松油烯-4-醇调节SIRT1/Nrf2信号抑制慢性肾病血管氧化应激损伤

Terpinen-4-ol regulates SIRT1/Nrf2 signaling to inhibit vascular oxidative stress injury in chronic kidney disease

目的探讨松油烯-4-醇(T4O)对慢性肾脏疾病(CKD)模型小鼠血管氧化应激损伤的作用及信号机制。方法采用高磷饲料联合腺嘌呤制备CKD小鼠模型,正常对照组给予等体积生理盐水灌胃。通过尾静脉注射慢病毒SIRT1 RNAi,建立SIRT1体内低表达的CKD小鼠模型用于信号机制研究。给药组以T4O低、高剂量(10 mg/kg和20 mg/kg)连续灌胃给药6周。收集小鼠血清检测尿素氮(BUN)和肌酐(CRE)水平,采用HE染色观察小鼠胸主动脉和肾脏组织形态,试剂盒检测血管组织丙二醛(MDA)及超氧化物歧化酶(SOD)含量,免疫荧光检测血管组织ROS水平;Western blot法检测Nrf2、HO-1、NQO-1及SIRT1的表达。结果T4O可降低CKD小鼠血清BUN和CRE水平,改善肾脏病理损伤和主动脉血管形态结构,降低血管组织MDA含量,增加SOD含量,降低ROS水平;T4O干预能够促进Nrf2核易位及上调HO-1、NQO-1以及SIRT1的蛋白表达。分子机制研究表明,LV-SIRT1 RNAi+T4O+CKD组能够抑制T4O对CKD诱导的MDA和SOD含量的影响,部分抵消T4O上调Nrf2核易位以及SIRT1、HO-1、NQO-1的蛋白表达水平的作用。结论T4O对CKD小鼠胸主动脉氧化应激损伤具有保护作用,其分子信号机制可能与调节SIRT1/Nrf2级联信号有关。

Objective To investigate the effect and signaling mechanism of terpinen-4-ol(T4O)on vascular oxidative stress injury in mice with chronic kidney disease(CKD).Methods A CKD mice model was prepared using high phosphorus diet combined with adenine,and the normal group was given an equal volume of saline gavage.The CKD model with low expression of SIRT1 in vivo was established by tail vein injection of lentiviral SIRT1 RNAi for the study of signaling mechanism.The administration groups were given T4O at low and high doses(10 mg/kg and 20 mg/kg)for 6 weeks by continuous gavage.Serum was collected to detect urea nitrogen(BUN)and creatinine(CRE)levels,and HE staining was used to observe the morphology of blood vessels in the thoracic aorta of mice expression.Results T4O reduced serum BUN and CRE levels in CKD mice to improve renal function,improved kidney and thoracic aortic vascular morphology,reduced vascular tissue MDA content,increased SOD content,and reduced ROS levels;T4O intervention promoted Nrf2 nuclear translocation and upregulated HO-1,NQO-1 and SIRT1 protein expression;LV-SIRT1 RNAi+T4O group was able to inhibit the effect of T4O on CKD-induced MDA and SOD levels,partially counteracting the effect of T4O in upregulating Nrf2 nuclear translocation and the protein expression levels of SIRT1,HO-1 and NQO-1.Conclusion T4O has a protective effect against oxidative stress injury in the thoracic aorta of CKD mice,and its molecular signaling mechanism may be related to the level of drug-regulated SIRT1/Nrf2 cascade signaling.