网络药理学结合动物实验初步探究火把花根片的睾丸毒性  

Preliminary study on testicular toxicity of huobahuagen hablets by network pharmacology and animal experiment

作　　者：杜伟[1] 张莉[1] 宋小仙[1] 陈慧[1] 梅小利[1] 王天文[1] DU Wei;ZHANG Li;SONG Xiao-xian;CHEN Hui;MEI Xiao-li;WANG Tian-wen(Chongqing Academy of Chinese Materia Medica,Chongqing 400065,China)

机构地区：[1]重庆市中药研究院,重庆400065

出　　处：《毒理学杂志》2022年第5期408-413,共6页Journal of Toxicology

基　　金：重庆市科技局项目(cstc2017jxjl-jbky120028)。

摘　　要：目的 通过网络药理寻找火把花根片(HBHGH)脱靶靶点,并进行动物实验验证火把花根片睾丸毒性分子机制。方法 运用维普网、中药系统药理疗数据库与分析平台和人类基因组注释数据库等数据库,采用网络药理学方法分析火把花根片脱靶靶点及通路;将雄性KM种小鼠随机分为2组,即药物组和溶媒对照组,分别灌胃火把花根片1.0 g/kg·bw及等体积纯化水(20 ml/kg·bw),6个月后,采集睾丸样本,分别称量睾丸重量、计算睾丸相对指数、定性观察睾丸组织病理学变化、RT-PCR检测NR4A1、Bax、Bcl-2、3β-HSD、INSL3、Cyp17a1和Star mRNA表达。结果 火把花根片脱靶靶点包括AR、NR4A1、ESR2,NR4A1因生物功能及作用通路与睾丸毒性相关成为实验验证的靶点;火把花根片持续6个月重复给药诱导了小鼠睾丸毒性;组织病理学检查可见药物组曲精管凋亡、萎缩、间距改变,腔内皮改变,精原细胞和精母细胞数量改变;在检测的mRNA中,NR4A1、Bax、3β-HSD和INSL3较溶媒对照组升高,Bcl-2低表达,Bax/Bcl-2比值升高,其余Cyp17a1和Star变化不明显。结论 NR4A1是火把花根片诱导睾丸毒性的重要靶点,可能与调控凋亡(Bcl-2、Bax)和激素-精子生成(3β-HSD、INSL3)有关。Objective The target of Huobahuagen Hablets(HBHGH) was found by network pharmacology, and animal experiments were carried out to verify the molecular mechanism of HBHGH testicular toxicity. Methods The target and pathway of HBHGH were analyzed by using the database of CQVIP, TCMSP and GeneCards. The male KM mice were randomly divided into two groups, namely drug group and vehicle control group. The drug group was gavaged with HBHGH 1.0 g/kg·bw, and the vehicle control group was gavaged with equal volume of purified water(20 ml/kg·bw)for 6 months, the testis samples were collected, the weight of testis was weighed, the relative index of testis was calculated, the histopathological changes of testis were qualitatively observed, and the expressions of NR4 A1, Bax, Bcl-2, 3β-HSD, INSL3, Cyp17 a1 and Star mRNA were detected by RT-PCR. Results HBHGH off-targets include AR, NR4 A1 and ESR2, NR4 A1 has become the target of experimental verification due to its biological function and action pathway related to testicular toxicity. Repeated administration of HBHGH for 6 months induced testicular toxicity in mice. Histopathological examination showed that the apoptosis, atrophy, spacing changes of seminiferous tubules, lumen endothelium changes, the number of spermatogonia and spermatocytes changes. Among the detected mRNAs, NR4 A1, Bax, 3β-HSD and INSL3 were higher than those in the vehicle control group, the expression of Bcl-2 was lower, the ratio of Bax/Bcl-2 was higher, and the changes of Cyp17 a1 and Star were not significant. Conclusion NR4 A1 is an important target of testicular toxicity induced by HBHGH, which may be related to the regulation of apoptosis(Bcl-2 and Bax) and hormone-spermatogenesis(3β-HSD and INSL3).

关 键 词：火把花根片 KM小鼠 睾丸 网络药理 NR4A1 

分 类 号：R114[医药卫生—卫生毒理学] R285.5[医药卫生—公共卫生与预防医学] R99