知母皂苷元对性早熟大鼠下丘脑Kiss-1和GPR54 mRNA表达的影响

Effects of anemarrhenae saponin on the expression of Kiss-1 and GPR54 mRNA in hypothalamus of rats with precocious puberty

目的探讨知母皂苷元对性早熟大鼠的治疗作用及可能的作用机制。方法将24只SD大鼠分为正常组、模型组、亮丙瑞林组和知母皂苷元组,每组6只。5日龄大鼠皮下注射达那唑300μg,建立性早熟模型,造模10d后进行药物干预。采用苏木精-伊红(HE)染色观察子宫和卵巢发育情况;酶联免疫吸附试验(ELISA)法测定血清黄体生成素(LH)、卵泡刺激素(FSH)和雌二醇(E_(2))水平;实时荧光定量聚合酶链反应(RT-qPCR)法检测促性腺激素释放激素(GnRH)、亲吻素(Kiss-1)和G蛋白偶联受体54(GPR54)mRNA表达水平。结果与模型组比较,知母皂苷元组的阴道开口时间更长,子宫和卵巢系数更小,子宫壁厚度更薄,差异具有统计学意义(P<0.05);与模型组比较,亮丙瑞林组和知母皂苷元组LH、FSH、E_(2)水平更低,差异具有统计学意义(P<0.05);与模型组比较,亮丙瑞林组和知母皂苷元组Kiss-1 mRNA表达水平更低,差异具有统计学意义(P<0.05);与模型组比较,知母皂苷元组GPR54mRNA表达水平更低,差异具有统计学意义(P<0.05)。结论知母皂苷元可能通过Kiss-1与GPR54信号通路调节下丘脑-垂体-性腺轴(HPG轴)来治疗性早熟大鼠。

Objective To investigate the therapeutic effect of anemarrhena saponin on precocious puberty rats and its possible mechanism.Methods 24 SD rats were divided into normal group, model group, leuprorelin group and anemarrhena saponin group, with 6 rats in each group. The model of precocious puberty was established by subcutaneous injection of danazol 300μg into 5-day-old rats, and drug intervention was carried out 10 days later. The development of uterus and ovary was observed by hematoxylin eosin(HE)staining. The serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH) and estradiol (E_(2)) were determined by enzyme linked immunosorbent assay(ELISA). The mRNA expression levels of gonadotropin-releasing hormone (GnRH), Kiss-1 and G protein-coupled receptor 54(GPR54) were detected by real time fluorescence quantitative polymerase chain reaction ( RT-qPCR).Results Compared with the model group, the vaginal opening time of anemarrhena saponin group was longer, the coefficient of uterus and ovary was smaller, and the thickness of uterine wall was thinner, the difference was statistically significant (P< 0. 05). Compared with the model group, the levels of LH, FSH and E_(2) in leuprorelin group and anemarrhena saponin group were lower,and the difference was statistically significant (P< 0. 05). Compared with the model group, the expression level of Kiss-1 mRNA in leuprorelin group and anemarrhena saponin group was lower, and the difference was statistically significant (P< 0. 05). Compared with the model group, the GPR54 mRNA expression level of anemarrhena saponin group was lower, and the difference was statistically significant (P< 0. 05).Conclusions Anemarrhena saponin may regulate hypothalamic pituitary gonadal axis (HPG axis) through Kiss-1 / GPR54 signaling pathway in the treatment of precocious puberty.