电针对促排卵大鼠子宫VEGF、VEGFR2表达的影响
The Effect of Electroacupuncture on the Expression of VEGF and VEGFR2 in the Uterus of the Rat Model of Ovulation Induction
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摘要: 目的 观察电针刺激对激素促排卵大鼠子宫组织中VEGF、VEGFR2表达的影响,并探讨其作用机制。方法 10周龄SD雌性大鼠30只,随机分为对照组、模型组及治疗组,每组各10只。模型组和治疗组腹腔注射孕马血清促性腺激素(PMSG)20U,48h后注射同等剂量的人绒毛膜促性腺激素(hCG),与种鼠合笼,次晨分笼造模,治疗组在与种鼠合笼前1周连续电针治疗7d(刺激强度1mA,疏密波2/15Hz,持续刺激15min)。合笼后第3.5天处死实验动物,留取子宫组织,观察形态改变;采用免疫组化方法检测VEGF、VEGFR2蛋白的表达;采用qPCR技术测定子宫组织中VEGF、VEGFR2mRNA的表达,采用2 -△△Ct 方法计算。结果 肉眼观察对照组子宫形态规整,表面红润光滑;模型组子宫形态肿大扭曲,表面色泽暗红充血;治疗组子宫形态略微肿胀,表面红润,无充血现象。免疫组化可见VEGF、VEGFR2蛋白在3组子宫内膜上均有表达,平均光密度没有明显差异,但表达分布部位有差异。模型组相比于对照组,在基质细胞和腺体上的表达分布较为广泛。VEGFR2在子宫的肌层组织中表达有明显差异:对照组几乎没有表达,模型组表达则明显增高(P<0.05),治疗组比模型组表达有明显下降(P<0.05)。模型组子宫内膜组织VEGFmRNA的表达是对照组的0.68倍,下降趋势明显;治疗组是对照组的0.88倍,有上升趋势。模型组子宫内膜组织VEGFR2mRNA的表达是对照组的0.46倍,下降趋势明显;治疗组是对照组的0.82倍,有比较明显的上升趋势。结论 电针可以调整促排卵大鼠子宫组织中VEGF及其受体的表达,从而调整激素促排卵导致的子宫内膜种植窗口期的同步性。Abstract: OBJECTIVE To observe the effect of electroacupuncture on the expression of VEGF and VEGFR2 in the uterus of the rat model of ovulation induction and to investigate the possible mechanism. METHODS Female SD rats aged 10 weeks were divided into control, model and treat groups randomly, 10 in each group. The model group and the treatment group were peritoneal injected with pregnant horse serum gonadotrophin (PMSG) of 20U and injected of the same dose of human chorionic gonadotropin(hCG) after 48h, and then cohabitated with male SD nice until next morning. A week before cohabitation, the treat group received a consecutive electroacupuncture treatment for 7d(dilatational wave of 2/15Hz for 30min and stimulative intensity of 1mA). The animals were sacrificed 3.5d after the cohabitation. The uterine tissue was taken to observe the morphological change, immunehistochemical method was used to inspect the expression of VEGF and VEGFR2, and qPCR technology was used to determine the expression of VEGF and VEGFR, with the calculation by 2 -△△Ct . RESULTS Macroscopic observation: the surface in control group was regular and smooth; the model group was intumescentia, dull-red and hyperaemia; the treatment group was slightly swollen, rosy and no congestion. The expression of VEGF and VEGFR2 were observed in the 3 groups by immunohistochemical method, with no significant difference in average optical density, but differences in the expression distribution. Compared with the control group, the expression distribution of stromal cells and glands in the model group was more extensive, and the expression of VEGFR2 in myometrium increased significantly(P<0.05). The electroacupuncture treatment inhibited the the expression of VEGFR2 significantly(P<0.05). The expression of VEGF mRNA in uterine tissues of model group was 0.68 times of control group, with significant declining trend; the treat group was 0.88 times of model group, with significant increasing trend. The expression of VEGF2 mRNA in uterine tissues of model group was 0.46 times of control group, with significant declining trend; the treat group was 0.82 times of model group, with significant increasing trend. CONCLUSION The electroacupuncture can adjust the expression of VEGF and its receptor in the uterine tissue of ovulation induction rat model, thus regulate the synchronization of the endometrial planting window caused by hormone.