马兜铃酸I激活大鼠肾脏p38 MAPK通路并导致肾细胞凋亡的研究  

Study of Aristolochic Acid I in Activating p38 MAPK Pathway and Leading to Renal-cell Apoptosis in Rat's Renal Tissue

作　　者：张良[1] 杨召聪[1] 顾亚琴[1] 钱知知 周玲玲[1] 

机构地区：[1]南京中医药大学药学院,江苏省药效与安全性评价重点实验室,江苏南京210023

出　　处：《中药新药与临床药理》2015年第5期576-581,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：江苏省科技厅自然科学基金面上项目(BK2012852);江苏省高校优势学科建设工程资助项目(2011XYZ4-003);江苏省2013年度普通高校研究生科研创新计划项目(CXLX13-594)

摘　　要：目的探讨马兜铃酸I(aristolochic acid I,AAI)诱导马兜铃酸肾病可能的病变机制,分析AAI对p38丝裂原活化蛋白激酶(p38 MAPK)通路及其下游相关凋亡通路的影响。方法 SD雄性大鼠按体质量随机分为溶媒对照组,AAI高、低剂量(9.0,2.25 mg·kg-1)组,连续给药14 d。用生化试剂盒测定肾组织中过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、丙二醛(MDA);用蛋白免疫印迹(Western blot)法分析p38、c-jun氨基末端激酶(JNK)及其磷酸化水平和凋亡相关蛋白水平;用TUNEL染色法观察AAI对肾脏组织细胞凋亡的影响。结果与溶媒对照组比较,AAI组肾组织中MDA明显含量升高(P<0.05,P<0.001),SOD及CAT活力明显降低(P<0.01,P<0.001);总p38(t-p38)、磷酸化p38(p-p38)蛋白表达水平升高(P<0.01,P<0.001);总JNK(t-JNK)、磷酸化JNK(p-JNK)蛋白表达水平无明显变化;Caspase-3、Caspase-9、Bax等凋亡相关蛋白表达水平升高(P<0.05),Bax/Bcl-2比值升高(P<0.05);肾组织中凋亡小体个数明显增多(P<0.01,P<0.001)。结论 AAI可能通过激活p38 MAKP通路导致肾脏组织细胞凋亡而产生肾毒性效应。Objective To investigate the mechanisms of aristolochic acid nephropathy induced by aristolochic acid I （AAI） and to investigate the effect of AAI on the MAPK signaling pathway and related down-stream apoptosis pathways in rat＇s renal tissue. Methods Male SD rats were randomly divided into solvent control group, low-dose AAI group（2.25 mg·kg-1） and high-dose AAI group（9.0 mg·kg-1）. Rats were intraperitoneally injected with AAI once daily for 14 continuous days. On medication day 14, kidney tissues were isolated for the determination of catalase （CAT）, superoxide dismutase（SOD）, and malondialdehyde（MDA） with biochemical kits; Western blot was used to analyze the levels of t-p38, t-JNK and their levels of phosphorylation as well as other proteins associated with apoptosis; the apoptosis level of renal cells were examined by TUNEL assay. Results Compared with the solvent control group, the level of MDA in the two AAI groups were increased significantly（P 〈 0.05, P 〈 0.001） and the activities of CAT and SOD were decreased （P 〈 0.01, P 〈 0.001） ; the expression levels of t-p38 and p-p38 were increased（P 〈 0.01, P 〈 0.001） , but the levels of t-JNK and p-JNK stayed unchanged; apoptosis-associated proteins such as Caspase3, Caspase9 and Bax were up-regulated（P 〈 0.05）, Bax/Bcl-2 ratio was enhanced（P 〈 0.05）, and the number of renal apoptotic bodies was increased significantly（P 〈 0.01, P 〈 0.001 ）. Conclusion It is indicated that AAI could induce the injury of kidney by activating the p38 MAKP signaling pathway leading to renal-cell apoptosis in renal tissues of rats

关 键 词：马兜铃酸I 肾毒性 P38 MAKP通路 凋亡 

分 类 号：R285.5[医药卫生—中药学]