CAJ | 학술논문

目的探讨沙利度胺(THD)对百草枯诱导的小鼠急性肺损伤的干预及其机制.方法清洁级雄性ICR小鼠随机分成空白对照组(30只)、沙利度胺对照组(30只)、百草枯中毒组(30只)和不同剂量的沙利度胺干预组(90只),分别于不同处理1、3、7天麻醉处死小鼠,取肺组织.光学显微镜下观察各组小鼠肺组织病理表现;TUNEL染色法观察肺组织细胞凋亡;实时反转录聚合酶链式反应(RT-PCR)测定肺组织核因子Nrf2的mRNA表达;提取肺组织细胞核蛋白,蛋白免疫印迹法检测胞核内Nrf2蛋白量.化学比色法测量小鼠肺组织丙二醛(MDA)含量、超岐化物氧化酶(SOD)活力、谷胱甘肽(GSH)含量.结果与空白对照组比较,百草枯中毒组肺组织Nrf2 mRNA和核内Nrf2蛋白量表达量升高,MDA及SOD活力升高,GSH含量降低,差异有统计学意义(P＜0.05).与百草枯中毒组比较,沙利度胺干预组肺组织Nrf2 mRNA和核内Nrf2蛋白表达量升高,MDA含量降低,SOD活力和GSH含量升高,差异有统计学意义(P＜0.05).肺组织病理学显示,百草枯中毒组在染毒后3d时肺损害明显,沙利度胺干预组肺损害减轻.结论沙利度胺能明显减轻急性百草枯中毒小鼠的肺组织损伤,其机制可能与影响Nrf2信号通路,平衡氧化与抗氧化失衡和抑制细胞凋亡有关.
목적 탐토사리도알(THD)대백초고유도적소서급성폐손상적간예급기궤제.방법 청길급웅성ICR소서수궤분성공백대조조(30지)、사리도알대조조(30지)、백초고중독조(30지)화불동제량적사리도알간예조(90지),분별우불동처리1、3、7천마취처사소서,취폐조직.광학현미경하관찰각조소서폐조직병리표현;TUNEL염색법관찰폐조직세포조망;실시반전록취합매련식반응(RT-PCR)측정폐조직핵인자Nrf2적mRNA표체;제취폐조직세포핵단백,단백면역인적법검측포핵내Nrf2단백량.화학비색법측량소서폐조직병이철(MDA)함량、초기화물양화매(SOD)활력、곡광감태(GSH)함량.결과 여공백대조조비교,백초고중독조폐조직Nrf2 mRNA화핵내Nrf2단백량표체량승고,MDA급SOD활력승고,GSH함량강저,차이유통계학의의(P＜0.05).여백초고중독조비교,사리도알간예조폐조직Nrf2 mRNA화핵내Nrf2단백표체량승고,MDA함량강저,SOD활력화GSH함량승고,차이유통계학의의(P＜0.05).폐조직병이학현시,백초고중독조재염독후3d시폐손해명현,사리도알간예조폐손해감경.결론 사리도알능명현감경급성백초고중독소서적폐조직손상,기궤제가능여영향Nrf2신호통로,평형양화여항양화실형화억제세포조망유관.
Objective To investigate the effects of thalidomide in a mouse model of paraquat-induced acute lung injury and the mechanisms underlying the properties of thalidomide.Methods Male ICR mice were randomly allocated into four groups:nomal control group(n=30),thalidomide control group (n=30),paraquat poisioning group (n=30) and thalidomide treatment group (n=90).Mice were sacrificed at 1d,3d and 7d after paraquat poisioning.The level of (MDA)malondialdehyde,Superoxidedi-smutase (SOD) and glutathione (GSH) in the lung tissue were measuerd by chemical colorimetry.The expression of Nrf2 mRNA was determined by RT-PCR; Nuclear protein Nrf2 was abserved by Western-blotting; Pathological changes of lung tissue were observed under light microscope by HE stain; the lung apoptosis cells were detected by TUNEL.Results The levels of MDA,SOD and the expressions Nrf2 mRNA and protein Nrf2 in lung tissue were all markedly increased in mice of paraquat poisioning group than those in nomal group at 1 d,3 d,7 d.In contrast,the levels of GSH were decreaseel (P＜0.05).Compared with paraquat poisioning group,the pulmonary SOD,Nrf2 mRNA and protein were increased and the lung wet dry ratio were all significantly decreased in mice of THD treatment group at 1 d,3 d,7 d (P＜0.05).THD alleviated the pulmonary damage in the lightmicroscope at 3d after paraquat poisioning.The apoptosis index was markedly decreased in THD treatment groups comparing to paraquat piosioning group (P＜0.05).Conclusions Lipid peroxide damage was one of the mechanisms of paraquat poisioning,thalidomide could attenuate paraquat-induced acute lung injury and its mechanism may be activating the Nrf2-ARE signaling pathway to protect mouse from Lipid peroxide damage.