基于靶器官铁死亡探讨绿豆汁炙黄药子的炮制减毒机制  

Exploring Detoxication Mechanism of Dioscoreae Bulbiferae Rhizoma Processed with Phaseoli Radiati Semen Juice Based on Target Organ Ferroptosis

作　　者：段雅倩 宋玲玲 张月月[1] 王君明[1] 刘鸣昊[2] 李亚敏[1] 李炳印 巫晓慧 王彦嵋 DUAN Yaqian;SONG Lingling;ZHANG Yueyue;WANG Junming;LIU Minghao;LI Yamin;LI Bingyin;WU Xiaohui;WANG Yanmei(Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases Co-constructed by Henan Province and Ministry of Education,Collaborative Innovation Center of Research and Development on the Whole Industry Chain of Yu-Yao,Henan Province,Henan University of Chinese Medicine,Zhengzhou 450046,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]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,豫药全产业链研发河南省协同创新中心,郑州450046 [2]河南中医药大学第一附属医院,郑州450000

出　　处：《中国实验方剂学杂志》2024年第10期70-76,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省重点研发与推广专项(182102310255)。

摘　　要：目的:探究绿豆汁炙黄药子的炮制减毒作用,并基于其主要毒性靶器官肝铁死亡探究其减毒机制。方法:60只雄性ICR小鼠随机分为空白组、生黄药子组、水炙黄药子组,绿豆汁炙黄药子1组(黄药子-绿豆10∶1,闷润40 min,130℃炒18 min)、绿豆汁炙黄药子2组(黄药子-绿豆10∶1,闷润80 min,100℃炒14 min)、绿豆汁炙黄药子3组(黄药子-绿豆20∶3,闷润40 min,160℃炒14 min)。生黄药子及各炮制品组均以3 g·kg^(-1)·d^(-1)的剂量分别灌胃对应的95%乙醇提取物药液,空白组灌胃等体积的0.5%羧甲基纤维素钠,1次/d,连续14 d。苏木素-伊红(HE)染色观察小鼠肝脏组织病理变化;生化检测小鼠血清谷氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)水平,以及肝组织丙二醛(MDA)、亚铁离子(Fe^(2+))、还原型谷胱甘肽(GSH)和超氧化物歧化酶(SOD)的水平;蛋白免疫印迹法检测铁关键蛋白铁蛋白重链1(FTH1)、谷胱甘肽过氧化物酶4(GPX4)的表达水平。结果:HE染色结果显示,空白组小鼠肝组织结构清晰,肝细胞形态正常;生黄药子组与水炙黄药子组小鼠肝细胞胞质疏松、空泡,病理性损伤明显;各绿豆汁炙黄药子组小鼠病理性损伤明显改善。与空白组比较,生黄药子组小鼠血清ALT、AST水平显著升高(P<0.01),肝组织MDA及Fe^(2+)水平显著升高(P<0.01),GSH、SOD水平显著降低(P<0.01),FTH1、GPX4蛋白表达显著下调(P<0.01);与生黄药子组比较,各绿豆汁炙黄药子组小鼠血清ALT、AST水平显著降低(P<0.01),肝组织MDA的水平明显降低(P<0.05,P<0.01),Fe^(2+)水平显著降低(P<0.01),GSH、SOD水平显著升高(P<0.01),FTH1、GPX4蛋白表达显著上调(P<0.01);与水炙黄药子组比较,各绿豆汁炙黄药子组小鼠血清AST水平明显降低(P<0.05,P<0.01),肝组织MDA水平显著降低(P<0.01),SOD水平明显升高(P<0.05,P<0.01),FTH1蛋白表达显著上调(P<0.01),绿豆汁炙黄药子2、3组小鼠血清ALT水平显著降低(P<0.01),肝Objective:To investigate the attenuating effect of Dioscoreae Bulbiferae Rhizoma(DBR)processed with Phaseoli Radiati Semen(PRS)juice,and explore the attenuating mechanism based on ferroptosis of the main toxic target organ.Method:Sixty male ICR mice were randomly divided into blank group,DBR group,water roasted DBR group(hereinafter referred to as water group),PRS juice-roasted DBR group 1(DBRPRS 10∶1,stuffy moistening for 40 min,stir-fried at 130℃ for 18 min,hereinafter referred to as group 1),PRS juice-roasted DBR group 2(DBR-PRS 10∶1,stuffy moistening for 80 min,stir-fried at 100℃ for 14 min,hereinafter referred to as group 2),PRS juice-roasted DBR group 3(DBR-PRS=20∶3,stuffy moistening for 40 min,stir-fried at 160℃ for 14 min,hereinafter referred to as group 3).The raw and processed groups of DBR were gavaged with their corresponding 95% ethanol extract at a dose of 3 g·kg^(-1)·d^(-1),while the blank group was gavaged with an equal volume of 0.5% sodium carboxymethyl cellulose,once a day for 14 consecutive days.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of mouse liver.Alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels in serum,as well as malondialdehyde(MDA),ferrous ions(Fe^(2+)),reduced glutathione(GSH)and superoxide dismutase(SOD)levels in liver tissue were detected by the biochemical detection.Western blot was used to detect the expression of iron key proteins such as ferritin heavy chain 1(FTH1)and glutathione peroxidase 4(GPX4).Result:HE staining results showed that the liver tissue structure of the blank group was clear,the morphology of hepatocytes was normal,the cytoplasms of hepatocytes in the DBR group and water group were loose and vacuolar,with obvious pathological damages,and the pathologic damages of mice in the group 1-3 were significantly improved.Compared with the blank group,the levels of ALT,AST,MDA and Fe^(2+)in mice from the DBR group were significantly increased(P<0.01),while GSH and SOD levels were significantly redu

关 键 词：黄药子 绿豆 药汁炙法 肝损伤 炮制减毒 铁死亡 氧化应激 

分 类 号：R22[医药卫生—中医基础理论] R28[医药卫生—中医学] R943.1R96