抗纤益心方调节MCP-1/CCR2信号通路对扩张型心肌病大鼠心功能的影响

Effects of Kangxian Yixin Formula on cardiac function of rats with dilated cardiomyopathy via regulating MCP-1/CCR2 signaling pathway

目的:探讨抗纤益心方调节单核细胞趋化蛋白-1(monocyte chemotactic protein-1,MCP-1)/趋化因子C-C-基元受体2(C-C-subunit receptor 2,CCR2)信号通路对扩张型心肌病(dilated cardiomyopathy,DCM)大鼠心功能的影响。方法:取SD雄性大鼠随机分为对照组、模型组、抗纤益心方(18.8 g·kg^(-1))组、抗纤益心方(18.8 g·kg^(-1))+空载(空载质粒)组、抗纤益心方(18.8 g·kg^(-1))+MCP-1过表达(MCP-1过表达质粒)组,模型组和给药干预组大鼠以腹腔注射阿霉素法诱导建立DCM模型,对照组大鼠于腹腔内注射等剂量生理盐水,以抗纤益心方(每天1次)和质粒(每周2次)分组干预3周后检测各组大鼠心功能指标左室舒张末期内径(left ventricular end diastolic diameter,LVEDD)、左室收缩末期内径(left ventricular end systolic diam⁃eter,LVESD)、左心射血分数(left ventricular ejection fraction,LVEF)、左室短轴缩短率(left ventricular short axis shortening rate,FS)、心脏指数、心肌组织病理形态及纤维化、血清肿瘤坏死因子α(TNF-α)、转化生长因子β1(TGF-β1)及MCP-1水平、心肌组织胶原[Ⅰ型胶原蛋白(Col-Ⅰ)、Ⅲ型胶原蛋白(Col-Ⅲ)、α平滑肌肌动蛋白(α-SMA)]生成及MCP-1/CCR2通路相关蛋白表达。结果:与对照组相比,模型组大鼠心肌组织发生严重损伤,心脏指数、LVEDD、LVESD、胶原容积分数(CVF)、血清TNF-α、TGF-β1及MCP-1水平,心肌组织Col-Ⅰ、Col-Ⅲ、α-SMA、MCP-1、CCR2蛋白表达显著升高(P<0.05),LVEF、FS显著降低(P<0.05)。与模型组比较,抗纤益心方组大鼠心肌组织损伤减轻,心脏指数、LVEDD、LVESD、CVF、血清TNF-α、TGF-β1及MCP-1水平,心肌组织Col-Ⅰ、Col-Ⅲ、α-SMA、MCP-1、CCR2蛋白表达降低(P<0.05),LVEF、FS升高(P<0.05);抗纤益心方+空载组大鼠各指标无明显变化(P>0.05)。过表达MCP-1可减弱抗纤益心方对DCM大鼠各指标的影响。结论:抗纤益心方可通过降低MCP-1/CCR2信号通路活性而抑制DCM大鼠炎症,进而减轻大鼠心肌损伤及纤维化,改善其心功能。

OBJECTIVE To investigate the effects of Kangxian Yixin Formula on cardiac function of rats with dilated cardiomyopathy(DCM)by modulating monocyte chemotactic protein-1(MCP-1)/chemokine C-C-subunit receptor 2(CCR2)signaling pathway.METHODS SD male rats were randomly divided into five groups:control group,model group,Kangxian Yixin Formula(18.8 g·kg^(-1))group,Kangxian Yixin Formula(18.8 g·kg^(-1))+empty(empty plasmid)group,and Kangxian Yixin Formula(18.8 g·kg^(-1))+MCP-1 overexpression(MCP-1 overexpression plasmid)group.The model group and intervention groups were induced to develop a DCM model by intraperitoneal injection of doxorubicin,while the control group received an equal dose of saline intraperitoneally.After 3 weeks of daily intervention with Kangxian Yixin Formula and twice-a-week plasmid intervention,cardiac function parameters were measured,including left ventricular end diastolic diameter(LVEDD),left ventricular end systolic diameter(LVESD),left ventricular ejection fraction(LVEF),left ventricular short axis shortening rate(FS),heart index,myocardial tissue pathology and fibrosis,serum levels of tumor necrosis factor-α(TNF-α),transforming growth factor-β1(TGF-β1),and MCP-1,as well as the expression of collagen production(typeⅠcollagen(Col-Ⅰ),typeⅢcollagen(Col-Ⅲ),α-smooth muscle actin(α-SMA)and MCP-1/CCR2 pathway related proteins in myocardial tissue of rats in each group.RESULTS Compared with the control group,the model group exhibited severe myocardial tissue damage,and significant increase in heart index,LVEDD,LVESD,collagen volume fraction(CVF),serum levels of TNF-α,TGF-β1,and MCP-1,and expression of myocardial tissue Col-Ⅰ,Col-Ⅲ,α-SMA,MCP-1,and CCR2 proteins(P<0.05),along with a significant decrease in LVEF and FS(P<0.05).Compared with the model group,the Kangxian Yixin Formula group showed reduced myocardial tissue damage,decreased cardiac index,LVEDD,LVESD,CVF,serum TNF-α,TGF-β1,and MCP-1,and expression of myocardial tissue Col-Ⅰ,Col-Ⅲ,α-SMA,MCP-1,and CCR2 proteins(P<0.05),with an increase in LVEF and FS(P<0.05).There was no obvious change in any indicators in the Kangxian Yixin Formula+empty group(P>0.05).Overexpression of MCP-1 could weaken the effects of Kangxian Yixin Formula on various indicators in DCM rats.CONCLUSION Kangxian Yixin Formula can inhibit inflammation in DCM rats by reducing the activity of MCP-1/CCR2 signaling pathway,thereby alleviating myocardial injury and fibrosis in rats and improving their cardiac function.