基于Keap1/Nrf2信号通路针灸拮抗DDP肾损伤小鼠的作用机制

Mechanism of Acupuncture and Moxibustion Antagonizing DDP Renal Injury Mice Based on Keap1/Nrf2 Signal Pathway

目的以顺铂(DDP)诱导肾损伤模型小鼠,通过观察针灸对肾脏组织中Kelch样环氧氯丙烷相关蛋白1(Keap1)/核因子E2相关因子2(Nrf2)信号通路的影响,阐释针灸改善DDP所致肾损伤的保护机制。方法选用清洁级雄性KM小鼠40只,随机分为4组,每组10只。在干预开始前1天,对空白组外的3组小鼠按体质量腹腔注射顺铂10 mg·kg^(-1),空白组注射同等剂量的0.9%NaCl溶液,24 h后模型即成。针刺组与艾灸组均选取“大椎”“肝俞”“肾俞”“足三里”,分别进行针刺与艾灸,每天1次,连续5天,其余2组陪同固定不予干预。禁食1天后取材,运用ELISA法检测血清中尿素氮(BUN)、肌酐(Scr)、半胱氨酸蛋白酶抑制剂(CysC)、中性粒细胞明胶酶相关脂质运载蛋白(NGAL)含量及肾脏组织中Keap1和Nrf2的含量;Western blot法及Real-time PCR法检测各组小鼠肾脏组织中Keap1和Nrf2蛋白表达及基因转录情况。结果与空白组比,模型组肾脏组织中Keap1含量、蛋白表达及mRNA相对表达均增多(P<0.05),Nrf2含量及蛋白表达减少(P<0.05),Nrf2 mRNA相对表达增多;与模型组比,针刺和艾灸组小鼠肾脏组织中Keap1含量、蛋白表达以及mRNA相对表达均减少(P<0.05);Nrf2含量、蛋白表达以及mRNA相对表达均增多(P<0.05)。结论针灸可以改善DDP肾损伤模型小鼠的肾功能,提升其抗氧化应激能力,从而改善DDP化疗所致的肾损伤,其作用机制可能与Keap1/Nrf2信号通路有关。

Objective To observe the impacts of acupuncture and moxibustion on kelch like epichlorohydrin related protein 1(Keap1)and nuclear factor E2 related factor 2(Nrf2)signal pathway in kidney tissue of cisplatin(DDP)-induced kidney injury model mice,and to explain the protective mechanism of acupuncture and moxibustion on improving kidney injury caused by DDP.Methods Forty SPF male KM rats were randomly divided into 4 groups,with 10 rats in each group.One day before the start of treatment,the three groups of mice outside the blank group were intraperitoneally injected with cisplatin 10 mg·kg^(-1)according to their body weight,and the blank group was injected with the same dose of 0.9%NaCl solution.The model was established 24 hours later.Both acupuncture group and moxibustion group selected"Dazhui"(GV14),"Ganshu"(BL18),"Shenshu"(BL23)and"Zusanli(Housanli)"(ST36)for acupuncture and moxibustion respectively,once a day for 5 days.The other two groups were fixed every day without treatment.After fasting for 1 day,the contents of BUN,Scr,CysC and NGAL in serum and Keap1 and Nrf2 in renal tissue were detected by ELISA;Western blot and Real-time PCR were used to detect the protein expression and gene transcription of Keap1 and Nrf2 in the kidney tissue of mice in each group.Results Compared with the blank group,the content of Keap1 protein,protein expression and relative expression of mRNA in the model group increased(P<0.05),the content of Nrf2 protein,protein expression decreased(P<0.05),Nrf2 relative expression of mRNA increased(P<0.05);Compared with the model group,the content of Keap1 protein,the expression of Keap1 protein and the relative expression of Keap1 mRNA in the kidney of mice in the acupuncture and moxibustion groups decreased(P<0.05);Nrf2 protein content,protein expression and relative mRNA expression increased(P<0.05).Conclusion Acupuncture and moxibustion can improve the renal function of DDP renal injury model mice and enhance their antioxidant stress ability,so as to improve the renal injury caused by DDP chemotherapy.Its mechanism may be related to Keap1/Nrf2 signal pathway.