摘要
研究五味子乙素(schisandrin B,Sch B)对顺铂致HK-2细胞氧化应激损伤的保护作用及其相关机制。体外培养人肾小管上皮细胞HK-2细胞,采用MTT和LDH法检测细胞毒性,荧光素DCFH-DA探针法测定细胞内ROS含量,Western blotting检测评价Nrf2核内水平,实时定量PCR检测NQO1、HO-1和GCLC mRNA表达水平。结果显示,经Sch B干预后,细胞活力明显升高,LDH漏出显著降低,并呈现剂量依赖效应。Sch B干预能显著抑制顺铂诱导的细胞内ROS水平升高。同时,研究发现Sch B能显著诱导Nrf2核转位,其下游Ⅱ相解毒酶基因和抗氧化酶基因NQO1、GCLC和HO-1 mRNA表达水平也显著增加。本研究表明,Sch B能通过激活Nrf2/ARE通路,减轻顺铂致肾细胞的氧化应激损伤作用。
This study is to investigate the protection effect of schisandrin B （Sch B） against oxidation stress of HK-2 cells induced by cisplatin and the mechanisms involved. HK-2 cells were cultured and divided into different groups： solvent control group, cisplatin exposure group, positive group, Sch B treatment group. Cell viability and toxicity were evaluated by MTT and LDH assay. GSH level and SOD enzymes activities were also measured. DCFH-DA as fluorescence probe was used to detect ROS level by fluorescence microplate reader. Nrf2 translocation was detected by Western blotting. Real time Q-PCR was used to detect expressions ofNQO1, HO-land GCLC mRNA level. The results showed that Sch B could significantly inhibit the decline of cell viability induced by cisplatin treatment （P 〈 0.05） and the protective effect was in a dose dependent manner. Furthermore, Sch B treatment significantly inhibited the increase of ROS level induced by cisplatin and reversed the decrease of GSH level （P 〈 0.05）. When Sch B concentration was up to 5 μmol.L-1, SOD enzyme activities were also enhanced significantly compared with that of the cisplatin group （P 〈 0.05）. It was shown that Sch B could cause nuclear accumulation of Nrf2 in association with downstream activation of Nrf2 mediated oxidative response genes such as GCLC, NQO1 and HO-1. These results suggested Sch B could protect against the oxidative damage of HK-2 cells induced by cisplatin via the activation of Nrf2/ARE signal pathway.
出处
《药学学报》
CAS
CSCD
北大核心
2012年第11期1434-1439,共6页
Acta Pharmaceutica Sinica
基金
国家自然科学基金资助项目(81102886)
中山大学实验室开放基金(KF201143)
