GnRHa通过影响大鼠子宫内膜异位症Tspan5的表达影响内膜容受性

GnRHa affects the expression of Tspan5 in rat endometriosis affect endometrial receptivity

目的 探讨Tspan5和OPN与子宫内膜容受性的相关性,阐明Tspan5作为评估内异症患者子宫内膜容受性的参考指标的意义。方法 40只实验大鼠随机分为GnRHa降调模型组(n=20)和对照组(n=20)。GnRHa降调模型组:按0.04 mg/kg进行肌内注射,注射浓度为0.25 mg/mL的长效GnRHa的生理盐水1 d,28 d在当日17点按1∶1的比例和正常雄鼠合笼,第2天早晨8点检查,出现阴栓标为怀孕第1日,按此类推。对照组以等量生理盐水代替GnRHa,余同GnRHa降调模型组。子宫内膜标本进行WB、real time-PCR和免疫组化以检测Tspan5和OPN的mRNA表达和蛋白定位。ELISA法检测血清E2、P、CA125水平,分析Tspan5和OPN与血清E2、P、CA125水平的相关性。结果 Q-PCR、免疫组化和免疫印迹均显示,GnRHa降调模型组GnRHa降调前子宫内膜中Tspan5及OPN蛋白的表达显著高于对照组,差异有统计学意义(P<0.05)。GnRHa降调模型组GnRHa降调后Tspan5及OPN的表达明显下降,接近对照组的表达量(P>0.05)。ELISA法检测血清中E2、P和CA125的表达水平,显示对照组子宫内膜中E2、P和CA125的表达显著高于GnRHa降调模型组(P<0.05)。结论 Tspan5在内异症大鼠的子宫内膜显著高表达,其可能是子宫内膜异位症影响内膜容受性的因素。

Objective To investigate the correlation between Tspan5 and OPN and endometrial receptivity, and to clarify the significance of Tspan5 as a reference index for evaluating endometrial receptivity in patients with endometriosis.Methods Forty experimental rats were randomly divided into GnRHa tone lowering model group(n=20) and control group(n=20). GnRHa lowering model group: Intramuscular injection of 0.04 mg/kg, injection of 0.25 mg/mL long-acting GnRHa normal saline for 1 day, cage with normal male rats in the ratio of 1∶1 at 17:00 on the same day on the 28th day, check the vaginal suppository at 8:00 on the next morning, and see that the vaginal suppository is recorded as the first day of pregnancy, and so on. Control group: The same amount of normal saline was used to replace GnRHa, the rest is the same as the GnRHa down regulation model group. Endometrial specimens were subjected to WB, real time PCR and immunohistochemistry to detect the mRNA expression and protein localization of tspan5 and OPN. The levels of serum E2, P and CA125 were detected by ELISA,and the correlation between tspan5 and OPN and the levels of serum E2, P and CA125 was analyzed. Results Q-PCR, immunohistochemistry, and immunoblotting showed that the expression of Tspan5 and OPN proteins in the endometrium of the GnRHa lowering model group before GnRHa down-regulation was significantly higher than that of the control group, and the difference was statistically significant(P<0.05). The expression of Tspan5 and OPN in the GnRHa lowering model group decreased significantly after the down-regulation of GnRHa, which was close to the expression level of the control group(P>0.05). The expression levels of E2, P and CA125 in serum were detected by ELISA, which showed that the expression levels of E2, P and CA125 in endometrium in control group were higher than those in GnRHa down-regulated model group(P<0.05). Conclusion Tspan5 was significantly overexpressed in endometrium of endometriosis rats, which may be a factor affecting endometriosis receptivity.