IκBα突变型基因对缺氧缺糖损伤永生化神经前体细胞的保护作用　.doc

I K B a突变型基因对缺氧缺糖损伤永生化神经前体细胞的保护作用 ：祝畅刘志恒张传汉桂伶俐姚文龙 】目的：检测在缺氧缺糖环境中，Ik Ba突变 型基因对永生化神经前体细胞株（INPCs） NF?kB的活性、 细胞存活及细胞损伤的影响.方法:在建立转染及转染/I k B a MINPCs的基础上，用MTT法检测缺氧缺糖处理3，6和 9h后两细胞株的存活率，用焚光素酶报告基因检测两细胞 株缺氧缺糖6h前后NF? k B的活性，并测定缺氧缺糖6h处 理后培养液中乳酸脱氢酶的含量，用Hoechs t33 342染色 观察细胞核的形态.结果：转染/I k B a M的I NPCs中N F? k B的活性均低于转染的I NPCs,缺氧缺糖6 h或9h时， 前者存活率高于后者，且6h时前者上清中的乳酸脱氢酶释 放量较少，缺氧缺糖可使两细胞株的部分细胞核呈现凋亡 改变.结论：I KBa突变型基因可降低INPCs中N F?kB的 活性，提高INPCs在缺氧缺糖处理后的存活率，减轻细胞损 伤. 关键词】干细胞；NF?kB抑制因子a;细胞缺氧 Abstra ct] AIM:Tod e termineth ee ffectofm uta tedl KBa gene onimmo rtali zedne uralpr ogen itorcel 1st rains(IN PC s)inoxyge n ?glucosede privation. M ETHODS:Afte rtheestabl i shmentofm ut atedlk B a gen etransf ecte dINPCs andbl ankpl asmidt rans fectedl NPC s,thecel Is wereexpos e dtoanoxic/ hypoglycemi cconditionf or3,6or9hr e spectivel y. MTTstain ing wasused tode tectth esurv ivalr ateoft hetw ocellst rai ns. Lucif er aseassays y stemwasuse dtodetectth eactivityof NF? k Bintra n sfectedor /I k B a Mtran sfe ctedINP Cs. L actate dehyd rogen aseinc ultu remediu mwa smeasure da fterthece 1 lswereexpo sedtoanoxic /hypoglycem icconditio n for6h. The mo rphologi cal changeo ftwo cellst rains wasob served byHo echst33 342 staining . R ESULTS:Th e activityof NF? k Bwasdow n?regulated in/Ik B a Mtr a nsfectedl NP Csbefore ora fteroxy gen?glucos edepr ivati on. The cell surviva Ira teof/I k B a M transfect e dINPCswash igherthanth atoftransfe ctedINPCsa f terexpose dt oanoxic/hyp oglycem icco nditio nfor6 hor9h , butth erew asnodif fer enceince 11 survivalr a teat3h. Aft erthecellsw ereexposedt oanoxic/hy p oglycemic co nditionf or6 h, lacta tede hydrog enase conte ntincu ltur emedium was lowerin/ I k B a Mtransf e ctedINPCs. Hoechst3334 2stainingre vealedthat c ellswerea po ptoticaf ter oxygen?gluc osedep rivat ion.C ONCLUS ION:Bydown?reg ulatingt he activityo f NF? k B, muta tedl k B a gene improvesthe cellsurviv a lrateofIN PC sandless ens thecell dama gecaus edbyo xygen ?gluco sede privati on. [Keywo