三参通脉合剂通过上调microRNA-146a改善大鼠心肌细胞H9C2的氧化损伤  

Sanshentongmai Mixture Improves Oxidative Damage in Rat Cardiomyocytes H9C2 via Upregulation of microRNA-146a

作　　者：李然[1] 王振裕[1] 王燕丽[1] 滕菲[1] LI Ran;WANG Zhenyu;WANG Yanli;TENG Fei(Department of Health Care for Cadres,Beijing Hospital of Traditional Chinese Medicine Affiliated to the Capital Medical University,Beijing 100010,China)

机构地区：[1]首都医科大学附属北京中医医院干部保健科,北京100010

出　　处：《四川大学学报（医学版）》2024年第3期630-634,共5页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(No.8150140342)资助。

摘　　要：目的 探讨三参通脉(Sanshentongmai, SSTM)合剂调节微小RNA-146a(microRNA-146a)对大鼠心肌细胞H9C2氧化损伤的影响及作用机制。方法 体外培养大鼠心肌细胞H9C2,过氧化氢(H_(2)O_(2))作为氧化剂制作H9C2氧化应激模型。通过三参通脉干预,观察H_(2)O_(2)诱导的H9C2细胞氧化损伤的变化以及microRNA-146a的表达,探讨三参通脉对H9C2的保护作用及其作用机制。将体外培养的H9C2分为空白组、H_(2)O_(2)氧化损伤模型组(简称模型组)、H_(2)O_(2)模型+三参通脉药物(500μg/mL,处理72 h)组(简称模型加药组)。通过细胞计数试剂盒CCK8检测细胞活力,酶联免疫吸附试验(enzyme-linked immunosorbent assay, ELISA)检测血清N端脑钠肽前体(N-terminal pro-brain natriuretic peptide, NtproBNP)、一氧化氮(nitric oxide, NO)、超敏C反应蛋白(high-sensitivity C-reactive protein, Hs-CRP)和血管紧张素Ⅱ(angiotensinⅡ)水平,实时荧光定量PCR(RT-PCR)检测系统检测microRNA-146a表达水平。结果 通过CCK8法检测细胞活力,发现在药物质量浓度为500μg/mL时,细胞增殖改善达到顶峰,故选用此浓度为干预浓度。ELISA检测的心衰相关指标:Nt-proBNP、NO、Hs-CRP、angiotensinⅡ水平中,与空白组比较,模型加药物组中Nt-proBNP、angiotensinⅡ表达上调(P<0.05),NO表达下调(P<0.05),Hs-CRP与空白组比较表达差异无统计学意义,说明三参通脉可以有效改善大鼠心肌细胞H9C2氧化损伤。最后,RT-PCR法检测microRNA-146a在各组的表达可以看出,15μmol/L H_(2)O_(2)处理可以明显降低microRNA-146a表达,而模型加药组中microRNA-146a表达较模型组升高(P<0.05),与空白组对比差异无统计学意义。结论 三参通脉可以明显抵抗H_(2)O_(2)诱导的H9C2细胞氧化损伤,可能是通过上调microRNA-146a从而发挥心肌保护作用。Objective To investigate the effect of Sanshentongmai(SSTM)mixture on the regulation of oxidative damage to rat cardiomyocytes(H9C2)through microRNA-146a and its mechanism.Methods H9C2 were cultured in vitro,H_(2)O_(2) was used as an oxidant to create an oxidative damage model in H9C2 cells.SSTM intervention was administered to the H9C2 cells.Then,the changes in H_(2)O_(2)-induced oxidative damage in H9C2 cells and the expression of microRNA-146a were observed to explore the protective effect of SSTM on H9C2 and its mechanism.H9C2 cells cultured in vitro were divided into 3 groups,including a control group,a model group of H_(2)O_(2)-induced oxidative damage(referred to hereafter as the model group),and a group given H_(2)O_(2) modeling plus SSTM intervention at 500μg/L for 72 h(referred to hereafter as the treatment group).The cell viability was measured by CCK8 assay.In addition,the levels of N-terminal pro-brain natriuretic peptide(Nt-proBNP),nitric oxide(NO),high-sensitivity C-reactive protein(Hs-CRP),and angiotensin were determined by enzyme-linked immunosorbent assay(ELISA).The expression level of microRNA-146a was determined by real-time PCR(RT-PCR).Result H9C2 cells were pretreated with SSTM at mass concentrations ranging from 200 to 1500μg/L.Then,CCK8 assay was performed to measure cell viability and the findings showed that the improvement in cell proliferation reached its peak when the mass concentration of SSTM was 500μg/L,which was subsequently used as the intervention concentration.ELISA was performed to measure the indicators related to heart failure,including Nt-proBNP,NO,Hs-CRP,and angiotensinⅡ.Compared with those of the control group,the expressions of Nt-proBNP and angiotensinⅡin the treatment group were up-regulated(P<0.05),while the expression of NO was down-regulated(P<0.05).There was no significant difference in the expression of Hs-CRP between the treatment group and the control group.These findings indicate that SSTM could effectively ameliorate oxidative damage in H9C2 rat cardiom

关 键 词：大鼠心肌细胞H9C2 三参通脉合剂 氧化损伤 微小RNA-146a 

分 类 号：R285.5[医药卫生—中药学]