论文部分内容阅读
目的:探讨Chk1基因在人精子中的表达及其临床意义。方法:收集正常组、少精子组、弱精子组和少弱精子组的标本各20份,分别采用Western印迹和RT-PCR方法检测各组精子中Chk1的表达,并计算其相对表达量;采用DNA梯度法检测4组精子中DNA的损伤和梯度变化,用Annexin V/PI双染色法检测各组精子的凋亡情况。结果:Chk1基因在正常组和其他3组的精子中均有表达,Chk1在各组中表达有差异,在正常组、少精子组、弱精子组和少弱精子组的Chk1 mRNA的相对表达量分别为1.00±0.22、0.76±0.10、0.45±0.08、0.37±0.07,各组间比较差异有显著性(P<0.01)。CHK1蛋白的相对表达量与mRNA类似。DNA梯度检测显示弱精子组和少弱精子组中DNA呈现梯状条带;Annexin V/PI双染色法检测发现,与正常组相比,少精子组、弱精子组、少弱精子组和100%d级精子组中精子凋亡率明显增加[分别为(7.6±0.34)%、(8.3±0.60)%、(11.6±0.92)%、(12.5±1.43)%和(17.0±1.98)%,各组间比较P<0.05]。结论:实验证实Chk1在人类精子中有表达,且在不同患者中的表达显著不同,Chk1的表达与DNA损伤和精子凋亡之间存在相关性。即Chk1表达的减少可能导致精子DNA损伤修复功能的下降,精子凋亡增加,从而影响精液质量。
Objective: To investigate the expression of Chk1 gene in human sperm and its clinical significance. Methods: 20 samples of normal group, oligospermia group, weak sperm group and oligoasthenospermia group were collected. The expression of Chk1 was detected by Western blot and RT-PCR respectively, and the relative expression of Chk1 was calculated. DNA gradient was used to detect DNA damage and gradient in four groups of spermatozoa. Apoptosis of each group was detected by Annexin V / PI double staining. Results: Chk1 gene was expressed in normal and other three groups of spermatozoa, Chk1 expression was different in each group, the relative expression of Chk1 mRNA in normal group, oligospermia group, weak sperm group and less weak sperm group Respectively, 1.00 ± 0.22,0.76 ± 0.10,0.45 ± 0.08,0.37 ± 0.07, the difference between the groups was significant (P <0.01). The relative expression level of CHK1 protein is similar to that of mRNA. The results of DNA ladder showed that the DNA in weak sperm group and weak sperm group showed ladder-like bands. The results of Annexin V / PI double staining showed that in oligospermia group, weak sperm group, oligo-weak sperm group and 100 The apoptosis rate of sperm in% d sperm group was significantly increased (7.6 ± 0.34%, (8.3 ± 0.60)%, (11.6 ± 0.92)%, (12.5 ± 1.43)% and (17.0 ± 1.98)%, Comparison between groups P <0.05]. Conclusion: Chk1 was expressed in human spermatozoa and significantly different in different patients. Chk1 expression was correlated with DNA damage and sperm apoptosis. That reduction of Chk1 expression may result in the decline of sperm DNA damage repair function, increased sperm apoptosis, thus affecting the semen quality.