Effects and Mechanisms of Yangjing Capsule on Apoptosis of Mice Leydig Cell
Objective: To study the effect and potential mechanism of Yangjing Capsule for the apoptosis of Leydig cells.Method: : The mouse testis interstitial cell line MLTC-1 was used for the experiment.When the cell growth to 60% ~ 70% abundance, we add different concentrations of Yangjing capsule extract to the cells.Cell multiplication was measured by MTT assay, and the death rate and cell cycle were detected by flow cytometry.The small interference fragment (siRNA) of StAR and no-load small interfering fragments (N.C) will be transfected into MLTC-1 cell line, and after adding 1 mg / ml of Yangjing capsule we will incubate the cell for another 36 hours.The testosterone levels of supernatant were determined by radioimmunoassay; The mRNA and protein of StAR, Bax, caspase-3 and Bcl-2 will be detected after clecting the cells.When the cells grow very fast ,the cells will be transfected with plasmid DNA sequence,which containing the promoter of StAR.After adding Yangjing capsule (1 mg / ml),the cell will group for 6 hours, and we will do dual luciferase reporter gene assay to detect star gene promoter activity.When cells grow to 60% ~ 70%, we will add different concentrations of Yangjing capsule, and 36h after, cell immunofluorescence experiments will be carried out to observe intracellular localization of Star.Results: Yangjing capsule extract can significantly improve MLTC-1 cell proliferation, when the while It can reduce cell apoptosis.Yangjing capsule not only can obviously increase the contents of StAR, but also enhance the fluorescence intensity of StAR in leydig cells.What is more,Yangjing capsule can reverse the phenomenon that the content of Bcl-2 decreased, and the content of Casepase-3 and Bax increased.Conclusion: Yangjing capsule can increase the expression of StAR in MLTC-1 cells; Yangjing capsule can make the production of Bax and Caspase-3 little, and can increase the production of Bcl-2 by upregulating the expression of StAR.
Yangjing capsule StAR Bax Casepase-3 Bcl-2 Leydig cells
Chen Guanghui Jin Baofang Sun Dalin Cai Bin
Andrology Department of Integrative Medicine, Zhongda Hospital, School of Medicine, Southeast University, Nanjing 210009, China
国内会议
济南
英文
868-869
2016-09-01（万方平台首次上网日期，不代表论文的发表时间）