细胞内酸化对自噬的影响  

Influence of intracellular acidification on autophagy

作　　者：肖楠[1] 方伯言[2] 

机构地区：[1]辽宁医学院研究生院,辽宁锦州121001 [2]辽宁医学院附属第一医院

出　　处：《山东医药》2012年第9期17-19,共3页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81100962);辽宁省博士科研启动;自然科学基金资助项目(20082179);辽宁省教育厅高等学校科研项目(20060524)

摘　　要：目的探讨细胞内酸化对自噬的影响。方法利用pAd-miR-NHE-1病毒悬液转染人源性神经母细胞瘤细胞株,制备NHE-1基因敲低细胞模型。在透射电镜下观察阴性对照组及腺病毒转组转染成功后0.5、2、4、8 h自噬体的形成、线粒体形态及溶酶体数目的变化;Western blotting法检测自噬标记物LC3Ⅱ的表达。结果透射电镜下,阴性对照组线粒体形态及溶酶体数目正常,腺病毒转染成功后0.5～4 h可见线粒体肿胀、变形,溶酶体数目增多,自噬囊泡增多,自噬小体形成。8 h可见凋亡小体出现,线粒体数目减少,未见自噬小体,可见少量溶酶体。Western blotting显示,阴性对照组LC3Ⅱ表达较低,腺病毒转染成功后0.5 h LC3Ⅱ表达升高(P<0.01),4 h达高峰(P<0.01),8 h表达降低,且低于阴性对照组(P<0.01)。结论细胞内酸化可以激活自噬的发生。Objective To explore the influence of intracellular acidification on autophagy. Methods Using the pAd- miR-NHE1 virus levitation liquid to infect SH-SY5Y cells, for the preparation of cell model of low NHE-1 gene, observing the formation of autophagy, changes of mitoehondrial form and lysosome number of negative control group, the group after the succesful transfection 0.5, 2, 4, 8 h of adenovirus levitation liquid by transmission electron microscopy, testing the ex- pression of autophagy marker-LC3 by Western blotting. Results Through the transmission electron microscopy, the normal mitochondrial form and lysosome number were shown in negative control group. After the succesful transfeetion 0.5-8 h of adenovirus levitation liquid, swelling mitochondria, the lysosome populations, autophagy vesicles increased, the autophagy body formed. After 8 hours, apoptotie body appeared, mitochondria numbers decreased, no autophagy body, a bit lyso- somes. There was a low LC-3 expression in negative control group, after the succesful transfection 0.5 h, the expression of LC-3 rose （P 〈0.01 ）, then reached the peak in 4 h （P 〈0.01 ）. In 8 h, the expression of LC-3 was lowest, it was lower than that in negative control group （P 〈 0.01 ）. Conclusions Cells acidification could activate the happening of autoph- agy.

关 键 词：细胞内酸化 自噬 LC3 

分 类 号：Q78[生物学—分子生物学]