摘要
目的探讨补肾活血汤对衰老耳蜗毛细胞株HEI-OC1自噬和凋亡的影响。方法应用D-半乳糖构建衰老HEI-OC1细胞的体外模型后,将细胞分为对照组、衰老组、含药血清低、中及高剂量组(5%、10%、20%),采用噻唑蓝(MTT)比色法检测耳蜗HEI-OC1细胞的增殖能力;β-半乳糖苷酶染色检测细胞衰老状态;膜联蛋白V/碘化丙啶(Annexin V-FITC/PI)双染流式细胞术检测细胞凋亡的程度;Western印迹检测细胞中自噬和凋亡相关蛋白的表达水平。结果MTT、β-半乳糖苷酶染色、Annexin V-FITC/PI双染流式细胞术和Western印迹结果显示,与对照组比较,衰老组HEI-OC1细胞增殖能力明显下降;细胞衰老状态明显增加,细胞凋亡率明显增加,Atg5、Atg12、Beclin1和Bcl-2蛋白表达水平明显下降,p62和Bax蛋白表达水平显著升高(均P<0.01)。与衰老组比较,补肾活血汤各组HEI-OC1细胞增殖能力明显上升;细胞凋亡率明显降低;Atg5、Atg12、Beclin1和Bcl-2蛋白表达水平显著增加(均P<0.05);p62和Bax蛋白表达水平显著下降(均P<0.05),且均呈现出剂量依赖性。结论补肾活血汤可通过促进自噬及抑制凋亡来延缓耳蜗毛细胞株HEI-OC1的衰老,且与其浓度相关。
Objective To explore the effect of Bushen Huoxue Decoction(BSHXT)on autophagy and apoptosis of aging cochlear hair cell line HEI-OC1.Methods After the aging model of HEI-OC1 cells was established by D-galactose in vitro,the cells were divided into control group,aging group,low,medium and high dose groups(5%,10%and 20%)of serum containing BSHXT Huoxue Decoction.MTT assay was used to detect the proliferation ability of HEI-OC1 cells in cochlea.β-galactosidase staining was used to detect the aging state of cells.Apoptosis was detected by Annexin V-FITC/PI double staining flow cytometry.Western blot analysis was used to detect the expression level of autophagy and apoptosis-related proteins in cells.Results The results of MTT,β-galactosidase staining,Annexin V-FITC/PI double staining flow cytometry and Western blot showed that compared with the control group,the proliferation ability of HEI-OC1 cells in aging group was significantly decreased,and the aging state of cells was significantly increased(P<0.01).The apoptosis rate was significantly increased(P<0.01),the expression levels of Atg5,Atg12,Beclin1 and Bcl-2 were significantly decreased(P<0.01),and the expression levels of p62 and Bax were significantly increased(P<0.01).Compared with aging group,the proliferation ability of HEI-OC1 cells in Bushen Huoxue Decoction groups was significantly increased(P<0.05),the apoptosis rate was significantly decreased(P<0.05),the expression levels of Atg5,Atg12,Beclin1 and Bcl-2 protein were significantly increased(P<0.05),the expression levels of p62 and Bax protein were significantly decreased(P<0.05),and both showed dose dependence.Conclusions BHXT Huoxue Decoction could delay the aging of cochlear hair cell line HEI-OC1 by promoting autophagy and inhibiting apoptosis,which is related to its concentration.
作者
郭向东
王青玲
张梦娴
王庆林
GUO Xiang-Dong;WANG Qing-Ling;ZHANG Meng-Xian(Department of Otolaryngology,The First Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450000,Henan,China)
出处
《中国老年学杂志》
CAS
北大核心
2022年第2期371-376,共6页
Chinese Journal of Gerontology
基金
国家自然科学基金(81403439)
河南省科技攻关计划(192102310170,202102310174)
河南省中医药科学研究专项重点课题(20-21ZY1045)。
