甜菜苷抑制肾成纤维细胞增殖及转分化的作用及机制研究  

Study on effect and mechanism of betanin on inhibiting proliferation and transdifferentiation of renal fibroblasts

作　　者：何红叶 陈垦 成方玲 杨雨雪 金冉阳 邓智琴 王红勇 杨永健 He Hongye;Chen Ken;Cheng Fangling;Yang Yuxue;Jin Ranyang;Deng Zhiqin;Wang Hongyong;Yang Yongjian(College of Medicine,Southwest Jiaotong University,Chengdu,Sichuan,610031,China;Department of Cardiovascular Medicine,General Hospital of Western Theater Command of PLA,Chengdu,Sichuan,610083,China;Department of Cardiology,Daping hospital,Third Military Medical University,Chongqing,400042,China)

机构地区：[1]西南交通大学医学院,成都610031 [2]西部战区总医院心血管内科 [3]陆军军医大学大坪医院心血管内科

出　　处：《西南国防医药》2020年第3期224-229,共6页Medical Journal of National Defending Forces in Southwest China

基　　金：重庆市自然科学基金重点项目(cstc2015jcyjBX0129)。

摘　　要：目的探讨甜菜苷(Betanin)对血管紧张素Ⅱ(AngⅡ)诱导的肾成纤维细胞细胞增殖及转分化的作用及机制。方法原代培养小鼠肾脏成纤维细胞,将细胞分为溶剂对照组、AngⅡ处理组和AngⅡ+Betanin组。为探索Betanin对AngⅡ诱导的成纤维细胞影响的可能机制,在AngⅡ和Betanin处理的基础上,再将细胞分为溶剂对照组和Compound C组。采用细胞计数试剂盒(CCK-8)检测细胞增殖,实时定量聚合酶链式反应(qRT-PCR)、蛋白印迹(Western blotting)等方法检测增殖细胞核抗原(PCNA)、α-平滑肌肌动蛋白(α-SMA)、纤维连接蛋白Ⅰ(FN1)、Ⅰ型胶原蛋白(Col1)、腺苷酸活化蛋白激酶(AMPK)及其磷酸化表达变化。结果与溶剂对照组相比,AngⅡ组细胞增殖能力增加,PCNA、α-SMA、FN1、Col1表达增加(P<0.05)。与AngⅡ组比较,Betanin能抑制细胞增殖,减少AngⅡ诱导的PCNA、α-SMA、FN1、Col1表达(P<0.05),同时上调AMPK磷酸化水平(P<0.05)。加入AMPK抑制剂Compound C阻遏了Betanin的上述作用(P<0.05)。结论Betanin可通过激活AMPK,拮抗AngⅡ诱导的肾间质成纤维细胞增殖、转分化以及细胞外基质产生,提示Betanin在高血压肾纤维化中具有良好的应用前景,或能成为延缓向终末期肾病进展的治疗手段。Objective To explore the effect and mechanism of betanin on inhibiting cell proliferation and transdifferentiation of renal fibroblasts induced by angiotensinⅡ(AngⅡ).Methods The primary cultured renal fibroblasts of mice were divided into the vehicle control group,AngⅡtreatment group and AngⅡ+Betanin group.In the AngⅡ+Betanin group,the cells were subdivided into the vehicle control subgroup and Compound C subgroup to explore the possible mechanism of Betanin in the AngⅡ-induced effect on fibroblasts.Cell Counting Kit-8(CCK-8)was used to test the cell proliferation,and quantitative real-time polymerase chain reaction(qRT-PCR)and western blotting were used to test the changes of expressions of proliferation cell nuclear antigen(PCNA),α-smooth muscle actin(α-SMA),fibronectinⅠ(FN1),collagenⅠ(Col1),adenosine monophosphate-activated protein kinase(AMPK)and AMPK phosphorylation.Results Compared with the vehicle control group,the cell proliferative ability of AngⅡgroup was enhanced,and the expressions of PCNA,α-SMA,FN1 and Col1 were increased(P<0.05).Compared with AngⅡgroup,Betanin in the AngⅡ+Betanin group could inhibit the cell proliferation,reduce the expressions of AngⅡ-induced PCNA,α-SMA,FN1 and Col1(P<0.05)and increase the level of AMPK phosphorylation(P<0.05).Addition of Compound C,an inhibitor of AMPK,suppressed the abovementioned effects of Betanin(P<0.05).Conclusion With activation of AMPK,Betanin can inhibit the AngⅡ-induced proliferation,transdifferentiation and production of extracellular matrix of renal interstitial fibroblasts,which suggests that Betanin is promising to be applied in hypertensive renal fibrosis and may be a treatment to delay the progression to end-stage renal disease.

关 键 词：甜菜苷 血管紧张素Ⅱ 肾成纤维细胞 腺苷酸活化蛋白激酶 

分 类 号：R285.5[医药卫生—中药学]