摘要
目的探讨抗纤益心方对扩张型心肌病大鼠Smad3,miR-23-3p,miR-145-5p表达的影响。方法将90只Wistar大鼠随机分为正常对照组(A组)、模型组(B组)、卡托普利组(C组)、抗纤益心方低剂量组(D组)、抗纤益心方中剂量组(E组)和抗纤益心方高剂量组(F组)。A组大鼠常规饲养,其余各组大鼠制备扩张型心肌病大鼠模型;制模成功后,B组每日灌胃卡托普利10.125 mg/kg,D组、E组、F组每日分别灌胃抗纤益心方4.7,9.4,18.8 mg/kg。药物干预8周后,检测大鼠心功能,取腹腔主动脉血,荧光定量聚合酶链式反应(PCR)法测定miR-23-3p和miR-145-5p的表达水平;取心肌组织,采用蛋白质印迹(WB)法测定Smad3蛋白表达;酶联免疫吸附试验(ELISA)法检测匀浆液转化生长因子β(TGF-β)、基质金属酶-1(MMP-1)及基质金属酶-1抑制剂(TIMP-1)蛋白水平。结果与A组相比,B组大鼠的左室收缩末压(LVESP)和左心室内压最大上升/下降速率(±dp/dtmax)明显下降,左室舒张末压(LVEDP)明显上升;与B组相比,C组、D组、E组、F组大鼠的LVESP和±dP/dtmax上升,LVEDP下降;与D组相比,E组和F组大鼠的LVESP和±d P/dtmax上升,LVEDP下降,差异均有统计学意义(P <0.05)。与A组相比,B组大鼠的心肌Smad3表达水平明显升高,循环miR-23-3p,miR-145-5p表达水平明显降低;与B组相比,C组、D组、E组、F组大鼠的心肌Smad3表达水平均明显降低,循环miR-23-3p,miR-145-5p表达水平均明显升高,差异均有统计学意义(P <0.05);与D组相比,E组和F组大鼠的循环miR-23-3p和miR-145-5p表达依次升高,Smad3表达依次降低,差异均有统计学意义(P <0.05),表明心肌Smad3表达水平与循环miR-23-3p,miR-145-5p表达水平呈负相关性(P <0.05)。与A组相比,B组大鼠的TGF-β和MMP-1表达水平明显升高,TIMP-1表达水平明显降低(P <0.01);与B组相比,C组、D组、E组、F组大鼠的TGF-β和MMP-1表达水平明显降低,TIMP-1表达水平明显升高;与D组相比,E组和F组大鼠的TGF-β和MMP-1水平均依次降低,TIMP-1水平依次升高,差异均有统计学意义(P <0.05)。结论抗纤益心方可上调扩张型心肌病大鼠miR-23-3p,miR-145-5p表达,下调Smad3表达,进而影响TGF-β/Smad3通路的功能,可能为其改善心室重构的作用机制之一。
Objective To investigate the effect of Kangxianyixin Prescription on the expression of Smad3,miR-23-3 p and miR-145-5 p in rats with dilated cardiomyopathy.Methods A total of 90 Wistar rats were randomly divided into normal control group(group A),model group(group B),captopril group(group C),low dose of Kangxianyixin Prescription group(group D),medium dose group of Kangxianyixin Prescription(group E)and high dose of Kangxianyixin Prescription group(group F).Group A was raised routinely,and the other groups were used to prepare dilated cardiomyopathy rat models.After the successful preparation of the model,group C was given a daily gavage of 10.125 mg/kg.Groups D,E,F were given a daily gavage of 4.7,9.4,and 18.8 mg/kg respectively.Eight weeks after the drug intervention,the cardiac function of the rats was detected,and abdominal aortic blood was taken.The expression levels of miR-23-3 p and miR-145-5 p were determined by fluorescence quantitative PCR.Myocardial tissue was taken and the protein expression of Smad3 was determined by Western blot.The enzyme-linked immunosorbent assay(ELISA)method was used to detect the protein levels of transforming growth factorβ(TGF-β),matrix metalloenzyme-1(MMP-1)and matrix metalloenzyme inhibitor(TIMP-1)in the homogenate.Results Compared with group A,the left ventricular end systolic pressure(LVESP)and maximum increase/decrease rate of left ventricular pressure(±dP/dtmax)in group B descended evidently and their left ventricular end diastolic pressure(LVEDP)increased obviously.Compared with group B,the LVESP and±d P/dtmaxin groups C,D,E,D,F increased obviously,and their LVEDP descended evidently.Compared with group D,the LVESP and±dP/dtmaxin groups E,F increased,and their LVEDP descended,and the difference were statistically significant(P<0.05).Compared with group A,the expression level of Smad3 in group B increased obviously,and their expression level of circulating miR-23-3 p and miR-145-5 p descended evidently.Compared with group B,the expression level of Smad3 in groups C,D,E,D,F descended evidently,and their expression level of circulating miR-23-3 p and miR-145-5 p increased obviously.Compared with group D,the miR-23-3 p and miR-145-5 p in groups E,F increased successively,and their Smad3 descended successively,and the difference were statistically significant(P<0.05).Upon examination,the expression level of myocardial Smads was negatively correlated with the expression level of circulating miR-23-3 p and miR-145-5 p(P<0.05).The results of ELISA showed that compared with group A,the expression level of TGF-βand MMP-1 in group B increased obviously,and its expression of TIMP-1 decreased(P<0.05).Compared with the group B,the expression level of TGF-βand MMP-1 in groups C,D,E,D,F descended evidently,and their expression level of TIMP-1 increased obviously.Compared with group D,the TGF-βand MMP-1 in groups E,F descended successively,and their TIMP-1 increased successively,and the difference were statistically significant(P<0.05).Conclusion Kangxianyixin Prescription could up-regulate the expression of circulating miR-23-3 p and miR-145-5 p,while down-regulate the expression of Smad3,thus influence TGF-β/Smad3 pathway in rats with dilated cardiomyopathy,which may be one of the mechanisms to improve ventricular remodeling.
作者
杨曦
熊剑
魏云杰
YANG Xi;XIONG Jian;WEI Yunjie(Department of Cardiovascular Medicine,Taihe Hospital·Affiliated Hospital of Hubei University of Medicine,Shiyan,Hubei,China 442000)
出处
《中国药业》
CAS
2020年第17期29-34,共6页
China Pharmaceuticals
作者简介
第一作者:杨曦,男,硕士研究生,住院医师,主要从事心血管方面的基础及临床研究工作,(电子信箱)850611782@qq.com;通信作者:魏云杰,男,硕士研究生,主治医师,主要从事心血管方面的基础及临床研究工作,(电子信箱)weiyunjie200@163.com。
