CAJ | 학술논문

目的：体外培养大鼠 INS-1细胞,观察间歇低氧条件直接作用于细胞引起的改变。方法常氧组细胞(CON)不进行间歇低氧暴露,IH 组细胞暴露间歇低氧,收集暴露(1、2、3 d)的培养上清及细胞,检测氧化应激指标超氧化物歧化酶(SOD)、丙二醛(MDA)及 JNK1 mRNA、JNK1/2蛋白表达水平。给予 SP600125阻断 JNK 蛋白活性,观察下游基因 PDX-1 mRNA 及蛋白变化。结果随着间歇低氧暴露时间的延长,IH 组细胞培养上清中 MDA 含量呈增高趋势,SOD 活力呈降低趋势。 IH 组细胞表达 JNK1 mRNA 较相应时间 CON 组明显增高,其表达量进行性下降。磷酸化 JNK1于 IH 3 d 组呈现出高表达。给予 SP600125可引起 PDX-1蛋白明显增高。结论间歇低氧可以直接引起INS-1细胞氧化损伤,随暴露时间的延长而加重,可能是通过诱导 JNK1转录水平及蛋白活化而实现的。
목적：체외배양대서 INS-1세포,관찰간헐저양조건직접작용우세포인기적개변。방법상양조세포(CON)불진행간헐저양폭로,IH 조세포폭로간헐저양,수집폭로(1、2、3 d)적배양상청급세포,검측양화응격지표초양화물기화매(SOD)、병이철(MDA)급 JNK1 mRNA、JNK1/2단백표체수평。급여 SP600125조단 JNK 단백활성,관찰하유기인 PDX-1 mRNA 급단백변화。결과수착간헐저양폭로시간적연장,IH 조세포배양상청중 MDA 함량정증고추세,SOD 활력정강저추세。 IH 조세포표체 JNK1 mRNA 교상응시간 CON 조명현증고,기표체량진행성하강。린산화 JNK1우 IH 3 d 조정현출고표체。급여 SP600125가인기 PDX-1단백명현증고。결론간헐저양가이직접인기INS-1세포양화손상,수폭로시간적연장이가중,가능시통과유도 JNK1전록수평급단백활화이실현적。
Objective To observe the direct effect on INS-1 cells of rats cultured in vitro under intermittent hy-poxia. Methods Cells in common oxygen group (CON group) was not exposed under intermittent hypoxia, while cells in intermittent hypoxia group (IH group) were exposed under intermittent hypoxia. Cultured supernate and cells were col-lected at 1st d, 2nd d and 3rd d after exposure and the levels of superoxide dismutase (SOD), malondialdehyde (MDA) and expressions of JNK1 mRNA and JNK1 / 2 in the two groups were detected. SP600125 was used to bolck the activity of JNK, and changes of downstream gene PDX-1 mRNA and protein were also observed. Results The MDA level at super-nate was significantly higher and SOD activity was significantly decreased in cultured supernate of IH group with prolonged exposure time under intermittent hypoxia. JNK1 mRNA expression was significantly upregulated in IH group compared with that in CON group at the same time, and expression value was progressively decreased. The high phospho-JNK1 ex-pression was found 3th d after exposure. The expression of PDX-1 was significantly increased after using SP600125. Con-clusion Intermittent hypoxia may directly induce oxidative damage effect on INS-1 cells, and the damage becomes seri-ous with time prolonging, and its damage may be done by inducing transcriptional level and protein activation.