CAJ | 학술논문

目的观察不同剂量米非司酮(RU486)诱导Nrf2基因表达对百草枯(PQ)致A549细胞毒性影响.方法 Ad-RUNff2感染的A549细胞,在给予10-10、10-9、10-8、10-7 mol/L的RU486诱导表达6h后,加入浓度为10-3 mol/L的PQ培养48 h,用实时定量聚合酶链式反应(real-time PCR)法和凝胶迁移率试验( EMSA)检测不同浓度RU486诱导的Nrf2表达情况；Real-timePCR、ELISA检测不同浓度Nrf2对PQ致A549细胞损伤炎症因子白细胞介素(IL)-6、IL-10、肿瘤坏死因子(TNF)-α,凋亡因子半胱氨酸天冬氨酸蛋白酶( Caspase )-3、Caspase-9、细胞色素c(Cyt C)基因相对表达量和蛋白含量的影响.用化学比色法检测氧化因子过氧化氢酶(CAT)、丙二醛(MDA)蛋白含量.结果 Nrf2基因相对表达量及蛋白表达均随RU486浓度增高而增高,其中以RU486浓度为10-7 mol/L时最高,与PQ中毒组及空白对照组的差异有统计学意义(P＜0.01,P＜0.05).随RU486浓度降低,IL-6、TNF-α的基因相对表达量及蛋白含量升高,在RU486 10-7 mol/L时最低,与PQ中毒组及空白对照组的差异有统计学意义(P＜0.01,P＜0.05)；IL-10的变化则相反.随RU486浓度降低,Caspase-3、Caspase-9、Cyt C基因相对表达量升高,10-7 mol/L时最低,与PQ中毒组及空白对照组的差异有统计学意义(P＜0.01,P＜0.05).随着RU486浓度的增高,CAT 蛋白含量增高,RU486浓度为10-7 mol/L时含量最高,与PQ中毒组及空白对照组的差异有统计学意义(P＜0.05),MDA含量的变化则相反.结论 RU486诱导Nrf2基因表达能促进A549细胞的氧化-抗氧化系统平衡,并抑制炎症因子、凋亡因子,对PQ致A549细胞损伤有保护作用.
목적 관찰불동제량미비사동(RU486)유도Nrf2기인표체대백초고(PQ)치A549세포독성영향.방법 Ad-RUNff2감염적A549세포,재급여10-10、10-9、10-8、10-7 mol/L적RU486유도표체6h후,가입농도위10-3 mol/L적PQ배양48 h,용실시정량취합매련식반응(real-time PCR)법화응효천이솔시험( EMSA)검측불동농도RU486유도적Nrf2표체정황；Real-timePCR、ELISA검측불동농도Nrf2대PQ치A549세포손상염증인자백세포개소(IL)-6、IL-10、종류배사인자(TNF)-α,조망인자반광안산천동안산단백매( Caspase )-3、Caspase-9、세포색소c(Cyt C)기인상대표체량화단백함량적영향.용화학비색법검측양화인자과양화경매(CAT)、병이철(MDA)단백함량.결과 Nrf2기인상대표체량급단백표체균수RU486농도증고이증고,기중이RU486농도위10-7 mol/L시최고,여PQ중독조급공백대조조적차이유통계학의의(P＜0.01,P＜0.05).수RU486농도강저,IL-6、TNF-α적기인상대표체량급단백함량승고,재RU486 10-7 mol/L시최저,여PQ중독조급공백대조조적차이유통계학의의(P＜0.01,P＜0.05)；IL-10적변화칙상반.수RU486농도강저,Caspase-3、Caspase-9、Cyt C기인상대표체량승고,10-7 mol/L시최저,여PQ중독조급공백대조조적차이유통계학의의(P＜0.01,P＜0.05).수착RU486농도적증고,CAT 단백함량증고,RU486농도위10-7 mol/L시함량최고,여PQ중독조급공백대조조적차이유통계학의의(P＜0.05),MDA함량적변화칙상반.결론 RU486유도Nrf2기인표체능촉진A549세포적양화-항양화계통평형,병억제염증인자、조망인자,대PQ치A549세포손상유보호작용.
Objective To observe the effects of Nrf2 gene expression induced by RU486 at different doses on A549 cell damage induced by paraquat (PQ).Methods After A549 cells transfected with AdRUNrf2 were treated by RU486 at the doses of 10-10,10-9,10-8 and 10q mol / L for 6 h,A549 cell cultures were exposed to 10-3 mol/L of PQ for 48 h.Then qRT-PCR and EMSA assays were used to detect the expression of Nrf2 gene,and qRT-PCR and ELISA assays were utilized to measure the effects of Nrf2 gene on the expression of the inflammatory cytokines IL-6,IL-10 and TNF-α,apoptotic factors Caspase-3,Caspase-9 and Cytochrome C.The oxidation factors (CAT and MDA protein contents) were observed by Chemical Colorimetric Analysis.Results Nrf2 gene relative expression and protein contents increased with RU486 concentrations,and the above expression was the highest when the concentration of RU486 was 10-7 mol/L,which was significantly higher than those in control and PQ exposure groups (P＜0.01 or P＜0.05).The relative gene expression and protein expression of IL-6 and TNF-α enhanced with the reduced concentrations of RU486,which were the lowest when RU486 concentration was 10-7 mol/L,as compared with control and PQ exposure groups (P＜0.01 or P＜0.05),while the change of IL-10 content was the opposite.The relative expression of Caspase3,Caspase9 and Cytochrome C genes also increased with the reduced concentrations of RU486,which were the lowest when RU486 concentration was 10-7 mol/L,as compared with control and PQ exposure groups (P＜0.01 or P＜0.05).The content of CAT enhanced with RU486 concentration,which was the highest when RU486 concentration was 10-7 mol/L,as compared with control and PQ exposure groups (P＜0.05).But the change of MDA content was the contrary.Conclusion Nrf2 expression induced by RU486 can promote the balance of oxidation-antioxidation system in A549 cells and inhibit the inflammation and apoptosis factors,which has a protective effect on A549 cell injury induced by PQ.