缺氧及复氧对去整合蛋白和金属蛋白水解酶10和淀粉样β分泌酶mRNA水平的影响

Effect of hypoxia and reoxygenation on the expression of a disintegrin and metalloprotease10,β-amyloid precursor protein cleaving enzyme mRNA in SH-SY5Y cells

目的研究缺氧及复氧损伤对去整合蛋白和金属蛋白水解酶10(ADAM10)和淀粉样β分泌酶(BACE1)mRNA的影响。方法研究分为缺氧12 h组,缺氧24 h组及各自对照组。缺氧组在缺氧条件下(95%N2,5%CO2)分别培养SH-SY5Y细胞12 h和24 h后复氧,分别于0、12、24 h利用逆转录-聚合酶链反应(RT-PCR)方法检测ADAM10、BACE1 mRNA水平;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12 h后复氧各时间点ADAM10 mRNA表达下调(分别较对照组下降19.8%、41.4%、64.6%;P=0.005、0.038、0.001),缺氧培养24 h后复氧各时间点ADAM10 mRNA表达下调更明显(分别较对照组下降30.1%、75.9%、86.5%;P=0.009、0.005、0.043);BACE1 mRNA在缺氧24 h后复氧12 h和24 h表达上调(分别较对照组上调31.5%和35.1%;P=0.028、0.005)。结论缺氧及复氧可以降低ADAM10转录水平和增加BACE1转录水平。

Objective To study the effects of hypoxia and reoxygenation on expression of a disintegrin and metalloprotease10 （ADAM10）, β-amyloid precursor protein cleaving enzyme （BACE1） mRNA in SH-SY5Y cells. Methods SH-SY5Y cells were grouped into hypoxic and control groups. The hypoxic cells were incubated in hypoxic condition （95%N2, 5%CO2） for 12 hours and 24 hours, and then cells were reoxygenated for 0 hours, 12 hours and 24 hours. The expression of ADAM10, BACE1 mRNA were tested respectively at different time points after reoxygenation by RT-PCR. Control groups were incubated in normal conditions, seeded and treated at the same time with the hypoxic cells. Results The expression of ADAM10 mRNA was down-regulated by 19.8%, 41.4% and 64.6% （P = 0.005, 0.038, 0.001） at different time after reoxygenation with 12 hours hypoxia and down-regulated by 30.1%, 75.9% and 86.5% （P = 0.009, 0.005, 0.043） after reoxygenation with 24 hours hypoxia. The expression of BACE1 mRNA was up-regulated by 31.5% and 35.1% （P = 0.028, 0.005） only at 12 hours and 24 hours points after reoxygenation with 24 hours hypoxia. Conclusion Hypoxia and reoxygenation might alter the expression of ADAM10 and BACE1, which demonstrates that the vascular factors should make the amyloid precursor protein easy to be processed by β-secrease pathway, thus to involve the pathology of Alzheimer＇s disease.