CAJ | 학술논문

目的研究姜黄素(curcumin)对百草枯(paraquat,PQ)诱导的Ⅱ型肺泡细胞氧化损伤的保护作用及机制.方法常规培养A549细胞,分空白对照组、姜黄素对照组、PQ组及姜黄素+PQ组,处理24h,MTT法检测细胞存活率、RT-PCR及Western blot法检测A549细胞NFE2L2表达、ELISA法检测细胞内HO-1、NQO-1及上清液超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活力及丙二醛(MDA)水平.并以siRNA技术低表达Nrf2,观察其对姜黄素拮抗PQ诱导A549细胞氧化损伤指标的影响.结果 0.1 mmol/L PQ即能明显抑制A549细胞生长,并呈剂量依赖性;而160 μmol/L以下姜黄素对A549细胞生长无明显抑制作用.800 μmol/L PQ组A549细胞SOD活力明显降低,CAT活力及MDA水平均明显升高,与空白对照组比较,差异有统计学意义(P＜0.05,P＜0.01).80 μmol/L姜黄素+PQ组,SOD及CAT活力较PQ组明显升高,MDA水平明显降低,差异有统计学意义(P＜0.01).与对照组比较,PQ染毒组NFE2L2及其下游因子HO-1、NQO-1表达均明显升高,差异有统计学意义(P＜0.01);姜黄素+PQ组NFE2L2及HO-1、NQO-1表达水平较PQ组升高,差异均有统计学意义(P＜0.01).与姜黄素+PQ组比较,姜黄素+PQ+NFE2L2siRNA组SOD、CAT活力均明显下降,MDA水平明显升高,差异有统计学意义(P＜0.01).结论小剂量姜黄素在体外能明显拮抗PQ诱导的A549细胞氧化损伤,这种保护作用依赖于Nrf2-ARE通路的活化.
목적 연구강황소(curcumin)대백초고(paraquat,PQ)유도적Ⅱ형폐포세포양화손상적보호작용급궤제.방법 상규배양A549세포,분공백대조조、강황소대조조、PQ조급강황소+PQ조,처리24h,MTT법검측세포존활솔、RT-PCR급Western blot법검측A549세포NFE2L2표체、ELISA법검측세포내HO-1、NQO-1급상청액초양화물기화매(SOD)、과양화경매(CAT)활력급병이철(MDA)수평.병이siRNA기술저표체Nrf2,관찰기대강황소길항PQ유도A549세포양화손상지표적영향.결과 0.1 mmol/L PQ즉능명현억제A549세포생장,병정제량의뢰성;이160 μmol/L이하강황소대A549세포생장무명현억제작용.800 μmol/L PQ조A549세포SOD활력명현강저,CAT활력급MDA수평균명현승고,여공백대조조비교,차이유통계학의의(P＜0.05,P＜0.01).80 μmol/L강황소+PQ조,SOD급CAT활력교PQ조명현승고,MDA수평명현강저,차이유통계학의의(P＜0.01).여대조조비교,PQ염독조NFE2L2급기하유인자HO-1、NQO-1표체균명현승고,차이유통계학의의(P＜0.01);강황소+PQ조NFE2L2급HO-1、NQO-1표체수평교PQ조승고,차이균유통계학의의(P＜0.01).여강황소+PQ조비교,강황소+PQ+NFE2L2siRNA조SOD、CAT활력균명현하강,MDA수평명현승고,차이유통계학의의(P＜0.01).결론 소제량강황소재체외능명현길항PQ유도적A549세포양화손상,저충보호작용의뢰우Nrf2-ARE통로적활화.
Objective To investigate the protective effect ofcurcumin (CU) on type Ⅱ alveolar epithelial cells (A549 cells) during paraquat (PQ)-induced oxidative damage and its underlying mechanism.Methods Routinely cultured A549 cells were divided into blank control group,CU control group,PQ group,and PQ+Cu group to receive respective treatments for 24 h.Cell viability was determined by MTT assay.The NFE2L2 expression in A549 cells was measured by RT-PCR and Western blot.The activities of the heme oxygenase-l (HO-1) and NAD(P)H:quinone oxidoreductase 1 (NQO-1) in cells and the superoxide dismutase (SOD) anid catalase (CAT) in supernatant,as well as malondialdehyde (MDA) content,were measured by enzyme-linked immunosorbent assay.After siRNA depletion of Nrf2,the protective effect of CU on A549 cells during PQinduced oxidative damage was evaluated.Results PQ,even at a dose of 0.1 mmol/L,could significantly suppress the viability of A549 cells in a dose-dependent manner.CU showed no significant inhibitory effect on the viability of A549 cells when given at a dose below 160 μmol/L.Compared with the blank control group,the PQ group had significantly decreased SOD activity and significantly increased CAT activity and MDA content after 24-h exposure to 800 μmol/L PQ (P＜0.05 or P＜0.01).Thanks to pretreatment with 80 μmol/L CU,the PQ+CU group had significantly increased SOD and CAT activities and significantly decreased MDA content compared with the PQ group (P＜0.01).Compared with the blank control group,the PQ group had significantly increased expression of NFE2L2 and its downstream factors HO-1 and NQO-1 (P＜0.01),while the PQ+CU group had significantly higher expression of NFE2L2,HO-1,and NQO-1 than the PQ group (P＜0.01).Compared with the PQ+ CU group,the CU+PQ+NFE2L2siRNA group had significantly decreased SOD and CAT activities and significantly increased MDA content (P＜0.01).Conclusion Low-dose CU significantly reduces the PQ-induced oxidative damage in A549 cells in vitro by activation of the Nrf2-ARE pathway.