发酵黄芪对人胚肺二倍体成纤维细胞增殖及衰老相关蛋白表达的影响  

Effects of fermented milkvetch root on proliferation and expression of aging-related proteins in human embryonic lung diploid fibroblast cell

作　　者：吴爽[1] 李学锋[2] 王秋彤 李喆[1] 刘清新[1] WU Shuang;LI Xuefeng;WANG Qiutong;LI Zhe;LIU Qingxin(Cangzhou Medical College,Cangzhou 061000,China;Department of General Surgery,Cangzhou Central Hospital,Cangzhou 061000,China)

机构地区：[1]沧州医学高等专科学校,河北沧州061000 [2]沧州市中心医院普外科,河北沧州061000

出　　处：《长春中医药大学学报》2020年第4期676-679,共4页Journal of Changchun University of Chinese Medicine

基　　金：沧州市重点研发计划指导项目(183302104)。

摘　　要：目的探讨发酵黄芪对人胚肺二倍体成纤维细胞增殖及衰老相关蛋白表达的影响。方法体外培养人胚肺二倍体成纤维细胞(HDF),并根据代龄和加药情况分为青年组(30代前的HDF细胞),模型组(40代后的HDF细胞),对照组(用10%正常血清处理的40代后的HDF细胞),实验组(用10%发酵黄芪干预血清处理的40代后HDF细胞)。培养至40代时开始干预,光学倒置显微镜观察各组HDF细胞形态;采用活细胞计数(CCK-8)法检测各组HDF细胞增殖情况;采用分光光度法检测各组HDF细胞β-半乳糖苷酶(β-gal)活性;采用蛋白免疫印迹(Wb)法检测各组HDF细胞p16、细胞周期蛋白D1(CyclinD1)、细胞周期蛋白依赖性激酶4(CDK4)的蛋白表达。结果与青年组比较,模型组、对照组及实验组HDF细胞增殖能力及CDK4蛋白相对表达量均显著降低,且实验组高于模型组和对照组(P<0.05);HDF细胞β-gal活性及p16、CyclinD1蛋白相对表达量均显著升高,且实验组低于模型组和对照组(P<0.05),模型组与对照组间差异无显著性(P>0.05)。结论发酵黄芪可促进HDF细胞增殖,降低β-gal活性,同时抑制衰老相关蛋白的表达,进而发挥抗细胞衰老的作用。Objective To investigate the effects of fermented milkvetch root on proliferation and expression of aging-related proteins in human embryonic lung diploid fibroblast cell.Methods The human embryonic lung diploid fibroblasts (HDF) were cultured in vitro and divided into 4 groups according to age and dosing.Young group:HDF cells before 30 passages;model group:HDF cells after 40 passages;control group:HDF cells after 40 passages treated with 10% normal serum;test group:HDF cells after 40 passages treated with 10% fermented milkvetch root-intervention serum.Intervention began when the 40th passages was cultivated,and the morphology of HDF cells in each group was observed by optical inverted microscope.Live cell count (CCK-8) method was used to detect the proliferation of HDF cells in each group.The β-galactosidase (β-gal) activity of HDF cells in each group was detected by spectrophotometry method;protein expression of p16,Cyclin D1 (CyclinD1) and Cyclin-dependent kinase 4 (CDK4) in HDF cells was detected by Western blotting (Wb) method.Results Compared with the young group,the proliferation of HDF cells and the relative expression of CDK4 protein in the model group,the control group and the test group significantly reduced,and the test group was higher than the model group and the control group (P<0.05).The β-gal activity of HDF cells and the relative expression of p16 and CyclinD1 protein significantly increased,and the test group was lower than the model group and the control group (P<0.05),and there was no significant difference between the model group and the control group (P>0.05).Conclusion Fermented milkvetch root can promote the proliferation of HDF cells,decrease the activity of β-gal,and inhibit the expression of aging-related proteins,thereby exerting anti-aging effects.

关 键 词：人胚肺二倍体成纤维细胞 发酵黄芪 增殖 衰老 Β-半乳糖苷酶 

分 类 号：R285.5[医药卫生—中药学]