基于CYP1A2、CYP3A4酶活性的苍耳子炮制减毒机制研究  

Study on the Attenuation Mechanism of Cang'erzi(Xanthii Fructus)Processing Based on CYP1A2 and CYP3A4 Enzyme Activity

作　　者：黄成[1] 周仁杰 黄保生 韩燕全[1] 朋汤义[1] 金传山[2] 吴德玲[2] HUANG Cheng;ZHOU Ren-jie;HUANG Bao-sheng;HAN Yan-quan;PENG Tang-yi;JIN Chuan-shan;WU De-ling(The First Affiliated Hospital of Anhui University of Traditional Chinese Medicine/The Third Level Laboratory of Traditional Chinese Medicine Preparation of the State Administration of Traditional Chinese Medicine,Hefei Anhui 230031,China;Anhui University of Traditional Chinese Medicine/Anhui Key Laboratory of Traditional Chinese Medicine/Anhui Engineering Technology Center of Modern Pharmaceutical Preparations,Hefei Anhui 230031,China)

机构地区：[1]安徽中医药大学第一附属医院/国家中医药管理局中药制剂三级实验室,安徽合肥230031 [2]安徽中医药大学/中药复方安徽省重点实验室/现代药物制剂安徽省工程技术中心,安徽合肥230031

出　　处：《中医药导报》2022年第5期79-83,共5页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：安徽省徽派炮制传承工作室项目(皖中医药发展秘〔2021〕10号)。

摘　　要：目的:研究苍耳子炮制前后对小鼠肝微粒体中CYP1A2和CYP3A4酶相对活性及肝细胞损害程度的影响。方法:49只小鼠按体质量随机分为正常组、苍耳子生品低剂量组、苍耳子生品中剂量组、苍耳子生品高剂量组、苍耳子炮制品低剂量组、苍耳子炮制品中剂量组、苍耳子炮制品高剂量组,每组7只。各给药组小鼠灌胃分别相应药物水煎液,正常组小鼠灌胃等体积的生理盐水,1次/d,连续10 d。运用Cooktail探针法将咖啡因(CYP1A2酶反应物)、咪达唑仑(CYP3A4酶反应物)、地西泮(内标物)和肝微粒体一同进行体外孵育,HPLC法测定肝微粒体中两种探针药物的消耗,计算酶的相对活性;ELISA法测定血清中天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)水平;肝组织HE染色切片进行病理观察。结果:苍耳子生品中、高剂量组和苍耳子炮制品高剂量组小鼠血清AST、ALT活性均高于正常组(P<0.05或P<0.01),苍耳子生品低剂量组、苍耳子炮制品中剂量组小鼠血清ALT活性均高于正常组(P<0.05),且在相同剂量下,苍耳子生品低、中、高剂量组小鼠血清AST、ALT活性均分别高于苍耳子炮制品低、中、高剂量组,但差异均无统计学意义(P>0.05);小鼠肝组织HE染色病理切片显示在相同剂量下苍耳子生品组比苍耳子炮制品组小鼠肝损伤情况更严重;苍耳子生品中、高剂量组和苍耳子炮制品中、高剂量组小鼠肝脏微粒体中CYP1A2活性均明显高于正常组(P<0.05或P<0.01),苍耳子生品高剂量组、苍耳子炮制品高剂量组小鼠肝脏微粒体中CYP3A4活性均明显高于正常组(P<0.05),同等剂量下,苍耳子生品低、中、高剂量组小鼠肝脏微粒体中CYP1A2、CYP3A4活性均分别高于苍耳子炮制品低、中、高剂量组,但差异均无统计学意义(P>0.05)。CYP1A2和CYP3A4酶相对活性与血清生化指标检测和肝组织病理切片观察结果有相同的趋势,即CYP1A2、CYP3A4酶相对活性越高,�Objective:To study the effects of Cang'erzi(Xanthii Fructus)on the relative activities of CYP1A2 and CYP3A4 enzymes in mouse liver microsomes and the degree of hepatocyte damage before and after processing.Methods:A total of 49 mice were randomly divided into the normal group,the raw low-dose group,the raw medium-dose group,the raw high-dose group,the processed low-dose group,the processed medium-dose group and the processed high-dose group according to body weight,with 7 mice in each group.The mice in each administration group were gavaged with corresponding drug decoction,and the mice in the normal group were gavaged with equal volume of normal saline,once daily,for 10 days.Cooktail probe method was used to incubate caffeine(CYP1A2 enzyme reactant),midazolam(CYP3A4 enzyme reactant),diazepam(internal standard)and liver microsomes in vitro.The consumption of two probe drugs in liver microsomes was determined by HPLC,and the relative activity of enzymes was calculated.The levels of AST and ALT in serum were measured by ELISA,and the pathological observation of liver tissue was performed by HE staining section.Results:The serum AST and ALT activities in the raw medium-dose group,the raw high-dose group and the processed high-dose group were higher than those in the normal group(P<0.05 or P<0.01),and the serum ALT activities in the raw low-dose group and the processed medium-dose group were higher than those in the normal group(P<0.05).At the same dose,the serum AST and ALT activities of the mice in the raw low,medium and high-dose groups of were higher than those in the processed low,medium and high-dose groups,but the difference was not statistically significant(P>0.05).The pathological sections of mouse liver tissue HE staining showed that the liver injury of the mice in raw group was more serious than that in processed group at the same dose.The activity of CYP1A2 in the liver microsomes of the mice in the raw medium,high-dose groups and the processed medium,high-dose groups were significantly higher than those

关 键 词：苍耳子 炮制减毒 肝损伤 CYP1A2酶 CYP3A4酶 相对活性 肝微粒体 小鼠 

分 类 号：R283.1[医药卫生—中药学]