缺氧及复氧对淀粉样前体蛋白、去整合蛋白和金属蛋白水解酶10和淀粉样β分泌酶蛋白表达的影响

Effect of Hypoxia and Reoxygenation on the protein expression of amyloid precursor protein,a disintegrin and metalloprotease10 and β-amyloid precursor protein cleaving enzyme

目的研究缺氧及复氧损伤对淀粉样前体蛋白(APP)、去整合蛋白和金属蛋白水解酶10(ADAM 10)和淀粉样β分泌酶(BACE 1)蛋白表达的影响。方法分为缺氧组和对照组。缺氧组在缺氧条件下(95%N2,5%CO2)培养SH-SY 5Y细胞24h后复氧,分别于0h、12h、24h利用W estern-b lot方法检测APP、ADAM 10和BACE 1蛋白表达;对照组在正常情况下培养,与缺氧组同一时间进行接种和处理。结果缺氧培养SH-SY 5Y细胞24h后复氧各时间点APP蛋白表达与对照组相比差别无统计学意义(P=0.818,0.728和0.674);ADAM 10蛋白表达分别较对照组下降25.5%,67.3%和75.9%(P=0.001,0.001,0.000);BACE 1蛋白表达分别较对照组增加22%,42%和51.5%(P=0.021,0.000和P=0.000)。结论缺氧及复氧可以降低ADAM 10蛋白表达和增加BACE 1蛋白表达。提示血管因素可能使APP更易通过产生淀粉样肽(Aβ)途径裂解,而参与SAD的病理过程。

Objective To study the effect of hypoxia and reoxygenation on the protein expression of amyloid precursor protein （APP）, a disintegrin and metalloprotease 10 （ADAM 10 ）and β-amyloid precursor protein cleaving enzyme （BACE1）. Methods SH-SYSY cells were grouped into hypoxic and control groups. The hypoxic cells were incubated in hypoxic condition（95%N2,5%CO2）for 24h,then cells were reoxygenated for 0h,12h and 24h. Western blotting was performed to examine the protein expression of APP,ADAM 10 and BACE1 at different time points after reoxygenation. Control groups were incubated in normal conditions,seeded and treated at the same time with the hypoxic cells. Results Reoxygenation for different time after 24h-hypoxia did not alter the protein expression of APP （P = 0. 818,0. 728,0. 674）, made the protein expression of ADAM10 decrease by 25. 5%, 67.3%,75.9%（P=0. 001,0. 001,0. 000）and the protein expression of BACE1 increased 22%,42% ,51.5% （P= 0. 021,0. 000 0. 000）. Conclusion Hypoxia and reoxygenation might alter the protein expression of ADAM10 and BACE1 ,which demonstrates that the vascular factors should make the amyloid precusor protein easy to be processed by β-secretase pathway.and involve into the pathology of sporadic Alzheimer＇s disease.