臺灣博碩士論文加值系統

甲一型微球蛋白(alpha-1-microglobulin)是一種與bikunin蛋白由同段基因(AMBP gene: □-1m-bikunin precursor gene)一起產生後，於細胞中分解而形成的蛋白質，大多在肝臟中製造，而後經由腎臟排出到尿液中。甲一型微球蛋白與bikunin蛋白皆被認為有抑制尿路結石生長的功能。本實驗利用草酸分子及草酸鈣經過的不等時間剌激近曲腎小管細胞(LLC-PK1)，再以分子生物學技術萃取細胞中的核醣核酸，反轉錄成互補段的去氧核醣核酸，並利用甲一型球蛋白的引體進行聚合酶反應及即時聚合酶反應，以擴大產物總量，與原始互補段的去氧核醣核酸的總量比較，反推出原有細胞中AMBP信息核醣核酸的多寡。利用西方墨點法檢視蛋白質的生成，結果發現經過草酸及草酸鈣剌激後，信息核醣核酸及蛋白質均有增加的現象。

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.

目錄
壹、中文摘要-------------------------------------------------------2
貳、英文摘要 -----------------------------------------------------3
參、緒論 -----------------------------------------------------------5
肆、材料及方法 --------------------------------------------------8
伍、結果 -----------------------------------------------------------13
陸、討論 -----------------------------------------------------------16
柒、結論 -----------------------------------------------------------20
捌、參考文獻-----------------------------------------------------21
玖、圖 --------------------------------------------------------------25

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