防己茯苓汤对心肌纤维化小鼠巨噬细胞极化和氧化应激的影响

Effect of Fangji Fulingtang on Macrophage Polarization and Oxidative Stress in Mouse Model of Myocardial Fibrosis

目的:探讨防己茯苓汤对心肌纤维化小鼠巨噬细胞极化和氧化应激的影响。方法:通过皮下注射异丙肾上腺素(5 mg·kg^(-1)·d^(-1))构建心肌纤维化小鼠模型。将50只C57BL/6J小鼠随机分为假手术组、模型组、防己茯苓汤低、高剂量组、酒石酸美托洛尔(Meto)组,每组10只。假手术组、模型给予相应体积生理盐水灌胃,防己茯苓汤低、高剂量组分别予以防己茯苓汤低、高剂量灌胃(3.315、13.26 g·kg^(-1)·d^(-1)),Meto组予以酒石酸Meto灌胃(15 mg·kg^(-1)·d^(-1))。2周后,观察小鼠心脏形态,计算平均细胞横截面积、心脏质量指数和心胫比,组织病理学方法观察心肌胶原沉积,酶联免疫吸附测定法(ELISA)检测血清肌酸激酶同工酶(CK-MB)、转化生长因子-β_(1)(TGF-β_(1))、炎症因子指标[肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6、IL-10]和氧化应激指标[丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)]的表达水平,免疫组织化学染色观察心肌组织CD86和CD206蛋白表达水平。结果:与模型组比较,防己茯苓汤低、高剂量组和Meto组小鼠心脏外观形态明显减小,平均心肌细胞横截面积、心脏质量指数和心胫比显著降低(P<0.01);血清CK-MB、TGF-β1水平明显降低(P<0.05,P<0.01);血清促炎细胞因子TNF-α、IL-1β、IL-6表达水平明显降低(P<0.05,P<0.01),抗炎细胞因子IL-10明显升高(P<0.05);炎症细胞浸润、心肌损伤、胶原沉积和纤维化程度明显减轻;巨噬细胞极化标志物CD86表达水平显著降低(P<0.01),CD206表达水平显著升高(P<0.01);氧化应激指标MDA表达水平显著升高(P<0.01),SOD、GSH表达水平显著降低(P<0.01)。结论:防己茯苓汤可以改善异丙肾上腺素诱导的心肌纤维化,其机制可能是通过调控巨噬细胞极化和氧化应激发挥作用。

Objective:To investigate the effects of Fangji Fulingtang on macrophage polarization and oxidative stress in the mouse model of myocardial fibrosis.Method:The mouse model of myocardial fibrosis was established by subcutaneous injection of isoproterenol(ISO,5 mg·kg^(-1)·d^(-1)).Fifty C57BL/6J mice were randomly assigned into control(0.9%NaCl),model(0.9%NaCl),low-and high-dose(3.315 g·kg^(-1)·d^(-1) and 13.26 g·kg^(-1)·d^(-1),respectively)Fangji Fulingtang(FFD-L and FFD-H,respectively),and metoprolol tartrate(Meto,15 mg·kg^(-1)·d^(-1))groups,with 10 mice each group.After 2 weeks of treatment,the heart appearance,cardiac weight index(CWI),heart weight(HW)/tibia length(TL)ratio,and myocardial histopathological alterations were observed.Meanwhile,the serum levels of creatine kinase-MB(CK-MB),transforming growth factor-β_(1)(TGF-β_(1)),tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-6,IL-10,malondialdehyde(MDA),superoxide dismutase(SOD),and glutathione(GSH)were measured by enzyme-linked immunosorbent assay(ELISA).The expression levels of CD86 and CD206 were observed by immunohistochemical staining.Result:Compared with the model group,the FFD-L,FFD-H,and Meto groups showed improved heart appearance,decreased CWI and HW/TL ratio(P<0.01),lowered serum levels of CK-MB,TGF-β1,TNF-α,IL-1β,and IL-6(P<0.05,P<0.01),and elevated IL-10 level(P<0.05).Furthermore,the three groups showed reduced infiltration of inflammatory cells,myocardial injury,collagen deposition,and myocardial fibrosis,decreased CD86,SOD,and GSH(P<0.01),and increased CD206 and MDA(P<0.01).Conclusion:Fangji Fulingtang can mitigate ISO-induced myocardial fibrosis by regulating macrophage polarization and oxidative stress.