木犀草素通过降低CYP2E1表达对重症急性胰腺炎大鼠相关急性肝损伤的保护作用研究

Study on Protective Effect of Luteolin on Rats with Severe Acute Pancreatitis-Related Acute Liver Injury by Reducing CYP2E1 Expressions

目的:探讨木犀草素通过降低细胞色素P4502E1(CYP2E1)表达对重症急性胰腺炎(SAP)大鼠相关急性肝损伤的保护作用。方法:将50只雄性SD大鼠分为假手术组、模型组和木犀草素低、中、高剂量组,每组10只。除假手术组外,用5%牛磺胆酸钠按1.5 mL/kg的剂量以0.2 mL/min的均匀速度注射到胆管内制备SAP模型,假手术组给予等量生理盐水。模型制备前2 h木犀草素低、中、高剂量组分别腹腔注射10.0 mg/kg、20.0 mg/kg、40.0 mg/kg的木犀草素,假手术组和模型组给予等量生理盐水。模型制备24 h后心脏取血,检测大鼠血清淀粉酶(AMY)、丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)水平;ELISA法检测血清中肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)水平;检测肝组织中谷胱甘肽(GSH)、活性氧(ROS)、CYP2E1、高迁移率族蛋白1(HMGB1)、Toll样受体4(TLR-4)、磷酸化白细胞介素-1受体相关激酶1(p-IRAK1)抗体水平。结果:与假手术组比较,模型组大鼠血清中AMY、ALT、AST、TNF-α、IL-1β水平及肝组织中ROS、CYP2E1、HMGB1、TLR-4、p-IRAK1蛋白表达水平均升高(P<0.05),肝组织中GSH水平降低(P<0.05);与模型组比较,木犀草素各剂量组大鼠血清中AMY、ALT、AST、TNF-α、IL-1β水平及肝组织中ROS、CYP2E1、HMGB1、TLR-4、p-IRAK1蛋白表达水平均降低(P<0.05),肝组织中GSH水平升高(P<0.05),且呈剂量依赖性(P<0.05)。结论:木犀草素可以减轻SAP相关急性肝损伤,其机制可能与下调CYP2E1的表达及抑制HMGB1-TLR-4信号通路有关。

Objective:To investigate the protective effect of luteolin on rats with severe acute pancreatitis(SAP)-related acute liver injury by reducing the expressions of cytochrome P4502 E1(CYP2 E1).Methods:A total of 50 male SD rats were divided into the sham operation group,the model group and the luteolin groups with low,medium and high dose respectively,10 rats in each group.Except for the sham operation group,the other groups were given the injection of 5%sodium taurocholate into the bile duct with a dose of 1.5 mL/kg at a uniform speed of 0.2 mL/min to prepare the SAP models,and the sham operation group was given the same amount of saline.Two hours before the model preparation,the luteolin groups with low,medium and high dose were intraperitoneally injected with 10.0 mg/kg,20.0 mg/kg and 40.0 mg/kg of luteolin respectively;the sham operation group and the model group were given the same amount of saline.After 24 hours of model preparation,blood samples were collected from the heart to detect the levels of amylase(AMY),alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in serum;the levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)were detected by ELISA method;the levels of glutathione(GSH),reactive oxygen species(ROS),CYP2 E1,high mobility group box protein 1(HMGB1),toll-like receptor 4(TLR-4),and the antibody of phosphated interleukin-1 receptor-associated kinase 1(p-IRAK1)in the liver tissue were detected.Results:Compared with those in the sham operation group,the levels of AMY,ALT,AST,TNF-α,IL-1βin serum and the expression levels of ROS,CYP2 E1,HMGB1,TLR-4 and p-IRAK1 in the liver tissue in the model group were all increased(P<0.05),and GSH level in the liver tissue was decreased(P<0.05).Compared with those in the model group,the levels of AMY,ALT,AST,TNF-α,IL-1βin serum as well as the expression levels of ROS,CYP2 E1,HMGB1,TLR-4 and p-IRAK1 in the liver tissue in the luteolin groups with low,medium and high dose were all decreased(P<0.05),and GSH level in the liver tissue was increased(P<0.05),which showed a dose-dependent manner(P<0.05).Conclusion:Luteosin can reduce SAP-related acute liver injury,and its mechanism may be related to the down-regulation of CYP2 E1 expressions and the inhibition of HMGB1-TLR-4 signaling pathway.