中药心康方对大鼠心力衰竭模型心肌胶原代谢的影响

Effects of Xinkang recipe on myocardial collagen metabolism in rats with doxorubicin-induced heart failure

目的观察中药心康方对大鼠心力衰竭模型心肌胶原代谢的影响。方法阿霉素腹腔注射法建立大鼠心力衰竭模型,随机分为对照组(10只)、模型组(9只)、心康方组(9只)和卡托普利组(9只)。Masson染色观察心肌胶原变化,Western blot检测心肌Ⅰ型、Ⅲ型胶原、转化生长因子β1(TGF-β1)、核因子κB的抑制蛋白(IκB)和p56的表达,Western blot和RT-q PCR分别检测心肌基质金属蛋白酶(MMP)-2、MMP-9、基质金属蛋白酶组织抑制因子(TIMP)-1和TIMP-2的蛋白和mRNA表达水平。结果与对照组比较,模型组大鼠心肌Ⅰ型和Ⅲ型胶原表达显著增加,胶原容积分数显著升高(均为P<0.01);TGF-β1和p56表达显著增加,IκB显著降低(均为P<0.05);MMP-2、MMP-9、TIMP-1和TIMP-2的蛋白和mRNA表达水平显著升高(P<0.01或P<0.05)。与模型组比较,心康方组和卡托普利组大鼠的心肌Ⅰ型和Ⅲ型胶原表达显著减少,胶原容积分数显著降低(均为P<0.01);TGF-β1和p56显著降低,IκB显著升高(均为P<0.05);MMP-2、MMP-9、TIMP-1和TIMP-2的蛋白和mRNA表达水平显著降低(P<0.01或P<0.05)。结论中药心康方可减轻心力衰竭大鼠的心肌胶原沉积,其机制可能与抑制核因子κB介导的TGF-β1上调有关。

Objective To investigate the effects of Xinkang recipe on myocardial collagen metabolism in rats with doxorubicin-induced heart failure. Methods The heart failure rat model was induced by intraperitoneal injection of doxorubicin. Male SD rats were randomly divided into the control group （n =10）,the model group（ n = 9） , the Xinkang group（ n = 9 ） and the captopril group（ n =10）. Masson＇s trichrome staining was used to observe myocardial fibrosis. Western blotting was used to detect of expression of collagen I ,collagen Ⅲ, TGF-p1, IkB and p56 in myocardium. Myocardial protein and mRNA levels of matrix metalloproteinase-2 （ MMP-2 ） , MMP-9, tissue inhibitor of matrix metalloproteinase-1 （TIMP-1 ） and TIMP-2 were detected by Western blotting and RT-qPCR, respectively. Results Compared with control group,expression of type I and type Ⅲ collagen protein significantly increased, and the collagen volume fraction（ CVF） significantly increased in the model group（all P 〈0. 0 1 ） . TGF-p1 and p56 protein expression significantly increased,while IkB protein expression significantly decreased in the model group （a l l P 〈 0. 05） . The myocardial protein and mRNA levels of MMP-2, MMP-9, TIMP-1 and TIMP-2 were significantly higher in the model group（ P 〈0. 01 or P 〈0. 05） . Compared with the model group, expression of collagen I and collagen Ⅲ protein significantly decreased,and the CVF significantly reduced in Xinkang group and captopril group（both P 〈 0. 01）. The TGF-β1 and p56 protein expression significantly decreased, and the IkB protein expression significantly increased in Xinkang group and captopril group （all P 〈 0 . 0 5 ） . The levels of protein and mRNA of MMP-2, MMP-9, TIMP-1 and TIMP-2 were significantly lower in Xinkang group and captopril group （ P 〈0. 01 or P 〈0. 05 ） . Conclusions Xinkang recipe can reduce collagen deposition in myocardium in rats with doxorubicin-induced heart failure. Down-regulating of TGF-β1 protein expression through the inhibition of NF-kB signal transduction pathway might be a potential mechanism underlying effects of Xinkang recipe.