蛤蚧乙醇提取液对大鼠卵巢颗粒细胞凋亡的影响

The Effect of the Gekko Gecko Ethanol Extract on the Granular Cells Apoptosis of Rat Ovaries

目的:研究蛤蚧乙醇提取液对大鼠卵巢颗粒细胞凋亡的影响。方法:3月龄和6月龄Wistar雌性大鼠各20只,每个月龄组随机分为实验组和对照组,每组各10只。实验组以蛤蚧乙醇提取液2g(原药)/kg.d灌胃,对照组以体积相当的0.9%氯化钠液灌胃,15日后取各组大鼠卵巢经脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)和碘化丙啶(PI)染色、磷脂结合蛋白V/碘化丙啶(AnnexinV/PI)双染经流式细胞术(FCM)检测颗粒细胞凋亡及其周期分布情况。结果:TUNEL检测颗粒细胞凋亡数3月龄和6月龄实验组分别是34±7个和53±11个,对照组分别为48±9个和76±17个,差异有高度统计学意义(P<0.01);PI检测的颗粒细胞凋亡率3月龄和6月龄实验组分别为(6.06±2.01)%和(12.16±3.26)%,对照组分别为(8.23±2.13)%和(23.69±6.28)%,差异有统计学意义(P<0.05),但是细胞周期的分布差异却无统计学意义(P>0.05);AnnexinV/PI检测的颗粒细胞早期凋亡率3月龄和6月龄实验组分别为(6.71±3.11)%和(23.74±6.28)%,对照组分别为(19.05±5.78)%和(36.55±8.35)%,差异有高度统计学意义(P<0.01)。3月龄和6月龄实验组大鼠卵巢颗粒细胞凋亡均明显少于对照组(P<0.01或P<0.05)。结论:蛤蚧乙醇提取液能有效抑制大鼠卵巢颗粒细胞的凋亡,从而改善大鼠卵巢功能,并可能由此延缓大鼠卵巢的衰老。

Objective：To study the effects of the Gekko gecko ethanol extract on granular cells apoptosis of rat ovaries.Methods：3 month and 6 month female Wistar rats were taken in this study,and 20 rats were in each age group.Each age group were randomly divided into two groups（experimental group and control group）10 rats for each group.2g（active compound）kg·d of Gekko gecko ethanol extract was given by intragastric administration in experimental group,corresponding volume of 0.9% sodium chloride was given in control group by the same way.Apoptosis of granular cells and distribution of cell cycle in rat ovary were measured by terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling（TUNEL）and propidium iodide（PI）staining,AnnexinVPI double labelling staining by flow cytometry（FCM）after administration for 15 days.Results：The apoptosis numbers of granular cells in 3 month and 6 month age experimental group were 34±7 and 53±11,in control group were 48±9 and 76±17 respectively by TUNEL.There was significantly statistic difference（P0.01）.The apoptosis rates in 3 month and 6 month age experimental group were（6.06±2.01）% and（12.16±3.26）%,in the control group were（8.23±2.13）% and（23.69±6.28）% respectively by PI staining.There was significantly statistic difference（P0.05）.However,no significant difference was found between two groups in cellular cycle distribution（P0.05）.Early apoptosis rates in 3 month and 6 month age experimental group were（6.71±3.11）% and（23.74±6.28）%,in the control group were（19.05±5.78）% and（36.55±8.35）% respectively by AnnexinVPI double labelling staining FCM.There was significantly statistic difference（P0.01）.The apoptosis of granular cells were significantly less in experimental groups comparing with control groups both in 3 month and 6 month age group（P0.01 or P0.05）.Conclusions：The ethanol extract of gekko gecko can inhibit the apoptosis of granular cells in rat ovary,then improves the ovary function and delay ageing of rat.