金钱草煎汤煮制雷公藤的炮制减毒工艺优选及其机制  

Optimization of Processing and Detoxification Process for Leigongteng(Tripterygium wilfordii)Boiled in Jinqiancao(Lysimachia christinae)Decoction and Preliminary Exploration of Related Mechanism

作　　者：王彦嵋 王君明[1,2] 关月晨 宋玲玲 张月月[1] 弓明珠 李炳印 巫晓慧 WANG Yanmei;WANG Junming;GUAN Yuechen;SONG Lingling;ZHANG Yueyue;GONG Mingzhu;LI Bingyin;WU Xiaohui(College of Pharmacy,Henan University of Chinese Medicine,Zhengzhou 450046,Henan,China;Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases by Henan&Education Ministry of P.R.China,Henan University of Chinese Medicine,Zhengzhou 450046,Henan,China)

机构地区：[1]河南中医药大学药学院,河南郑州450046 [2]河南中医药大学呼吸疾病中医药防治省部共建协同创新中心,河南郑州450046

出　　处：《中华中医药学刊》2023年第3期253-258,I0034,共7页Chinese Archives of Traditional Chinese Medicine

基　　金：河南省高校科技创新人才支持计划项目(16HASTIT032);河南中医药大学科技创新人才支持计划(2015XCXRC01)。

摘　　要：目的研究旨在优选金钱草煎汤煮制雷公藤的最佳炮制减毒工艺,并基于核因子κB(nuclear factor kappa-B,NF-κB)信号通路初步探讨炮制减毒的分子机制。方法以雷公藤与金钱草的质量比(因素A)、金钱草煎汤煮制液体积(因素B)和煮制温度(因素C)为考察因素,并设立清水煮制雷公藤炮制品作为对照组。在此基础上,将雷公藤生品及以上各炮制品分别以2 g·kg^(-1)(以雷公藤生药计)的剂量灌服小鼠21 d,末次给药后24 h取血和肝组织。采用试剂盒检测小鼠血清谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)的水平并结合苏木素-伊红染色法对肝组织病理切片分析优选金钱草煎汤煮制雷公藤的炮制减毒工艺,并进一步采用蛋白免疫印迹法和酶联免疫吸附法通过对肝NF-κB及其下游肿瘤坏死因子-α(tumor necrosis factor-alpha,TNF-α)、白细胞介素-1β(interleukin-1beta,IL-1β)水平的检测分析,探讨其炮制减毒机制。结果雷公藤生品给药致使小鼠血清ALT、AST水平显著升高,并伴有肝细胞变性及炎细胞浸润,而经炮制品干预后均在不同程度上抑制了雷公藤诱导的生化指标及病理异常情况,且以A3B2C1逆转率最高;此外,雷公藤给药还显著上调了肝脏p-NF-κB p65蛋白表达水平及炎症因子TNF-α、IL-1β水平,而不同炮制品干预后可逆转,其中以A3B2C1逆转率最高。结论金钱草煎汤煮制雷公藤炮制减毒的最佳工艺为A3B2C1,即“雷公藤与金钱草质量比为60∶7.5,取37.5 mL的金钱草煎汤煮制液母液稀释至200 mL,煮制温度为110℃”,其减毒机制可能与抑制主要毒性靶器官肝NF-κB介导的炎症反应有关。Objective The purpose of this study was to optimize the best processing and attenuating technology of Leigongteng(Tripterygium wilfordii)boiled in Jinqiancao(Lysimachia christinae)decoction and preliminarily explore the molecular mechanism of processing and attenuating based on the nuclear factor kappa-B(NF-κB)signaling pathway.Methods The quality ratio of Leigongteng(Tripterygium wilfordii)to Jinqiancao(Lysimachia christinae)(Factor A),the volume of Jinqiancao(Lysimachia christinae)water decoction(Factor B)and the decocting temperature(Factor C)were used as the investigating factors.Then considering the influence of heat and other factors,it was also necessary to prepare Leigongteng(Tripterygium wilfordii)boiled in water as a control group.On this basis,the raw products and the processed products of Leigongteng(Tripterygium wilfordii)were respectively intragastrically administered to mice at a dose of 2 g·kg^(-1)for 21 d,and blood and liver tissues were taken out 24 h after the last administration.The serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)of mice were detected by kits,and the pathological sections of liver tissue were analyzed by hematoxylin-eosin staining to optimize the processing and attenuating technology of Leigongteng(Tripterygium wilfordii)boiled with Jinqiancao(Lysimachia christinae).Then,the levels of NF-κB and its downstream tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)were further detected by western blotting and enzyme-linked immunosorbent assay to analyze and explore the mechanism of its processing and attenuation.Results Leigongteng(Tripterygium wilfordii)administration induced a significant increase in serum levels of ALT and AST in mice,accompanied by hepatocyte degeneration and inflammatory cell infiltration.Fortunately,after the intervention of the processed products,the biochemical indexes and pathological abnormalities induced by Leigongteng(Tripterygium wilfordii)were inhibited to varying degrees,and the reversal rate of A3B2C1 was t

关 键 词：雷公藤 肝毒性 金钱草煎汤煮制雷公藤 正交试验设计 炮制工艺 炎症 

分 类 号：R283.3[医药卫生—中药学]