知母总皂苷对雌性性早熟SD大鼠性激素水平的影响及对HPGA轴的作用研究

Effects of anemarrhena total saponins on sex hormone levels in female SD rats with precocious puberty and its effects on HPGA axis

关键词:  知母总皂苷  雌性  性早熟  性激素  下丘脑-垂体-性腺轴

Key words:  anemarrhena total saponins  female  precocious puberty  sex hormone  hypothalamic-pituitary-gonadal axis

摘要:

[目的] 探讨知母总皂苷对雌性性早熟SD大鼠性激素水平的影响及对下丘脑-垂体-性腺轴（HPGA）的作用。[方法] 建造雌性性早熟SD大鼠模型，随机分为模型组、知母总皂苷低剂量组（知低组）、知母总皂苷高剂量组（知高组）、阳性对照组，另设正常组。经相应干预后，记录各组大鼠阴门开启时间与首个动情间期时间；取大鼠性器官，计算子宫、卵巢指数；HE染色观察大鼠卵巢形态学变化；ELISA检测大鼠血清性激素卵泡生成激素（FSH）、黄体生成激素（LH）、雌二醇（E2）水平；qPCR和Western blot检测大鼠下丘脑肿瘤转移抑制基因亲吻素-1（KISS-1）、G蛋白偶联受体54（GPR54）、促性腺激素释放激素（GnRH）及其受体（GnRHR） mRNA与蛋白表达。[结果] 与正常组比较，模型组大鼠阴门开启时间与首个动情间期时间较短，子宫、卵巢指数较高，黄体数量较多，血清FSH、LH、E2水平较高，下丘脑KISS-1、GPR54、GnRH、GnRHR mRNA与蛋白表达量较高（P<0.05）；与模型组比较，知低组、知高组、阳性对照组大鼠阴门开启时间与首个动情间期时间较长，子宫、卵巢指数较低，黄体数量较少，血清FSH、LH、E2水平较低，下丘脑KISS-1、GPR54、GnRH、GnRHR mRNA与蛋白表达量较低（P<0.05）；与知低组比较，知高组、阳性对照组大鼠阴门开启时间与首个动情间期时间较长，子宫、卵巢指数较低，黄体数量较少，血清FSH、LH、E2水平较低，下丘脑KISS-1、GPR54、GnRH、GnRHR mRNA与蛋白表达量较低（P<0.05）；与知高组比较，阳性对照组大鼠阴门开启时间与首个动情间期时间较长，子宫、卵巢指数较低，黄体数量较少，血清FSH、LH、E2水平较低，下丘脑KISS-1、GPR54、GnRH、GnRHR mRNA与蛋白表达量较低（P<0.05）。[结论] 知母总皂苷可调节雌性性早熟SD大鼠性激素水平，对HPGA轴有抑制作用。

Abstract:

[Objective] To observe the effect of anemarrhena total saponins on the level of sex hormones in female SD rats with precocious puberty and the effect on the hypothalamus-pituitary-gonad axis （HPGA）. [Methods] A female precocious SD rat model was constructed and randomly divided into a model group，an anemarrhena total saponins low-dose group （Zhi-low group），an anemarrhena total saponins high-dose group （Zhi-high group），a positive control group，and a normal group. After corresponding intervention，record the opening time of the vulva and the time of the first estrus in each group of rats. Take the sex organs of the rats to calculate the uterus and ovarian indexes. HE staining to observe the morphological changes of the rat ovary. ELISA to detect serum levels of sex hormones follicle-forming hormone （FSH），luteinizing hormone （LH） and estradiol （E2） in rats. QPCR and Western blot detection of rat hypothalamic tumor metastasis suppressor genes kissin-1 （KISS-1），G protein-coupled receptor 54 （GPR54），gonadotropin releasing hormone （GnRH） and its receptor （GnRHR） mRNA and protein expression. [Results] Compared with the normal group，the vulva opening time and the first estrus interval time of the model group were shorter，the uterus and ovary indexes were higher，the number of corpus luteum was higher，the serum FSH，LH，and E2 levels were higher，and the upper thalamus KISS-1，GPR54，GnRH，GnRHR mRNA and protein expression levels were higher （P<0.05），compared with the model group，the vulva opening time and the first estrus interval time were longer in the Zhi-low group，Zhi-high group，and positive control group. Uterus and ovary index is low，the number of corpus luteum is low，serum FSH，LH，E2 levels are low，and the expression of KISS-1，GPR54，GnRH，GnRHR mRNA and protein in the epithalamus is low （P<0.05），in comparison，the opening time of the vulva in the Zhi-high group and the positive control group was longer than the first estrus interval，the uterus and ovarian indexes were lower，the number of corpus luteum was lower，the serum FSH，LH，and E2 levels were lower，and the hypothalamus KISS-1，GPR54，GnRH，GnRHR mRNA and protein expression are lower （P<0.05），compared with Zhi-high group，the time of opening the vulva and the first estrus interval is longer in the positive control group，and the index of uterus and ovary is lower，the number of corpus luteum is small，the level of serum FSH，LH，E2 is low，and the expression of KISS-1，GPR54，GnRH，GnRHR mRNA and protein in the upper thalamus is low （P<0.05）. [Conclusion] The anemarrhena total saponins can regulate the level of sex hormones in female precocious SD rats，and have an inhibitory effect on the HPGA axis.