益肺汤联合MitoQ对TGF-β1诱导的人肺泡上皮细胞HPAEpiC间质转化的抑制作用  

Inhibitory effect of Yifei decoction combined with MitoQ on interstitial transformation of human alveolar epithelial cells HPAEpiC induced by TGF-β1

作　　者：陈丽娟[1] 肖泓[1] 兰成仲 杨彦斌 祁向荣 欧阳丽[1] 张晓丽 王凤英 金其花 孙毅 CHEN Lijuan;XIAO Hong;LAN Chengzhong;YANG Yanbin;QI Xiangrong;OUYANG Li;ZHANG Xiaoli;WANG Fengying;JIN Qihua;SUN Yi(Department of Pulmonology,First Affiliated Hospital of Yunnan College of Traditional Chinese Medicine,Yunnan Traditional Chinese Medicine Hospital,Kunming 650000,China;Department of Urology,First Affiliated Hospital of Yunnan College of Traditional Chinese Medicine,Yunnan Traditional Chinese Medicine Hospital)

机构地区：[1]云南中医学院第一附属医院,云南省中医医院肺病科,昆明650000 [2]云南中医学院第一附属医院,云南省中医医院泌尿科

出　　处：《山西医科大学学报》2021年第3期270-276,共7页Journal of Shanxi Medical University

基　　金：云南省科技厅中医联合青年项目(2018FF001(-080));云南省科技厅中医联合面上项目(2019FF002(-034))。

摘　　要：目的探讨益肺汤联合线粒体辅酶Q(MitoQ)对生长转化因子(TGF-β1)诱导的人肺泡上皮细胞HPAEpiC间质转化的抑制作用。方法选择细胞株HPAEpiC为研究对象,采用TGF-β1(10 ng/ml)诱导的上皮间质转化模型进行实验。设置空白对照组、模型组、模型+MitoQ组、模型+益肺汤含药血清组、模型+MitoQ+益肺汤含药血清组。采用细胞计数试剂盒8(CCK-8)检测益肺汤联合MitoQ对细胞存活率的影响。实时荧光定量(qRT-PCR)分析益肺汤联合MitoQ对细胞内E-cadherin、ColⅠ、α-SMA mRNA表达水平的影响,流式细胞术检测细胞内ROS水平变化,蛋白质印迹法(Western blot)检测细胞内GRP78蛋白表达水平。结果与空白对照组比较,模型组细胞存活率明显增加,细胞ColⅠ、α-SMA mRNA表达、GRP78蛋白表达及ROS荧光强度明显增加,细胞E-cadherin mRNA表达明显降低(P<0.05);与模型组比较,益肺汤联合MitoQ组细胞内ColⅠ、α-SMA mRNA表达有效降低(P<0.05),E-cadherin mRNA表达增加(P<0.01),细胞内GRP78蛋白表达及ROS荧光强度降低(P<0.05)。结论益肺汤联合MitoQ对TGF-β1诱导的人肺泡上皮细胞HPAEpiC上皮间质转化有一定的抑制作用,为肺纤维化潜在的治疗药物,其可能的作用机制为下调ColⅠ、α-SMA mRNA及GRP78蛋白表达,上调E-cadherin mRNA表达。Objective To investigate the inhibitory effect of Yifei decoction combined with mitoquinone(MitoQ)on the interstitial transformation of human alveolar epithelial cells HPAEpiC induced by TGF-β1.Methods The cell line HPAEpiC was cultured and divided into control group,model group,model+MitoQ group,model+Yifei decoction group,and model+MitoQ+Yifei decoction group.The model of epithelial-mesenchymal transformation was induced by 10 ng/ml TGF-β1 for the experiment.The cell viability was detected by CCK-8,the mRNA expression of E-cadherin,ColⅠandα-SMA was analyzed by qRT-PCR,the level of intracellular ROS was detected by flow cytometry,and the expression of GRP78 protein was detected by Western blot.Results Compared with control group,the cell survival rate,the expression levels of ColⅠ,α-SMA mRNA,GRP78 protein and ROS fluorescence intensity were significantly increased in model group(P<0.05),while the expression of E-cadherin mRNA was significantly decreased(P<0.05).Compared with model group,the expression of ColⅠandα-SMA mRNA was reduced in model+MitoQ+Yifei decoction group(P<0.05),the expression of E-cadherin mRNA was increased(P<0.01),and the expression of GRP78 protein and ROS fluorescence intensity in cells were decreased(P<0.05).Conclusion Yifei decoction combined with MitoQ can inhibit the epithelial-mesenchymal transformation of human alveolar epithelial cells HPAEpiC induced by TGF-β1 by down-regulating the expression of ColⅠ,α-SMA mRNA and GRP78 protein and up-regulating the expression of E-cadherin mRNA.It can be a potential therapeutic drug for pulmonary fibrosis.

关 键 词：益肺汤 人肺泡上皮细胞 线粒体辅酶Q 间质转化 

分 类 号：R563[医药卫生—呼吸系统]