急性疼痛应激状态下SD大鼠睾丸内膜联蛋白A5的表达变化

目的 观察急性疼痛应激状态下,睾丸中annexin 5表达变化.方法 福尔马林注射建立急性疼痛模型,分别在注射后1 h,6 h,24 h,72 h取睾丸,免疫组织化学和western b10t分析annexin 5免疫定位和蛋向表达,荧光定量PCR方法检测annexin 5 mRNA水平变化.结果 免疫组化显示,annexin 5在急性应激组和对照组中定位在间质细胞、支持细胞,应激组与对照组相比定位无明显的变化.Westem b1ot分析发现疼痛应激1 h和6 h后,annexin 5的表达分别降低16.9%和12.8%,而在疼痛应激24 h和72 h后,annexin 5的表达分别升高33.7%和25%.组间比较发现,在l h和24 h时annexjn 5表达较对照组差异有统计学意义(P<0.05).荧光定量方法检测annexin 5 mRNA水平变化,发现疼痛应激1h和6 h annexin 5 mRNA相对含量降低,随后在24 h和72 h升高,但各组间差异均无统计学意义.结论 急性疼痛应激主要影响annexin 5蛋白的表达,而不影响annexin 5的转录.Annexin 5作为睾丸应激因子之一可能介导了应激对生殖功能的抑制作用.

Study on the expression of annexin 5 in rat testis under acute pain stress

Objective To study on the expression of annexin 5 expression in rat testis under acute pain stress. Methods SD rat models with acute pain stress were established by injection of formalin, and their testis were collected respectively at 1 h ,6 h ,24 h and 72 h after injection. The distribution and expression of annexin 3 protein were detected by Immunohistochemistry and western blot. The level of annexin 5 mRNA was measured by quantitative RT-PCR. Results Immunohistochemistry staining showed that annexin 5 was mainly distributed in Sertoli cell and Leydig cells of rat testis. Analysis of western blot exhibited that the expression of annexin 5 decreased about 16.9 % and 12. 8% respectively at lh and 6h after injection of formalin, whereas increased 33.7% and 25% at 24 h and 72 h after injecting formalin. Compared with that of control group, statistical difference were found in the expression of annexin 5 at lh and 24 h. The mRNA level of annexin 5 had a similar expression pattern with its protein expression at different time points. However, no distinct difference in mRNA level of annexin 5 was found among groups. Conclusion Acute pain stress might mainly affect the protein level of annexin 5 in testis, but no mRNA level of annexin 5, which indicated that annexin S might mediated the process of stress inhibiting reproduction, as a stress response gene.