摘要
目的:研究复幼颗粒对达那唑诱导大鼠性早熟模型的作用和有关机制。方法:将21窝SD雌鼠随机分为正常组,模型组,亮丙瑞林组(0.1 g·kg^(-1))和复幼合剂组(37.9 g·kg^(-1)),复幼颗粒高、中、低剂量组(17.0,8.5,4.3 g·kg^(-1))。于5日龄时注射达那唑300μg建立性早熟模型,造模10 d后,开始给药干预。20日龄时开始检查阴道开口情况,通过苏木素-伊红(HE)染色观察性腺发育情况;通过放射性免疫检测血清中黄体生成素(LH),卵泡刺激素(FSH),雌二醇(E2)的含量;通过实时荧光定量聚合酶链式反应(Real-time PCR)检测下丘脑促性腺激素释放激素(GnRH),Kiss-1,G蛋白偶联受体54(GPR54)mRNA表达。通过免疫组化检测下丘脑GnRH细胞表达情况。结果:与正常组比较,模型组阴道开口显著提前,子宫,卵巢系数明显增加(P<0.05),表明达那唑诱导性早熟模型成功建立,GnRH,Kiss-1,GPR54表达也均明显增加(P<0.05),表明达那唑模型可以提前启动下丘脑-垂体-性腺(HPG)轴,从而诱发性早熟。与模型组比较,复幼颗粒中剂量组可显著延缓阴道开口时间(P<0.01),复幼颗粒高剂量可明显减轻子宫壁厚、子宫系数(P<0.05,P<0.01),中剂量可减轻卵巢系数和子宫壁厚(P<0.05),复幼颗粒高、中、低剂量均能明显下调血清激素E2,LH,FSH的含量(P<0.05,P<0.01),降低下丘脑GnRH,Kiss-1,GPR54 mRNA的表达(P<0.05,P<0.01),降低GnRH细胞的表达水平(P<0.05)。结论:复幼颗粒可通过调节Kiss-1/GPR54系统,下调GnRH的表达从而抑制HPG轴的启动达到治疗性早熟目的。
Objective:To study the effect and related mechanism of Fuyou granule on danazol-induced precocious puberty model in rats. Method: Totally 21 cages of SD female rats were randomly divided into normal group,model group,Leuprorelin(0.1 g·kg^(-1))and Fuyou mixture group(37.9 g·kg^(-1)),and high-dose,mid-dose and low dose Fuyou granule groups(17.0,8.5,4.3 g·kg^(-1)). Rats at 5 days of age were given a single subcutaneous injection of 300 μg danazol to establish the precocious puberty model. After 10 days of modeling,drug intervention was started. Vaginal opening was examined at the age of 20 days,and the gonadal development was observed by hematoxylin-eosin(HE)staining. The levels of serum luteinizing hormone(LH),folliclestimulating hormone(FSH)and estradiol(E2)were determined by radioimmunoassay. The mRNA expressions of hypothalamic gonadotropin releasing hormone(GnRH),Kiss-1,G protein-coupled receptor 54(GPR54)were detected by Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR), and the expression of GnRH cells in the hypothalamus was detected by immunohistochemistry. Result: Compared with the normal group,the vaginal opening of the model group was significantly earlier,and the uterus and ovarian coefficients were significantly increased(P<0.05),indicating that the danazol-induced precocious puberty model was successfully established. The expression levels of GnRH, Kiss-1, and GPR54 also increased significantly(P<0.05),indicating that the danazol model can activate the HPG axis in advance,thereby inducing precocious puberty. Compared with the model group,the mid-dose Fuyou granule group significantly delayed the time of vaginal opening(P<0.01),high-dose Fuyou granule group significantly reduced uterine wall thickness and uterine coefficient(P<0.05,P<0.01),mid-dose group reduced ovarian coefficient and uterine wall thickness(P<0.05). All the three dosage groups of Fuyou granule significantly reduced the content of serum hormones E2,LH and FSH(P<0.05,P<0.01),reduced the expression levels of hypothalamic GnRH,Kiss-1 and GPR54 mRNA(P<0.05),and decreased the expression of GnRH cells(P<0.05). Conclusion: Fuyou granule can achieve therapeutic precocity by regulating the Kiss-1/GPR54 system and down-regulating the expression of GnRH to inhibit the activation of the HPG axis.
作者
胡凯丽
孙文燕
李玉
张波
张萌
郭春彦
赵立波
王晓玲
畅洪昇
HU Kai-li;SUN Wen-yan;LI Yu;ZHANG Bo;ZHANG Meng;GUO Chun-yan;ZHAO Li-bo;WANG Xiao-ling;CHANG Hong-sheng(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100102,China;Beijing Children's Hospital,Capital Medical University,Beijing 100045,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2021年第3期39-46,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家“重大新药创制”科技重大专项(2018ZX09721003-002-001)
首都卫生发展科研专项(首发2018-2-2097)。
