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目的:探讨苓桂术甘汤(LGZGD)含药血清对转化生长因子-β1(transforming growth factor-β1,TGF-β1)诱导心肌纤维化的保护作用。方法:利用酶消化,结合差速贴壁的方法体外分离及培养乳鼠的心肌成纤维细胞(cardiac fibroblasys,CFB)。分为:空白组、20%空白大鼠血清组、模型组、5%LGZGD含药血清组、10%LGZGD含药血清组和20%LGZGD含药血清组,除空白组和空白大鼠血清组外分别予5 ng/mL TGF-β1刺激12 h,再用LGZGD含药血清(5%、10%、20%)进行干预后继续培养24 h。采用免疫荧光法及蛋白质印迹法(western blot,WB)检测α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的表达,酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)及WB检测I型胶原蛋白(Collagen I)、Ⅲ型胶原蛋白(CollagenⅢ)及纤维连接蛋白(fibronectin,FN)的表达。结果:与空白组比较,模型组Collagen I、CollagenⅢ、α-SMA和FN的表达均显著升高(P<0.01),与模型组比较,实验各浓度组Collagen I及CollagenⅢ的表达均显著降低(P<0.01);α-SMA和FN的表达显著降低(P<0.01)。结论:LGZGD对胶原蛋白合成以及α-SMA和FN的表达有抑制作用,表明LGZGD的抗纤维化作用与其有关。
Abstract：Objective:To explore the protective effect of Linggui Zhugan Decoction(LGZGD)medicated serum on myocardial fibrosis induced by transforming growth factor-β1(TGF-β1).Methods:Using enzyme digestion method,combined with differential adherence to isolate and culture suckling mouse cardiac fibroblasys(CFB)in vitro. Divided into:blank group,blank rat serum group,model group,and LGZGD medicated serum group(5%,10%,20%). Except for blank group and blank rat serum group,they were stimulated with 5 ng/mL TGF-β1 for 12 h,and then the intervene with LGZGD medicated serum(5%,10%,20%)and continue to culture for 24 h. Use immunofluorescence and western blot(WB)to detect the expression of α-smooth muscle actin(α-SMA),enzyme-linked immunosorbent assay(ELISA)and WB to detect type I collagen(Collagen I),type I collagen(Collagen Ⅲ)and fibronectin(FN)expression.Results:Compared with the blank group,the expressions of Collagen I,Collagen Ⅲ,α-SMA and FN in the model group were significantly increased(P<0.01);Compared with the model group,the expressions of Collagen I and Collagen Ⅲ in each concentration group of the experiment were significantly reduced(P<0.01);the expression of α-SMA and FN were significantly reduced(P<0.01).Conclusions:LGZGD has an inhibitory effect on collagen synthesis and the expression of α-SMA and FN,indicating that the anti-fibrosis effect of LGZGD is related to it.
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基本信息:
DOI：10.13210/j.cnki.jhmu.20201229.002
中图分类号:R285.5
引用信息:
[1]葛瑞瑞,王翔,汤同娟,等.苓桂术甘汤含药血清对TGF-β_1诱导的心肌成纤维细胞α-SMA和胶原蛋白合成的影响[J].海南医学院学报,2021,27(10):741-746+751.DOI:10.13210/j.cnki.jhmu.20201229.002.
基金信息:
国家自然科学基金资助项目（81973844）~~
2020-12-29
2020-12-29
2020-12-29