Smad3、TGF-β1蛋白检测在心肌肥厚大鼠体内的表达

Expression of Smad3 and TGF-β1 protein in rats with cardiac hypertrophy

目的 探讨免疫组化法检测Smad3蛋白(Smad3)、转化生长因子β1 (Transforming growth factor betal,TGF-β1)蛋白在肥厚型心肌病(Hypertrophic cardiomyopathy,HCM)大鼠体内的表达概况。方法 选择SPF级SD新生大鼠36只,随机分为4组,A组、B组(建模后3 d)、C组(建模后7 d)及D组(建模后14 d),各9只。B-D组建立HCM大鼠模型,A组仅分离腹主联率接扎,各组颈部脱白前检测左心室重量指数,处死后,分别采用苏木精HE检测病理,免疫组化/印迹及PCR检测Smad3、TGF-β1表达。结果 建模后3 d、7 d及14 d的B~D组大鼠直请LVMI水平均高于A组,建模14d后LVMI水平达到峰值;A组心肌组织整齐,纹理清晰,B~D组心肌组织逐渐出现水肿坏死,心肌纤维扩张及肌丝溶解,D组心肌组织空泡变性,细胞核国缩严重,大量炎症浸润。B组~D组大鼠心肌组织中TGF BI/Smad3阳性表达均高于A组,各组相比差异明显(P<0.001):B组~D组大鼠心肌组织中TCF-B1/Smad3蛋白相对表达均高于A组,各组相比差异明显(P<0001):B组~D组大鼠心肌组织中TGF-Bl/Sad3mRNA水平高于A组,各组相比差异明显(P<0.001)。结论 TGF-B1/Smad3能够诱导心肌组织坏死及水肿,随着病情的加重TGF-B1Sma3水随之增加,说明TGF-B1Smd3参与HCM病理过程。

Objective To investigate the detection of Smad3 protein and Transforming growth factor betal by immunohistochemical method.Expression of TGF-β1 protein in Hypertrophic cardiomyopathy(HCM)rats.Methods Thirty-six SPF SD neonatal rats were randomly divided into 4 groups:group A,group B(3 days after modeling),group C(7 days after modeling)and group D(14 days after modeling),9 rats in each group.In group A,the left ventricular mass index was detected before neck decalescence.After the rats were sacrificed,hematoxylin HE was used to detect the pathology,immunohistochemistry/Western blotting and PCR were used to detect the expression of Smad3 and TGF-β1.Results The LVMI level in group B to D was higher than that in group A at 3 d,7d and 14d after modeling,and the LVMI level reached the peak at 14d after modeling.The myocardial tissue of group A was neat and clear in texture.The myocardial tissue of groups B to D gradually showed edema and necrosis,myocardial fiber expansion and myofilaments lysis.The myocardial tissue of group D had vacuolar degeneration,severe nuclear shrinkage and massive inflammatory infiltration.The positive expression of TGF BI/Smad3 in myocardial tissue of rats in groups B to D was higher than that in group A,and the difference was significant(P<0.001).The relative expression of TCF-B1/Smad3 protein in myocardial tissue of rats in groups B to D was higher than that in group A,and the difference was significant(P<0.001):The mrna level of TGF-Bl/SAD3 in myocardial tissue of rats in groups B to D was higher than that in group A,and the difference was significant(P<0.001).Conclusion TGF-B1/Smad3 can induce myocardial tissue necrosis and edema,and TGF-B1Sma3 water increases with the aggravation of the disease,indicating that TGF-B1Smd3 is involved in the pathological process of HCM.