九味楮实方对中枢性性早熟大鼠性激素及Kisspeptin蛋白表达的影响

Effects of Jiuwei Chushi Decoction on sex hormone and Kisspeptin protein expression of Central Precocious Puberty rats

目的研究九味楮实方对中枢性性早熟(central precocious puberty,CPP)模型大鼠血清性激素水平及下丘脑Kisspeptin蛋白表达的影响。方法以SD大鼠配种得到雌性仔鼠60只,按随机数字表法分为正常对照组、模型组、戈那瑞林组及九味楮实方高、中、低剂量组,每组10只。除正常对照组外,其余大鼠均皮下注射N-甲基-DL-天冬氨酸建立CPP模型,戈那瑞林组皮下注射戈那瑞林100μg/(kg·d),九味楮实方高、中、低剂量组分别灌胃九味楮实方浸膏剂5.75、2.87、1.43 ml/(kg·d)。采用ELISA法检测大鼠血清LH、FSH、雌二醇(E_(2))水平,Western blot法检测下丘脑促性腺激素释放激素(GnRH)、Kisspeptin蛋白表达情况。结果与模型组比较,九味楮实方低、中、高剂量组LH、E_(2)水平降低(P<0.05),FSH水平升高(P<0.05),九味楮实方中、高剂量组GnRH[(0.657±0.110)、(0.536±0.152)比(0.912±0.219)]、Kisspeptin[(0.508±0.035)、(0.347±0.073)比(0.659±0.030)]蛋白表达降低(P<0.05)。结论九味楮实方可干预CPP模型大鼠下丘脑Kisspeptin蛋白表达,抑制下丘脑GnRH表达,降低血清LH、E_(2)水平。

Objective To observe the effect of Jiuwei Chushi Decoction on sex hormone level of serum and Kisspeptin protein expression in hypothalamus of Central Precocious Puberty(CPP)rats model.Methods After sixty female rat were breeded from SD rats,they are randomly divided into six groups,which are normal control group,model group,Gonadorelin group,and high,medium and low dose group of Jiuwei Chushi Decoction,10 in each group.In addition to the normal control group,the other groups were subcutaneously injected with N-methyl-DL-aspartic acid to establish CPP model.Gonadorelin group was subcutaneously injected with Gonadorelin 100μg/(kg·d),and high,medium and low dose group of Jiuwei Chushi Decoction were intragastrated with Jiuwei Chushi Decoction extract at 5.75,2.87 and 1.43 ml/(kg·d)respectively.The levels of Luteinizing Hormone(LH),Follicle Stimulating Hormone(FSH),Estradiol(E_(2))were detected by ELISA,and the protein expressions of Gonadotropin-releasing Hormone(GnRH)and Kisspeptin in hypothalamus were detected by Western Blot.Results Compared with the model group,the levels of LH and E_(2) of the low,medium and high dose group of Jiuwei Chushi Decoction were significantly decreased(P<0.05),while the FSH level was significantly increased(P<0.05).The relative protein expression of GnRH(0.657±0.110,0.536±0.152 vs.0.912±0.219)and Kisspeptin(0.508±0.035,0.347±0.073 vs.0.659±0.030)in the medium and high dose group of Jiuwei Chushi Decoction was lower than that of model group.Conclusion Jiuwei Chushi Decoction could affect hypothalamic Kisspeptin protein expression,inhibite hypothalamic GnRH expression,and decrease LH and E_(2) levels in the serum of CPP rats.