何首乌中顺式(反式)-大黄素-大黄素二蒽酮肝保护活性研究  

Hepatoprotective activity of cis-(trans)-emodin-emodin dianthrone from Polygonum multiflorum Thunb.

作　　者：陈智伟 杨建波[2] 陈子涵 马双成[2] 孙华 CHEN Zhi-wei;YANG Jian-bo;CHEN Zi-han;MA Shuang-cheng;SUN Hua(State Key Laboratory of Bioactive Substance and Function of Natural Medicines,Institute of Materia Medica,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China;National Institutes for Food and Drug Control,Beijing 100050,China)

机构地区：[1]中国医学科学院、北京协和医学院药物研究所,天然药物活性物质与功能国家重点实验室,北京100050 [2]中国食品药品检定研究院,北京100050

出　　处：《药学学报》2023年第3期711-720,共10页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81973476);中国医学科学院医学与健康科技创新工程重大协同创新项目(2021-I2M-1-028,2021-I2M-1-029)。

摘　　要：顺式-大黄素-大黄素二蒽酮(化合物1)和反式-大黄素-大黄素二蒽酮(化合物2)均从何首乌的干燥块根中分离得到。在本研究中,首先评价了化合物1和2的混合样品(大黄素-大黄素二蒽酮)对刀豆蛋白A(concanavalin A,ConA)诱导ICR小鼠急性肝损伤的保护作用及可能的作用机制。结果表明,大黄素-大黄素二蒽酮在1 mg·kg^(-1)可显著降低肝损伤小鼠血清谷丙转氨酶(alanine aminotransferase,ALT)和谷草转氨酶(aspartate aminotransferase,AST)水平(P<0.05),改善肝组织病理损伤。1 mg·kg^(-1)大黄素-大黄素二蒽酮还能显著降低肝组织Bcl-2相关X蛋白(Bcl-2 assaciated X protein,Bax)mRNA水平,同时升高B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)mRNA表达水平(P<0.05)。进一步利用过氧化氢(H_(2)O_(2))诱导的肝细胞损伤模型评价化合物1和2对肝细胞损伤的保护活性。结果表明,化合物1和2可显著抑制H_(2)O_(2)诱导的肝细胞损伤,降低细胞培养上清转氨酶ALT、AST及碱性磷酸酶(alkaline phosphatase,ALP)、乳酸脱氢酶(lactate dehydrogenase,LDH)水平,提高细胞存活率。化合物1和2显著改善H_(2)O_(2)诱导的肝细胞氧化应激状态,0.5μmol·L^(-1)化合物1可显著提高肝细胞内超氧化物歧化酶(superoxide dismutase,SOD)的酶活力(P<0.01),0.5μmol·L^(-1)化合物2可显著降低细胞内活性氧(reactive oxygen species,ROS)含量、提高SOD酶活力和还原型谷胱甘肽(glutathione,GSH)的含量(P<0.01)。同时,0.5μmol·L^(-1)化合物1和2可通过提高Bcl-2/Bax蛋白表达比例(P<0.05)、降低剪切体半胱天冬酶3(cleaved caspase-3)和前体胱天蛋白酶3(pro caspase-3)的比例抑制肝细胞凋亡(P<0.05)。本研究表明,何首乌中大黄素-大黄素二蒽酮成分具有抗肝组织损伤活性,其中化合物1和2可通过抑制细胞凋亡和氧化应激发挥肝保护作用。本研究为常用量何首乌肝补益功效提供了重要的物质基础。本研究中所有动物实验在开展前经过The cis-emodin-emodin dianthrone(compound 1)and trans-emodin-emodin dianthrone(compound 2)were extracted from Polygonum multiflorum Thunb.The protective effect and mechanism of compound 1 and compound 2(emodin-emodin dianthrones)on acute liver injury induced by concanavalin A(ConA)in ICR mice was first investigated.The results indicated that emodin-emodin dianthrones at 1 mg·kg^(-1)significantly reduced serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)level(P<0.05).Emodin-emodin dianthrones also improved liver histopathological damage in liver-injured mice.The level of Bcl-2-associated X protein(Bax)mRNA in liver was significantly reduced by 1 mg·kg^(-1)of emodin-emodin dianthrones,while the level of B-cell lymphoma-2(Bcl-2)mRNA expression was significantly increased(P<0.05).The protective activity of compounds 1 and 2 against hepatocyte injury was further evaluated by hydrogen peroxide(H_(2)O_(2))-induced hepatocyte injury.Compounds 1 and 2 significantly inhibited H_(2)O_(2)-induced hepatocyte injury and reduced the levels of ALT,AST,alkaline phosphatase(ALP),and lactate dehydrogenase(LDH)in cell culture.Compounds 1and 2 also significantly improved the cell survival rate and decreased H_(2)O_(2)-induced oxidative stress in hepatocytes.Compound 1(0.5μmol·L^(-1))significantly increased the enzymatic activity of superoxide dismutase(SOD)in hepatocytes(P<0.01),and 0.5μmol·L^(-1)of compound 2 significantly decreased the intracellular reactive oxygen species(ROS),increased SOD enzyme activity,and glutathione(GSH)content(P<0.01).Compounds 1 and 2 at 0.5μmol·L^(-1)also inhibited hepatocyte apoptosis by increasing the protein expression ratio of Bcl-2/Bax(P<0.05)and decreasing the protein expression ratio of cleaved caspase-3 and pro caspase-3(P<0.05).This study indicates that the emodin-emodin dianthrones from Polygonum multiflorum Thunb.have liver-protective activity.Compounds 1 and 2 exerted hepatoprotective effects by inhibiting apoptosis and oxidative stress.The study provides an importa

关 键 词：何首乌 细胞凋亡 氧化应激 肝损伤 大黄素-大黄素二蒽酮 

分 类 号：R966[医药卫生—药理学]