杜仲水提液对幼鼠睾丸间质细胞睾酮合成和caspase9的影响

目的:探讨杜仲水提液对幼鼠睾丸间质(Leydig)细胞睾酮合成功能和caspase9的影响。方法:原代分离、鉴定幼鼠的Leydig细胞,进行细胞培养。在加入不同剂量杜仲水提液培养24 h后,用化学发光法测定睾酮的浓度;分别在加药24 h、36 h和48 h后,用MTT法检测各组的吸光度;在加药24 h后,用流式细胞仪分析细胞凋亡率、存活率和死亡率;用实时定量反转录聚合酶链反应测定caspase9 mRNA的表达。结果:在Leydig细胞中加入杜仲水提液24 h后,上清液的睾酮浓度明显增加,与空白组比较差异有统计学意义(P<0.01);药物刺激24 h、36 h和48 h后,其吸光度与空白组比较差异均有统计学意义(P<0.01);药物刺激24 h后,细胞凋亡率和死细胞率明显降低,活细胞率明显提高,与空白组比较差异有统计学意义(P<0.01);药物刺激24 h后,caspase9 mRNA的表达量明显降低,与空白组比较差异有统计学意义(P<0.05)。结论:杜仲水提液能通过下调caspase9的表达,降低Leydig细胞的凋亡,促进其增殖,而提高其合成睾酮的能力。

Effect of Water Extraction from Eucommia Ulmoides Oliver on Testosterone Synthesis and Caspase 9 Expression of Leydig Cells in Immature Rats

Objective:To investigate the effect of water extraction from Eucommia ulmoides Oliver on testosterone synthesis and caspase 9 expression of Leydig Cells in immature Rats.Methods: Leydig cells in immature rats were cultured after primary isolation and identification.Testosterone was determined using chemiluminescence after adding different doses of Eucommia ulmoides Olive in Leydig cells 24 h.Optical density was analyzed by MTT after adding different doses of Eucommia ulmoides Oliver in Leydig cells 24 h,36 h and 48 h.Apoptosis rate was measured by flow cytometry after adding different doses of Eucommia ulmoides Olive in Leydig cells 24 h.The expression of caspase 9 mRNA was determined by Real Time-PCR.Results: The concentration of testosterone was increased significantly after Leydig cells added Eucommia ulmoides Oliver 24 h compared with that of the control group(P<0.01).The proliferation of Leydig cells sped up by adding low,medium,and high doses of Eucommia ulmoides Oliver 24 h,36 h and 48 h,of which the optical density was statistically significant compared with that of the control group(P<0.01).After Leydig cells added Eucommia ulmoides Oliver 24 h,its apoptosis rate and dead cells significantly reduced,with living cells significantly increased(P<0.01).The expression of caspase 9 mRNA was significantly decreased(P<0.05).Conclusions: Water extraction from Eucommia ulmoides Oliver can decrease the expression of caspase 9 mRNA,reduce cell apoptosis,promote proliferation and improve the synthesis of testosterone of Leydig cell in immature rats.