Expression of alpha-1-microglobulin mRNA change after oxalate challenge in renal epithelium cell

• Main Authors: Jeng-Yu Tsai, 蔡政諭
• Other Authors: David, Chao
• Format: Others
• Language: zh-TW
• Published: 2002
• Online Access: http://ndltd.ncl.edu.tw/handle/13173176619415806067

碩士 === 國立中山大學 === 生物科學系研究所 === 90 === Alpha-1-microglobulin is co-synthesized with bikunin on the AMBP gene (□-1m-bikunin precursor gene). The major synthesis site of this protein is liver. It is excreted into urine by pass through glomerulus. Both alpha-1-microglobulin and bikunin are regarded as inhibitors of urolithiasis. This investigation is attempted to evaluate if mRNA in renal epithelium cell may be up-regulated by oxalate and oxalate crystal stimulation. LLC-PK1 cells were challenged with oxalate and calcium oxalate crystal. The messenger RNA was collected and reverse transcripted into cDNA. PCR and real time PCR methods using primer designed from rat alpha-1-microglobulin gene were performed to amplify cDNA in cellular samples. Western blot was used to monitor the amount of specific proteins produced in culture media and in cells. The result showed that cellular AMBP gene was up-regulated and alpha-1-microglobulin protein increased in culture media after treated with oxalate and calcium oxalate crystal.