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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,6h,24 h,72 h取睾丸,免疫组织化学和western blot分析annexin 5免疫定位和蛋白表达,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。Western blot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1 h和24 h时annexin 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1 h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。
Objective To observe the expression of annexin 5 in testis under acute pain stress. Methods Acute pain models were established by formalin injection. Testes were harvested at 1 h, 6 h, 24 h and 72 h after injection respectively. Immunohistochemistry and western blot were used to analyze the immunolocalization and protein expression of annexin 5. The levels of annexin 5 mRNA level changes. Results Immunohistochemistry showed that annexin 5 localized in stromal cells and supporting cells in acute stress group and control group. There was no significant difference in the localization of annexin 5 between the stress group and the control group. Western blot analysis showed that the expression of annexin 5 decreased by 16.9% and 12.8% after 1 h and 6 h of pain stress, respectively, while the expression of annexin 5 increased by 33.7% and 25% after 24 h and 72 h of pain stress respectively. . Compared between two groups, annexin 5 expression at 1 h and 24 h was significantly different from that in control group (P <0.05). The changes of annexin 5 mRNA level were detected by fluorescence quantitative assay. The relative contents of annexin 5 mRNA decreased in 1 h and 6 h after pain stress, and then increased at 24 h and 72 h. However, there was no significant difference among the groups. Conclusions Acute pain stress mainly affects the expression of annexin 5 without affecting the transcription of annexin 5. Annexin 5 as a testicular stress factor may mediate the inhibitory effect of stress on reproductive function.