Evaluation of Enzyme Induction in Alcohol Metabolism by using RT-PCR.

众所周知，东方冲红犬纯合子携带aldh2 *2，即使摄入小剂量的乙醇也会表现出烦躁不安的症状。然而，杂合子受试者（ALDH2*2/*1）可以少量饮酒，几小时后再多喝一点，第二次挑战时烦躁症状有所减轻。本研究的主要目的是确定乙醇摄入前后ALDH2 mRNA水平是否有差异。采用RT-PCR技术，建立了测定外周血ALDH2 mRNA水平的方法。从健康日本志愿者外周血淋巴细胞中提取无dna总RNA。将纯化的RNA逆转录为cDNA。利用引物（通过外显子11和12-13设计）进行PCR扩增。PCR DNA产物

It is well known that the Oriental flushers, homozygotically possessingALDH2*2, show dysphoric symptoms after intake of even small doses of ethanol. Heterozygotic subjects (ALDH2*2/*1) can, however, drink a small amount and then after a few hours drink a little more alcohol, with the dysphoric symptoms diminish somewhat to the second challenge. The main purpose of this study was to determine whether the levels of mRNA for ALDH2 are different before and after ethanol intake.A method for determining mRNA levels for ALDH2 in peripheral blood was developed using an RT-PCR technique. DNA-free total RNA was extracted from peripheral lymphocytes from healthy Japanese volunteers. Purified RNA was reverse transcribed into cDNA.The cDNA was amplified with PCR using the primers, designed through exon 11 and 12-13. PCR DNA products were