漆黄素对血管紧张素Ⅱ诱导心肌成纤维细胞活性的影响

Effect of urushiflavin on viability of angiotensinⅡ-induced myocardial fibroblasts

目的探讨漆黄素对血管紧张素Ⅱ(AngⅡ)诱导心肌成纤维细胞活性及白细胞介素33(IL-33)/白细胞介素1受体样分子1(ST2)信号通路的影响。方法体外培养人心肌成纤维细胞,并将细胞分为对照组、AngⅡ组、漆黄素1组(1.25μmol/L)、漆黄素2组(2.5μmol/L)、漆黄素3组(5.0μmol/L)。细胞计数试剂盒8法检测心肌成纤维细胞活力。免疫荧光染色检测α平滑肌肌动蛋白(α-SMA)表达。ELISA法检测Ⅰ型胶原、Ⅲ型胶原表达。Western blot法检测增殖蛋白β连环蛋白、细胞周期蛋白D1及α-SMA、IL33/ST2信号通路蛋白表达。结果与对照组比较,AngⅡ组心肌成纤维细胞活力、α-SMA、Ⅰ型胶原、Ⅲ型胶原、β连环蛋白、细胞周期蛋白D1、IL-33、ST2表达明显升高,差异有统计学意义(P<0.05)。与AngⅡ组比较,漆黄素1组、漆黄素2组、漆黄素3组心肌成纤维细胞活力、α-SMA、Ⅰ型胶原、Ⅲ型胶原、β连环蛋白、细胞周期蛋白D1表达明显降低,且随着漆黄素浓度升高,心肌成纤维细胞各指标呈剂量依赖性降低,差异有统计学意义(P<0.05)。与AngⅡ组比较,漆黄素1组、漆黄素2组、漆黄素3组心肌成纤维细胞IL-33、ST2表达明显降低(1.26±0.21、0.77±0.15、0.29±0.13 vs 1.75±0.19,1.15±0.14、0.64±0.12、0.28±0.09 vs 1.62±0.16,P<0.05),且随着漆黄素浓度升高,IL-33、ST2表达逐渐降低,差异有统计学意义(P<0.05)。结论漆黄素能够抑制AngⅡ诱导的心肌成纤维细胞活化及IL-33/ST2信号通路,可能是心肌纤维化治疗的潜在药物。

Objective To study the effects of urushiflavin on viability of angiotensinⅡ(AngⅡ)-induced myocardial fibroblast,interleukin(IL)-33/IL receptor-like molecule 1(ST2)signaling pathway.Methods Human myocardial fibroblasts cultured in vitro were divided into control group,AngⅡgroup and urushiflavin(1.25μmol/L)group 1,urushiflavin(2.5μmol/L)group 2 and urushiflavin(5.0μmol/L)group 3.The viability of myocardial fibroblasts was tested with cell counting kit 8 in different groups.The expressions ofα-smooth muscle actin(α-SMA)were detected with immunofluoresence staining,those of collagen typeⅠandⅢwere detected by enzyme-linked immunosorbent assay,and those of cell proliferation-related proteinsβ-catenin,cyclin-D1 and IL-33/ST2 signaling pathway proteins were detected by Western blot.Results The viability of myocardial fibroblasts,expression levels ofα-SMA,collagen typeⅠ,collagen typeⅢ,β-catenin,cyclin-D1,IL-33/ST2 signalling pathway proteins were significantly higher in AngⅡgroup than in control group(P<0.05)and significantly lower in urushiflavin groups 1-3 than in AngⅡgroup(P<0.05),and decreased in a dose-dependent manner with the increasing serum urushiflavin level(P<0.05).The expression levels of IL-33 and ST2 in myocardial fibroblasts were significantly lower in urushiflavin groups 1-3 than in AngⅡgroup(1.26±0.21,0.77±0.15,0.29±0.13 vs 1.75±0.19;1.15±0.14,0.64±0.12,0.28±0.09 vs 1.62±0.16,P<0.05)and decreased gradually in a dose-dependent manner with the increasing serum urushiflavin level(P<0.05).Conclusion Urushiflavin can inhibit AngⅡ-induced viability of myocardial fibroblasts and IL-33/ST2 signaling pathway,and can thus be used as a potential drug in treatment of myocardial fibrosis.