目的 通过氯米芬联合补肾种子汤调控"同源框基因A10(HOXA-10)"表达水平,研究其影响多囊卵巢综合征(Polycysticovarysyndrome,PCOS)雌鼠子宫内膜容受性的相关机制.方法 将40只SPF级SD雌性大鼠按随机数字表法分为空白对照组、模型组、氯米芬组、补肾种子汤组、氯米芬联合补肾种子汤组,每组各8只.通过丙酸睾酮+绒毛膜促性腺激素(Human chorionic gonadotropin,HCG)颈背部皮下注射SD雌鼠构建PCOS动物模型.造模成功后予以氯米芬、补肾种子汤、氯米芬联合补肾种子汤药物干预,连续14 d,期间行阴道脱落涂片,并绘制大鼠体质量增长变化曲线.大鼠处死后,Elisa检测血清性激素雌二醇(Estradiol,E2)、孕酮(Progesterone,P)、促黄体素(Luteinizinghormone,LH)、促卵泡素(Follical stimulating hormone,FSH)含量,行卵巢及子宫 HE 染色,免疫组化观察HOXA-10在子宫组织中表达水平.结果 氯米芬组、氯米芬联合补肾种子汤组与模型组比较,性激素E2、P值升高,LH/FSH比值降低,差异有统计学意义(P<0.01);氯米芬联合补肾种子汤组与氯米芬组比较,性激素E2、P值升高,LH/FSH比值降低,差异有统计学意义(P<0.05),与补肾种子汤组比较,性激素E2值升高,差异有统计学意义(P<0.01),性激素P值升高,LH/FSH比值降低,差异有统计学意义(P<0.05).卵巢HE染色见:卵巢囊状卵泡明显减少,卵泡形态稍欠规则,颗粒细胞层增厚,见少量黄体.子宫HE染色见:子宫内膜略显平坦,内膜增厚,子宫腺体增多,管腔轻度扩张,分泌物增多.氯米芬联合补肾种子汤组雌鼠HOXA-10表达水平明显高于模型组、氯米芬组及补肾种子汤组,差异有统计学意义(P<0.05).以上指标数值增减及子宫、卵巢形态学改变以氯米芬联合补肾种子汤组最为明显,疗效最为显著.结论 氯米芬联合补肾种子汤治疗PCOS雌鼠模型,疗效明显优于单纯使用氯米芬或补肾种子汤,其作用机制为通过下调LH/FSH比值,上调血清E2、P水平及子宫组织HOXA-10表达水平,改善PCOS雌鼠卵巢形态学,提升大鼠子宫内膜容受性.
Objective To regulate the expression level of"homeobox gene A10(HOXA-10)"by an improved method and study the mechanism of the improved method on endometrial receptivity of female rats with polycysticovarysyn-drome(PCOS).Method A total of 40 SD female rats at SPF level were randomly divided into a control group,PCOS model group,PCOS model+clomiphene group,PCOS model+kidney tonifying seed soup group,PCOS model+improve-ment group,with eight rats in each group.The PCOS animal model was established by neck and back subcutaneous injec-tion of testosterone propionate and human chorionic gonadotropin(HCG).After successful mold making,the rats were inter-vened with clomiphene,kidney tonifying seed soup,and a modified method(clomiphene+kidney tonifying seed soup)for 14 days,during which vaginal shedding smear was taken,and the curve of weight growth of rats was drawn.After rats were kill-ed,Elisa tested the content of serum sex hormone estradiol(E2),progesterone(P),luteinizinghormone(LH),and follicular stimulating hormone(FSH).HE staining of the ovary and uterus was performed,and the expression level of HOXA-10 in uterine tissue was observed by immunohistochemistry.Results Compared with the PCOS model+clomiphene group,PCOS model+improvement group,and PC OS model group,E2 and P values increased,and LH/FSH ratio decreased.The differ-ence was statistically significant(P<0.01).Compared with the PCOS model+clomiphene group,E2 and P values of the P-COS model+improvement group increased,and LH/FSH ratio decreased.The difference was statistically significant(P<0.05).Compared with the PCOS model+kidney tonifying seed soup group,the value of E2 was increased(P<0.01),and that of P was increased.The ratio of LH/FSH was decreased(P<0.05).HE staining of the ovary showed that the cystic follicles of the ovary were significantly reduced,and the shape of the follicles was slightly irregular.The granulosa cell layer was thickened,and a small amount of corpus luteum was seen.HE staining of the uterus showed that the endometrium was slightly flat and thickened,and the uterine glands increased.The lumen was slightly dilated,and the secretion increased.The expression level of HOXA-10 in the PCOS model+improvement group was significantly higher than the PCOS model group,PCOS model+clomiphene group,and PCOS model+kidney tonifying seed soup group(P<0.05).The changes of the above indexes and the morphological changes of the uterus and ovary were most obvious in the PCOS model+improve-ment group.This meant that the PCOS model+improvement group had the best effect.Conclusion The improved method is superior to clomiphene or kidney tonifying seed soup alone in the treatment of PCOS female rats.The mechanism of the improved method is to improve the ovarian morphology and increase the endometrial receptivity of PCOS female rats by downregulating the LH/FSH ratio and upregulating the serum expression levels of E2,P,and HOXA-10 in uterus tissue.