水飞蓟宾改善INH/RFP致肝损伤模型中原卟啉Ⅸ的作用及其机制  

Study on the mechanism of Silybin improving the accumulation of protoporphyrin IX in INH/RFP induced liver injury model

作　　者：李思云 麦穗晴 林迎瑶 张佳娜 何丽霞[1] 张雪婉 李书贤 汤学谦 邹晓婷 王聪 许雪飞 LI Si-yun;MAI Sui-qing;LIN Ying-yao;ZHANG Jia-na;HE Li-xia;ZHANG Xue-wan;LI Shu-xian;TANG Xue-qian;ZOU Xiao-ting;WANG Cong;XU Xue-fei(The Second Clinical College of Guangzhou Medical University,Guangzhou 5102601 China;The Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510180,China)

机构地区：[1]广州医科大学第二临床学院,广东广州510260 [2]广州医科大学附属第二医院,广东广州510260

出　　处：《解剖学研究》2022年第2期151-155,共5页Anatomy Research

基　　金：广东省中医药局科研项目(20202108);省级大学生创新创业训练计划(2020A022)。

摘　　要：目的初步探讨水飞蓟宾能否通过激动Nrf2/ARE信号通路,激活下游血红素合成中间体血卟啉Ⅸ(PPIX)代谢酶亚铁鳌合酶(FECH)的表达,从而改善异烟肼和(或)利福平(INH/RFP)引起的肝损伤。方法选用SPF级ICR小鼠随机分成5组:对照组(CON组)、模型组(IR组)、水飞蓟宾干预组(SY50组、SY100组、SY200组),干预组及IR组根据体质量给予75 mg/kgINH+150 mg/kgRFP,干预组分别同时给予水飞蓟宾低剂量组50 mg(IR+SY50),中剂量100 mg(IR+SY100)以及高剂量200 mg(IR+SY200),对照组给予空白溶剂,连续给药14 d。第15天给予异氟烷吸入麻醉后摘眼球取血,离心后用于生化指标测定,取肝组织于冻存管-80℃保存,Western blot测定目标蛋白FECH、Nrf2的表达。结果与对照组相比,IR组均能够引起生化指标ALT、AST、TBil、DBil、ALP、TBA的显著变化,与模型组组相比,低剂量的水飞蓟宾干预组ALT、Tbil、TBA降低明显,差异有统计学意义(P<0.05),中剂量的水飞蓟宾能显著降低ALT、AST、Tbil、TBA,高剂量的水飞蓟宾能显著降低除ALP以外的所有指标,差异有统计学意义(P<0.05)。模型组FECH蛋白表达下调,而水飞蓟宾能显著上调这些蛋白的表达,差异有统计学意义(P<0.01),干预组无明显剂量依赖性。与CON组相比,模型IR组总蛋白Nrf2表达上调,而核蛋白表达明显下调,差异有统计学意义(P<0.01),与IR组相比,水飞蓟宾干预组中Nrf2总蛋白下调,而Nrf2核蛋白表达均上调,差异有统计学意义(P<0.01)。结论水飞蓟宾能够激活Nrf2入核,调控下游血卟啉Ⅸ代谢酶FECH的表达,治疗INH/RFP引起的肝损伤。Objective To explore whether Silibinin can activate the expression of hematoporphyrin IX(PPIX)metabolizing enzyme(ferrochelatase,FECH)through activating Nrf2/ARE signal pathway,so as to improve the liver injurycaused by INH/RFP.Methods SPF ICR mice were randomly divided into 5 groups:control group(CON group),modelgroup(IR group)and Silybin intervention group(SY50 group,SY100 group and SY200 group).The intervention group andIR group were given 75 mg/kg INH+150 mg/kg RFP according to body weight,the control group was given blank solvent,and the intervention group was given Silybin 50 mg(IR+SY50),100 mg(IR+SY100)and 200 mg(IR+SY200)respectivelyfor 14 days.On the 15th day,eyeball blood was taken after isoflurane inhalation anesthesia and centrifuged for biochemicalindex determination,liver tissue was taken and stored in a cryopreservation tube at-80℃,the expressions of target proteinsFECH and Nrf2 were determined by Western blot.Results Compared with the control group,the IR group could causesignificant changes in biochemical indexes ALT,AST,TBIL,DBIL,ALP and TBA.Compared with the model group,thealt,TBIL and TBA in the low-dose Silybin intervention group decreased significantly,The difference was statistically significant(P<0.05).Medium dose silybin could significantly reduce ALT,AST,TBIL and TBA,and high dose Silybin could significantly reduce all indexes except ALP(P<0.05).The expression of FECH were downregulated in the IR group,while Silybin could significantly upregulate the expression of it(P<0.05).There was no significant dose dependence in the intervention group.Compared with the control group,the total protein Nrf2 expression was upregulated and the nuclear protein expression was significantly down regulated in the model IR group(P<0.01).Compared with the IR group,the total protein Nrf2 was down regulated and the nuclear protein Nrf2 expression was up regulated in the Silybin intervention group(P<0.01).Conclusion Silybin can activate Nrf2 into the nucleus,regulate the expression of downstream hematoporphyr

关 键 词：异烟肼 利福平 药物性肝损伤 Nrf2/ARE信号通路 水飞蓟宾 

分 类 号：R285.5[医药卫生—中药学]