姜黄素对酒精性肝损伤大鼠CYP3A的影响及机制探讨  

The effect of curcumin on CYP3A in rats with alcoholic liver injury and its mechanism

作　　者：陈洁[1] 刘斌[2] 袁桥玉[1] CHEN Jie;LIU Bin;YUAN Qiao-yu(College ofBioengineering,Wuhan Polytechnic,Wuhan 430074,China;Department ofPharmacy,Hubei CancerHospital)

机构地区：[1]武汉职业技术学院生物工程学院,430074 [2]湖北省肿瘤医院药学部

出　　处：《天津医药》2021年第12期1276-1281,共6页Tianjin Medical Journal

摘　　要：目的探究姜黄素对酒精性肝损伤(ALD)大鼠细胞色素P450 3A(CYP3A)的影响及其机制。方法 60只建模成功的ALD大鼠按随机数字表法分为模型组,姜黄素低、中、高剂量组(分别灌胃40、80、160 mg/kg姜黄素)及阳性对照组(腹腔注射200 mg/kg腺苷蛋氨酸),每组12只;对照组12只正常饲养,灌胃等体积生理盐水,连续6周。全自动生化分析仪检测血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和碱性磷酸酶(ALP)水平;苏木精-伊红(HE)染色观察肝脏形态变化;荧光定量聚合酶链反应(q PCR)检测肝脏组织中孕烷X受体(PXR)、组成型雄甾烷受体(CAR)、CYP3A25 mRNA水平;Western blot检测肝脏组织中PXR、CAR蛋白水平。以大鼠原代肝细胞为研究对象,分别用0、100、200、300、400、500 mmol/L乙醇培养细胞,取细胞增殖抑制率约为50%时的乙醇浓度进行下一步实验;0、0.5、1.0、2.0、4.0、8.0、16.0μmol/L姜黄素培养细胞,取最适姜黄素浓度进行后续研究。实验分为对照组、乙醇组、姜黄素组、姜黄素+siRNA-NC组及姜黄素+siRNA-CYP3A25组。CCK-8检测细胞增殖情况;qPCR检测细胞中CYP3A25 mRNA水平;Western blot检测细胞中PXR、CAR蛋白水平。结果动物实验:与对照组相比,模型组血清中ALT、AST、ALP水平升高(P<0.05);与模型组相比,姜黄素低剂量组血清中ALT、ALP水平降低,肝脏组织中PXR、CAR mRNA和蛋白,CYP3A25 mRNA水平升高(P<0.05),姜黄素中、高剂量组血清中ALT、AST、ALP水平降低,肝脏组织中PXR、CAR mRNA和蛋白,CYP3A25 mRNA水平升高(P<0.05);随着剂量升高,各指标逐渐恢复。细胞实验:与对照组相比,乙醇组细胞增殖抑制率升高(P<0.05),细胞中PXR、CAR蛋白水平降低(P<0.05);与乙醇组相比,姜黄素组细胞增殖抑制率降低(P<0.05),细胞中CYP3A25 mRNA,PXR、CAR蛋白水平升高(P<0.05);与姜黄素组相比,姜黄素+siRNA-CYP3A25组细胞增殖抑制率升高(P<0.05),细胞中CYP3A25 mRNA,PXR、CAR蛋白水�Objective To investigate the effects of curcumin on cytochrome P450 3A(CYP3A) in rats with alcoholic liver injury(ALD) and its mechanism.Methods Sixty ALD rats with successful modeling were randomly divided into the model group,the curcumin low,medium and high dose groups(gavage 40,80 and 160 mg/kg curcumin respectively) and the positive control group(200 mg/kg ademetionine intraperitoneally),with 12 rats in each group.The control group(n=12) was fed normally and given intragastric administration of equal volume of normal saline,for 6 consecutive weeks.The serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST) and alkaline phosphatase(ALP) were detected by automatic biochemical analyzer.The morphological changes of liver were detected by hematoxylin-eosin(HE) staining.The mRNA levels of pregnane X receptor(PXR),constitutive androstane receptor(CAR) and CYP3A25 were detected by quantitative real-time PCR(qPCR).The protein levels of PXR and CAR were detected by Western blot assay.Rat primary hepatocytes were used to culture with 0,100,200,300,400 and 500 mmol/L alcohol respectively,and the alcohol concentration was taken when the cell proliferation inhibition rate was about 50% for the next experiment.0,0.5,1.0,2.0,4.0,8.0 and 16.0 μmol/L curcumin cultured cells respectively,and the most suitable curcumin concentration was used for the next study.The experiments were divided into the control group,the alcohol group,the curcumin group,the curcumin+siRNA-NC group and the curcumin+siRNA-CYP3A25 group.The cell proliferation was detected by CCK-8.The level of CYP3A25 mRNA in cells was detected by qPCR.The levels of PXR and CAR protein in cells were detected by Western blot assay.Results In the rat experiment:compared with the control group,the serum levels of ALT,AST and ALP increased in the model group(P<0.05).Compared with the model group,the serum levels of ALT and ALP decreased in the curcumin low dose group,and the levels of PXR,CAR mRNA and protein,CYP3A25 mRNA in liver tissue increased(P<0

关 键 词：姜黄素 化学性与药物性肝损伤 肝疾病 酒精性 细胞色素P-450 CYP3A 孤儿核受体 孕烷X受体 组成型雄甾烷受体 

分 类 号：R575.5[医药卫生—消化系统]