陳慧玲2006-07-262018-07-112006-07-262018-07-112001http://ntur.lib.ntu.edu.tw//handle/246246/22857背景： BSEP及FIC1基因異常近年來被發現是進 行性家族性肝內膽汁滯留的致病原因，這些基因的 變異在亞洲兒童並未被報告過，在前一年的研究中 我們首先發現了台灣兒童之散發性進行性肝內膽 汁滯留，約有80%是由BSEP及FIC1造成。本計畫針 對新生兒肝炎病患，探討BSEP及FIC1gene的角色， 同時了解在不同年齡，膽小管傳送蛋白的表現差異 情形。以釐清BSEP及FIC1在嬰兒進一步的致病角 色。 進行方法 :本計畫第第一部分部分探討新生兒特 別容易產生黃疸的原因，可能是因發育上與成人有 異而造成，因此利用胎兒，嬰兒，與成人的肝臟組 織切片，以半定量RT-PCR法，比較在不同年齡發育 上的差異，同時分析新生兒血液中膽酸濃度。第二 部分對新生兒肝炎病人之cDNA做BSEP及FIC1 基因 序列分析，以找出基因變異在新生兒肝炎之角色。 結果：本實驗發現BSEP 及FIC1 等肝細胞傳送蛋白 的表現，在人類胚胎發育的第16 周已有明顯表現， 在20 周以後到達明顯的程度，與胎兒開始分泌膽 汁的時期配合。而在出生後的新生嬰兒，約在四個 月到達最高程度，成人的表現量比胎兒低。第二部 分測定新生兒肝炎全段序列，共完成五例BSEP 及 三例FIC1 序列鑑定，均未發現mutation。新生兒 肝炎病例之BSEP 免疫染色均為陽性，顯示在嬰兒 兒肝內膽汁滯留中BSEP 的表現為陽性或加強。 討論：本研究提供了在人類胎兒及新生兒至成人的 肝內傳送蛋白的發育情形，證實BSEP 等基因在胎 兒已有表現，並且在出生後迅速增加。而在一般的 新生兒肝炎病患，亦有BSEP 的表現，而大多沒有 BSEP 的基因變異。Background/Aim: FIC1 and BSEP mutations have recently been found to cause progressive familial intrahepatic cholestasis (PFIC). Mutations in both genes have not been reported in Asian children. We had identified novel mutations in FIC1 and BSEP genes in 80% of Taiwanese infants with sporadic progressive intrahepatic cholestasis. This study is to elucidate the different expression levels of BSEP and other canalicular transporters in different fetal and postnatal ages, and to determine the role of FIC1 and BSEP mutations in children with neonatal hepatitis in Taiwan. Methods and Results: In the first part we examined the expression levels of fetal, neonatal, and adult livers by semi-quantitative RT-PCR. We found that BSEP and other transporters were expressed as early as 16 weeks of gestational age, and became strongly expressed after 20 weeks. This finding is compatible with the time when bile starts to flow from liver to intestine. We also determined the serum bile acid levels in normal newborn infants and found that normal newborn have serum bile acid below 94mmole/L, much higher that adults, which was below 10 mmole/L. In the second part we sequenced the entire coding sequences of BSEP in 5 patients and FIC1 in 2 patients with neonatal hepatitis. We found no mutation in these patients. Immunohistochemical staining in the liver tissue of neonatal hepatitis patients showed positive results. Discussion: Our study provided the first data on the expression levels of liver canalicular transporters in human fetal and postnatal periods. This data helps to elucidate the neonatal bile physiology. In addition, we found no evidence that genetic alternations occurred in benign neonatal hepatitis.application/pdf74218 bytesapplication/pdfzh-TW國立臺灣大學醫學院小兒科膽汁滯留基因變異兒童肝病胚胎發育cholestasisgene mutationchildhood liver diseasefetal development[SDGs]SDG3reporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/22857/1/892314B002401.pdf