红芪多糖联合厄贝沙坦对2型糖尿病大鼠心肌纤维化的影响

The Effects of Hedysarum Polybotys Saccharide Joined with Irbesartan on Myocardial Fibrosis of Type Two Diabetic Rats

目的:探讨红芪多糖单用或联合厄贝沙坦对2型糖尿病大鼠心肌纤维化的影响。方法:采用高脂高糖饲料加单次腹腔注射链脲佐菌素[30 mg/kg(体质量)]制备2型糖尿病大鼠模型。将Wistar大鼠50只随机分为正常对照组(NC组)10只和造模组40只,将造模成功的36只大鼠随机分为糖尿病组(DM组)、红芪多糖组(HPS组)、厄贝沙坦组(IRB组)及红芪多糖联合厄贝沙坦组(HPS+IRB组)各9只。HPS组每日予红芪多糖150 mg/kg灌胃,IRB组每日予厄贝沙坦30 mg/kg灌胃,HPS+IRB组每日予红芪多糖150 mg/kg加厄贝沙坦30 mg/kg灌胃,NC组和DM组予等量生理盐水灌胃,干预8周。检测各组大鼠空腹血糖,计算心重指数(H/B),实时荧光定量PCR法测定心肌局部转化生长因子β1(TGF-β1)、Smad3及Smad7 mRNA水平,免疫组化法测定TGF-β1、Smad3及Smad7蛋白水平,透射电镜下观察大鼠心肌超微结构,Masson染色法测定心肌胶原含量。结果:1)与NC组比较,其余4组大鼠心肌胶原含量增多,间质纤维化明显,H/B、TGF-β1、Smad3表达均升高,Smad7表达降低(P<0.01);与DM组相比,HPS组、IRB组及HPS+IRB组大鼠心肌纤维化减轻,H/B、TGF-β1、Smad3表达下降,Smad7表达升高(P<0.05),HPS+IRB组上述变化更显著。2)心肌病理改变:与NC组相比,DM组大鼠心肌胶原纤维增多,排列紊乱,线粒体肿胀;而HPS组、IRB组及HPS+IRB组上述改变减轻,HPS+IRB组上述改变减轻较HPS组和IRB组更明显。结论:红芪多糖及厄贝沙坦能通过抑制TGF-β1/Smads信号通路延缓2型糖尿病大鼠心肌纤维化的发生和发展,两药联合效果优于两药单用。

Objective:To discuss the effects of only hedysarum polybotys saccharide(HPS)or combined with irbesartan on myocardial fibrosis of type two diabetic rats.Methods:High-fat high-sugar fodder and one-off abdominal injection of streptozotocin(STZ,30 mg/kg,body mass)were used to prepare type two diabetes mellitus(T2DM)rat models.Fifty Wistar rats were randomized into ten rats in normal control group(NC group)and forty rats in the modelling group,after successfully modelling,36 rats were randomly allocated to DM group,HPS group,irbesartan(IRB)group and HPS+IRB group,nine rats each group.Each day,HPS group were drenched with HPS,150 mg/kg,IRB group with IRB,30 mg/kg,combination intervention group with HPS,150 mg/kg,and IRB,30 mg/kg,NC group and DM group equivalent dose of physiological saline,intervened for eight weeks.To detect fasting blood glucose and calculate heart weight index,real-time fluoresecence quantitative PCR was used to measure the levels of TGF-β1,Smad3 and Smad7 mRNA in myocardial areas,immunohistochemical method was adopted to measure the levels of TGF-β1,Smad3 and Smad7 protein,transmission electron microscope(TEM)was used to observe myocardial ultrastructure of the rats,Masson staining was used to detect the contents of myocardial collagen.Results:1)Compared with NC group,the contents of myocardial collagen increased in the rats of the other four groups,interstitial fibrosis was notable,H/B,the expressions of TGF-β1 and Smad3 lifted,the expressions of Smad7 lowered(P<0.01);compared with DM group,myocardial fibrosis relieved in HPS group,IRB group and HPS+IRB group,H/B,the expressions of TGF-β1 and Smad3 reduced,the expressions of Smad7 rose(P<0.05),these changes in HPS+IRB group were more notable.2)Myocardial pathological changes:Compared with NC group,myocardial collagen fiber in DM group increased,arranged in disorder,mitochondrial swelling;the above changes in HPS group,IRB group and HPS+IRB group relieved,the changes of HPS+IRB group were more obvious compared with HPS group and IRB group.Conclusion:HPS and irbesartan could delay the incidence and development of myocardial fibrosis of T2DM rats by inhibiting TGF-β1/Smads signal channel,the combination of both medicine are superior to single use of both.