基于PI3K/Akt通路探讨仙茅改善去势大鼠肾阳虚的作用机制  

Curculiginis Rhizoma Ameliorates Kidney-Yang Deficiency in Castrated Rats via PI3K/Akt Signaling Pathway

作　　者：杨濛 王璐 张紫佳[1] 王峥涛[1] 徐颖 YANG Meng;WANG Lu;ZHANG Zijia;WANG Zhengtao;XU Ying(The MOE Key Laboratory for Standardization of Chinese Medicines and the Shanghai Key Laboratory for Compound Chinese Medicines,Institute of Chinese Materia Medica,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]上海中医药大学中药研究所,中药标准化教育部重点实验室,上海市复方中药重点实验室,上海201203 [2]中国中医科学院中药研究所,北京100700

出　　处：《中国实验方剂学杂志》2024年第20期46-53,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：上海市中医药三年行动计划建设项目[ZY(2018-2020)-CCCX-5002]。

摘　　要：目的:探讨仙茅改善雄性去势大鼠肾阳虚的药效作用及相关的作用机制。方法:通过网络药理学在药物和疾病相关数据库中筛选出仙茅的药物作用靶点及肾阳虚导致的男性生殖疾病的相关靶点,根据拓扑特征值筛选出关键靶标后,再进行基因本体(GO)功能及京都基因与基因组百科全书(KEGG)通路富集分析得出仙茅可能造成肾阳虚的相关通路,其次采用去势手术建立雄性大鼠肾阳虚模型,分为模型组、丙酸睾酮组(2 mg·kg^(-1))和仙茅低、中、高剂量组(0.14、0.28、0.56 g·kg^(-1)),另取8只健康雄性大鼠进行切口后立即缝合,用作假手术组,每天灌胃给药1次,连续6周。实验过程中,观察各组大鼠的一般情况,记录其体质量,实验结束后,计算附性器官脏器指数,苏木素-伊红(HE)染色观察精囊腺和前列腺病理变化;采用酶联免疫吸附测定法(ELISA)检测大鼠血清激素促黄体生成素(LH)、促卵泡激素(FSH)和睾酮(T)与血清细胞因子白细胞介素-6(IL-6)的水平,并根据网络药理学预测的结果在动物体内进行实验验证,采用实时荧光定量聚合酶链式反应和蛋白免疫印迹法进行相关mRNA与蛋白测定。结果:生物学功能富集在调节细胞对各种刺激反应、细胞内信号传导、代谢反应及生殖调控4个过表达模块,其中磷脂酰基醇3-激酶(PI3K)/蛋白激酶B(Akt)途径核心靶标富集较为显著,也是在调节细胞对各种刺激反应生物学过程富集程度最高的通路。体内实验结果显示,与模型组比较,仙茅给药剂量组大鼠体质量有增长趋势,仙茅低剂量组肾阳虚大鼠体质量增长量明显升高(P<0.05);仙茅各给药组大鼠精囊腺指数和附睾指数随着给药剂量的增加呈上升趋势,精囊腺指数没有显著性变化,仙茅高剂量组大鼠的附睾指数明显增加(P<0.05),仙茅各给药剂量组大鼠精囊腺上皮细胞增生有明显减轻,但空泡变性并没有明显改善;前列�Objective:To study the effect and mechanism of Curculiginis Rhizoma in ameliorating kidney-Yang deficiency in castrated rats.Method:The targets of Curculiginis Rhizoma and male reproductive diseases due to kidney-Yang deficiency were screened from relevant databases by network pharmacology,and key targets were screened out according to topological eigenvalues.After that,gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed to obtain the pathways of Curculiginis Rhizoma in treating kidney-Yang deficiency.The rat model of kidney-Yang deficiency was established by castration.The rats were assigned into model,testosterone propinate(2 mg·kg^(-1)),and low-,medium-,high-dose(0.14,0.28,0.56 g·kg^(-1),respectively)Curculiginis Rhizoma groups.Another 8 healthy male rats with first incising and then suturing were used as the sham group.The rats were administrated with corresponding agents by gavage once a day for 6 consecutive weeks.During the experiment,the general conditions of rats in each group were observed,and their body mass was recorded.At the end of the experiment,the indexes of accessory sex organs were calculated,and the pathological changes of the seminal vesicle glands and prostate glands were observed by hematoxylin-eosin(HE)staining.The serum levels of luteinizing hormone(LH),follicle-stimulating hormone(FSH),testosterone(T),and interleukin-6(IL-6)were determined by enzyme-linked immunosorbent assay.The results of the network pharmacological prediction were verified by animal experiments,and the expression levels of related genes and proteins were determined by real-time polymerase chain reaction and Western blot,respectively.Result:The biological functions enriched mainly involved four overexpression modules including regulation of cell responses to various stimuli,metabolic responses,regulation of intracellular signal transduction,and regulation of reproduction.Phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway was a significantly enric

关 键 词：仙茅 肾阳虚 网络药理学 磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(Akt)信号通路 

分 类 号：R2-0[医药卫生—中医学] R33R289R339.2