人参皂苷Re调控Wnt/β-catenin信号通路抑制心肌成纤维细胞增殖研究  

Research on Ginsenoside Re inhibition of CFs proliferation by regulating Wnt/p-catenin signaling pathway

作　　者：韩路拓 郭宏伟 任钧国[3] 刘建勋[3] 王攀 杨佳妹 Han Lutuo;Guo Hongwei;Ren Junguo;Liu Jianxun;Wang Pan;Yang Jiamei(Department of Chinese Medicine Museum,Heilongjiang University of Chinese Medicine,Harbin 150040,China;department of Basic Medical College,Heilongjiang University of Chinese Medicine,Harbin 150040,China;institute of Basic Medical Sciences of Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing Key Laboratory of Pharmacology of Chinese Materia Medica,Beijing 100091,China)

机构地区：[1]黑龙江中医药大学中医药博物馆,哈尔滨150040 [2]黑龙江中医药大学基础医学院,哈尔滨150040 [3]中国中医科学院西苑医院基础医学研究所中药药理北京市重点实验室,北京100091

出　　处：《国际中医中药杂志》2022年第3期298-304,共7页International Journal of Traditional Chinese Medicine

基　　金：国家科技重大专项“重大新药创制”项目(2017ZX09301018);黑龙江省经济社会发展重点研究课题(20415)。

摘　　要：目的观察人参皂苷Re对体外高糖培养的原代心肌成纤维细胞(CFs)增殖与蛋白分泌的影响, 以及对Wnt/β-连接蛋白(β-catenin)信号通路的调控作用。方法采用体外高糖诱导CFs增殖模型, 经不同浓度的人参皂苷Re干预后, 采用MTT法检测CFs细胞增殖能力, 流式细胞仪检测细胞增殖周期, ELISA法检测Ⅰ型胶原、Ⅲ型胶原及TGF-β1水平, Western blot检测β-catenin、糖原合成酶激酶3β(GSK-3β)、磷酸化糖原合成酶激酶3β(p-GSK-3β)蛋白表达情况。结果与模型对照组比较, 人参皂苷Re各剂量组吸光度值降低(P<0.01), G0+G1期细胞百分比增加(P<0.01)、S+G2+M期细胞百分比降低(P<0.01), TGF-β1水平降低(P<0.01), 人参皂苷Re高、中剂量组Ⅲ型胶原[(6.566±1.620)ng/ml、(7.170±0.470)ng/ml比(11.241±2.234)ng/ml]水平降低(P<0.01), 人参皂苷Re高、中剂量组β-catenin[(0.281±0.016)、(0.301±0.021)比(0.409±0.037)]表达降低(P<0.01), 人参皂苷Re高剂量组p-GSK-3β[(0.369±0.049)比(0.268±0.048)]表达升高(P<0.01)。结论人参皂苷Re可调控Wnt/β-catenin 信号转导通路异常表达, 有效抑制CFs增殖, 减少高糖条件下CFs胶原和TGF-β1合成, 具有延缓糖尿病心肌纤维化的作用。Objective To observe the effect of Ginsenoside Re on the proliferation and protein secretion of primary cardiac fibroblasts(CFs)cultured in high glucose by vitro,and the regulation of Wnt/β-catenin signaling pathway.Methods The myocardial fibroblast proliferation model induced by high glucose in vitro was used.Cell proliferation was detected by MTT method,cell cycle was measured by flow cytometry,concentration of typeⅠ,Ⅲcollagens and TGF-β1 protein were tested by ELISA assay.Protein expression ofβ-catenin,GSK-3βand p-GSK-3βwere determined by Western blot.Results Compared with the model group,the cell proliferation in Ginsenoside Re high,medium,low group were significantly decreased(P<0.01),the percentage of cells in G 0+G 1 phase was increased(P<0.01),and the percentage of cells in S+G 2+M phase was decreased(P<0.01),the content of TGF-β1 was significantly decreased(P<0.01).The content of typeⅢcollagen[(6.566±1.620)ng/ml,(7.170±0.470)ng/ml vs.(11.241±2.234)ng/ml]in Ginsenoside Re high,medium group were significantly decreased(P<0.01).The expression ofβ-catenin(0.281±0.016,301±0.021 vs.0.409±0.037)was significantly decreased and the expression of p-GSK-3β(0.369±0.049 vs.0.268±0.048)in Ginsenoside Re high,medium group were significantly increased(P<0.01).Conclusion Ginsenoside Re plays an important role in inhibiting CFs proliferation and reduct the synthesis of collagn and TGF-β1 by regulating abnormal expression of Wnt/β-catenin signaling pathway.It has the potential to delay the myocardil fibrosis of diabetes mellitus.

关 键 词：人参皂苷RE WNT信号通路 细胞增殖 转化生长因子Β1 成纤维细胞 

分 类 号：R285[医药卫生—中药学]