摘要
目的探讨单羰基姜黄素类似物（MH21）对百草枯（paraquat，PQ）诱导人肾近曲小管上皮细胞（HK．2）细胞损伤的保护作用及机制。方法培养HK-2细胞，构建PQ诱导细胞毒性损伤模型并同时以L6H21干预，设阴性对照组、PQ组、PQ＋L6H21组、L6H21对照组。应用CCK-8、细胞流式检测HK-2细胞增殖及凋亡情况；RT-PCR法检测凋亡、炎症等相关因子mRNA表达；双抗体夹心酶联免疫吸附法（ELISA）、Westernblot检测炎症及凋亡相关蛋白表达水平；DCFH-DA染色法、化学比色法检测细胞内活性氧（ROS）及超氧化物歧化酶（SOD）活力、丙二醛（MDA）含量。结果PQ能明显抑制HK-2细胞增殖，L6H21可明显缓解PQ的抑制作用。与阴性对照组比较，PQ组HK-2细胞凋亡明显升高，Caspase-3、Caspase-9、Bax水平升高，Bcl-2表达下降，差异有统计学意义（P〈0．05）；与PQ组比较，PQ＋L6H21组Caspase-3、Caspase-9、Bax水平下降，Bcl．2表达升高，差异有统计学意义（P〈0．05）。与阴性对照组比较，PQ组NF．KB、TNF-α、IL-6表达明显升高，差异有统计学意义（P〈0.05），与PQ组比较，PQ＋L6H21组NF-KB、TNF-α、IL-6水平下降，差异有统计学意义（P〈0．05）。与阴性对照组比较，PQ组HK．2细胞内ROS及MDA水平明显升高，SOD活力降低，差异有统计学意义（P〈0．05），PQ＋L6H21组ROS、MDA水平降低，SOD活力升高，差异有统计学意义（P〈0．05）。结论L6H21能够明显减轻PQ诱导的HK-2损伤和凋亡，其机制可能与缓减PQ诱导的炎症反应和氧化损伤有关。
Objective To investigate the effects of mono-carbonyl analogues of curcumin （L6H21） on paraquat （PQ）-induced injury in HK-2 cell line and explore its underlying mechanisms. Methods Cultured HK-2 cells were challenged by PQ with or without L6H21 treatment. Cell viability and apoptosis were determined by CCK-8 assay and flow cytometry, respectively. Gene expressions and protein levels of apoptotie and inflammatory factors were assessed by RT-PCR, ELISA, and western blot. Intracellular ROS production was detected by DCFH- DA staining. Superoxide dismutase （SOD） and malondialdehyde （MDA） were examined by chemical colorimetry. Results 1） PQ challenge significantly inhibited HK-2 cells proliferation, which was prevented by L6H21 administration. PQ dramatically induced HK-2 apoptosis evidenced by increasing expressions of caspase-9, caspase-3 and Bax, while decreasing Bcl-2 level. However, PQ induced these apoptotic effects in HK-2 cells were reversed by L6H21. Similarly, PQ exposure obviously enhanced activity of NF-KB and levels of cytokines （TNF- α ,IL-6） in HK-2 cells, which was inhibited by L6H21. Furthermore, administration of L6H21 inhibited PQ induced ROS and MDA production, and promoted SOD level in HK-2 cells. Conclusion L6H21 administration inhibits PQ-indueed apoptosis in HK-2 cells possibly by reducing inflammation and oxidative damage.
出处
《中华劳动卫生职业病杂志》
CAS
CSCD
2017年第9期641-647,共7页
Chinese Journal of Industrial Hygiene and Occupational Diseases
基金
浙江省中医药科技计划项目（2013ZA088）
药卫生平台骨干人才计划（2016RCA021）
浙江省中医药重点学科计划（2012-XK-A28）
关键词
姜黄素
单羰基姜黄素类似物
百草枯
上皮细胞
Curcumin
Mono-carbonyl analogues of curcumin
Paraquat
Epithelial cell
