IL-10 significantly increased(P<0.01).Conclusion DSTLJDD can reduce inflammatory responses,inhibit cell apoptosis,and exert protective effects on H/R-injured CMECs.Its mechanism may be related to the suppression of METTL3 expression,thereby downregulating m^(6)A leve

关 键 词：丹参通络解毒汤 甲基转移酶样3 m^(6)A修饰 心肌缺血再灌注损伤 心肌微血管内皮细胞 

分 类 号：R285.5[医药卫生—中药学]