阿米洛利对PC12细胞的保护作用及其对LAMP2a表达影响  

Amiloride protects PC12 cells from acid-induced injury via down-regulation the expression of LAMP2a

作　　者：赵长振[1,2] 徐耑[3] 张润娉[3] 孔岩[3] 

机构地区：[1]苏州高新区狮山街道社区卫生服务中心,江苏苏州215011 [2]苏州大学 [3]苏州大学附属第一医院神经内科,江苏苏州215006

出　　处：《基础医学与临床》2011年第10期1124-1128,共5页Basic and Clinical Medicine

摘　　要：目的研究阿米洛利对PC12细胞的保护作用,及其对LAMP2a表达水平的影响。方法 pH 6.0的酸性培养液诱导PC12细胞损伤,同时给予阿米洛利进行干预,四甲基偶氮唑盐(MTT)、乳酸脱氢酶(LDH)和流式细胞技术检测细胞存活率、损伤程度及凋亡率,Western blot检测LAMP2a表达水平。结果酸性培养液处理24 h,细胞存活率降低至(67.2±5.1)%,上清液中的LDH增多至(407.9±11.1)U/L,凋亡率升高至(27.8±2.6)%。阿米洛利(100μmol/L)提高PC12细胞的存活率至(84.9±2.2)%,LDH降低至(314.1±5.2)U/L,细胞凋亡率降低至(17.8±1.3)%,与pH 6.0组相比(P<0.001)。酸处理导致LAMP2a表达升高,而阿米洛利能够显著抑制LAMP2a的表达(P<0.05)。结论阿米洛利通过下调LAMP2a的表达发挥神经保护作用。Objective To study the neuroprotective effects of amiloride（Ami）,non-selective blocker of acid-sensing ion channels,in the cell death and apoptosis induced by extracellular acid in PC12 cells,and detect the expression of LAMP2a.Methods The cell viability following acid exposure（pH 6.0） was analyzed with 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assay.Cell injury was examined by a quantitative measurement of lactate dehydrogenase（LDH） released from cytosol to culture medium.Double staining of treated cells with Annexin V and PI was assayed with flow cytometry to determine the apoptotic rate.Western blot was used to examine the expression levels of receptor lysosome-associated membrane protein 2a（LAMP2a）.Results Acid exposure significantly decreased the cell viability（67.2±5.1）%,increased the activity of LDH（407.9±11.1）U/L which released to culture medium and raised apoptosis rate（27.8±2.6）%.Western bolt analysis shown that acidic exposure led to compensatory induction of LAMP2a protein signaficantly（P0.01）.Co-incub-ation with Ami（100 μmol/L） significantly increased the cell viability（84.9±2.2）%,inhibited the release of LDH（314.1±5.2）U/L,reduced the apoptosis rate（17.8±1.3）%,and inhibited the overexpression of LAMP2a（P0.05）.Conclusions Ami protects PC12 cells against acid-induced injury via down-regulation the expression of LAMP2a.

关 键 词：阿米洛利 LAMP2a 分子伴侣自噬 PC12细胞 帕金森病 

分 类 号：R965[医药卫生—药理学]