摘要
目的探讨养精胶囊改善生精功能进而治疗男性不育症可能的作用机制。方法在GC-1细胞中加入养精胶囊提取液,分为空白组和低、中、高剂量组,在作用24、48、72 h后采用MTT法检测各组的吸光值;作用48 h后,采用流式细胞仪检测细胞周期及细胞凋亡;用实时荧光定量PCR法和Western blot法检测凋亡相关基因caspase-3的表达。结果在作用24、48、72 h后,养精胶囊提取液低、中、高剂量组细胞增殖明显加快,其吸光度与同期空白组相比有统计学意义(P<0.01);养精胶囊提取液低、中、高剂量组作用GC-1细胞48 h后,S期细胞比例逐渐增加,与空白组相比有统计学意义(P<0.05);细胞凋亡率明显降低,与空白组相比有统计学意义(P<0.01);caspase-3mRNA表达呈下降趋势,中、高剂量组与空白组相比有统计学意义(P<0.01);caspase-3蛋白表达呈下降趋势,高剂量组与空白组相比有统计学意义(P<0.01)。结论养精胶囊具有促进GC-1细胞增殖、抑制细胞的凋亡,降低caspase-3的表达水平的作用,且呈剂量依赖性。
Objective To discuss the possible mechanism of Yangjing Capsules in improving spermatogenic function and then treating male infertility. Methods The extracting solution of Yangjing Capsules in different doses were added into GC-1 cells and the cells were divided into blank group, low- dose group, mid-dose group and high-dose group. The absorbanee values in all groups were detected by using methyl thiazolyl tetrazolium test （MTr） respectively after 24 h, 48 h and 72 h. The cell cycle and apoptosis were examined by using flow cytometer after 48 h, and the expression of caspase-3 （ apoptosis- related gene） was detected by using real-time fluorescence quantitative polymerase chain reaction （RT- PCR） and Western blot method. Results After 24 h, 48 h and 72 h, the proliferation of GC-1 cells was speeded up significantly in low-dose group, mid-dose group and high-dose group, and the comparison in absorbance had a statistical significance between blank group and medicated groups （ P 〈 0.01 ）. After 48 h, the proportion of S phase cells increased gradually （ P 〈 0.05 ） and apoptosis rate decreased significantly （P 〈 0.01 ） in low-dose group, mid-dose group and high-dose group and had statistical significance compared with blank group. The expression of caspase-3 mRNA showed a decreasing tendency with a statistical significance compared between mid-dose group or high-dose group and blank group （P 〈0.01 ）, and expression of caspase-3 protein showed a decreasing tendency with a statistical significance compared between high-dose group and blank group （ P 〈 0.01 ）. Conclusion Yangjing Capsules has the effects of improving the proliferation of GC-1 cells＇ inhibiting apoptosis and reducing caspase-3 expression in a dose-dependent manner.
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2014年第7期500-504,I0004,共6页
Journal of Beijing University of Traditional Chinese Medicine
基金
国家自然科学基金项目(No.81273760/H2709)
江苏省研究生培养创新项目(No.CXLX13_583)
