ANALYSIS OF THE BILE ACID PROFILES AND APPLICATION TO THE CONGENITAL LIVER DISORDER

胆汁酸谱分析及其在先天性肝病中的应用

• 批准号: 02557089
• 负责人: TOHMA Masahiko
• 金额: $ 3.07万
• 依托单位: HIGASHI-NIPPON-GAKUEN UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02557089/
• 关键词: FETAL BILE ACID; BILE ACID BIOSYNTHETIC INTERMEDIATE; NEONATAL LIVER DESEASE; BILE ACID PROFILE; CAPILLARY GAS CHROMATOGRAPHY; CONGENITAL BILIARY ATRESIA; IMMUNOASSAY; ZELLWEGER SYNDROME; 先天性肝胆道疾患; 抱合型胆汁酸; 酵素免疫測定法; マススクリーニング法; C_<27>-胆汁酸; マスス

The establishment of the diagnosis of congenital liver disorder is urgently required for the remedy of liver disorder in early life. The aim of this research is the development of quantitative analysis of fetal bile acids by GC-MS and immunological assay, elucidation of the change of bile acid profiles with growth, and establishment of the precise diagnosis and masscreening of congenital biliary atresia.(1) The stereoisomeric C_<27>-bile acids and bile alcohols were chemically synthesized for the authentic specimens. The newly developed analysis of these intermediates was achieved by capillary gas chromatography.(2) Highly sensitive radioimmunoassay and enzyme immunoassay were developed. By the established method, the change of conjugated fetal bile acids in urine of pregnant women and newborns was examined.(3) The bile acid composition of neonatal periods changed with growth. In the case of preterm infants, the similar results were observed. The result of comparison of the ratio of 1beta-hydroxycholic acid to cholic acid indicated the activity of 1beta-hydroxylase decreased with growth.(4) 1. Diagnostic analysis of congenital biliary atresia was accomplished by enzyme immunoassay. The possibility of masscreening using dried blood spots was also examined. 2. From the urine of Zellweger syndrome, THCA and varanic acid were identified and determined by GLC. 3. In addition to 1beta-hydroxycholic acid, 1beta-hydroxylated C_<23>- and C_<22>-bile acids were found in the urine of patients with cerebrotendinous xantomatosis.

建立先天性肝病的诊断标准是治疗生命早期肝病的迫切需要。本研究的目的是建立胎儿胆汁酸的GC-MS定量分析方法和免疫学分析方法，阐明胎儿胆汁酸谱随生长发育的变化规律，建立先天性胆道闭锁的准确诊断和筛查方法。(1)化学合成胆汁酸和胆汁醇的立体异构体。建立了高灵敏的放射免疫分析方法和酶免疫分析方法。用建立的方法检测孕妇和新生儿尿液中结合胆汁酸的变化。(3)新生儿期胆汁酸组成随生长发育而变化。在早产儿中，也观察到了类似的结果。比较1β-羟基胆酸/胆酸比值的结果表明，1β-羟基酶活性随生长发育而降低。(4)1.先天性胆道闭锁的诊断分析采用酶免疫法。还研究了使用干血斑点进行大规模筛查的可能性。2.从Zellweger综合征患者的尿液中，用气相色谱对THCA和呋喃酸进行了鉴定和含量测定。3.脑腱黄瘤患者尿液中除β-羟基胆酸外，还含有1β-羟基胆汁酸和C_(22)-胆汁酸。

项目成果

Kaoru Obinata, Hiroshi Nittono, Keijiro Yabuta, Reijiro Mahara, and Masahiko Tohma: "1beta-Hydroxylated Bile Acids in the Urine of Healthy Neonates." J. Pediatr. Gastroenterol. Nutr.15, (1). 1-5 (1992)

Kaoru Obinata、Hiroshi Nittono、Keijiro Yabuta、Reijiro Mahara 和 Masahiko Tohma：“健康新生儿尿液中的 1β-羟基胆汁酸”。

Hitoshi Chiba, Shigeo Ikegawa, Takao Kurosawa, Teruki Yoshimura, Yoshito Ito, Kazuhiko Matsuno, Kunihiko Kobayashi, and Masahiko Tohma: "A Direct Enzyme Immunoassay for 18-Hydroxycortisol in Urine: A Tool Good for Screening Primary Aldosteronism." J. Ster

Hitoshi Chiba、Shigeo Ikekawa、Takao Kurosawa、Teruki Yoshimura、Yoshito Ito、Kazuhiko Matsuno、Kunihiko Kobayashi 和 Masahiko Tohma：“尿液中 18-羟基皮质醇的直接酶免疫分析：筛查原发性醛固酮增多症的良好工具。”

Yoshinobu Hata: "Fetal Bile Acids in Congenital Biliary Atresis-Production in the Liver and Clinical Significance." Pediatr.Surg.Int.6. 309-312 (1991)

Yoshinobu Hata：“先天性胆道闭锁中的胎儿胆汁酸 - 肝脏中的产生及其临床意义。”

Shigeo Ikegawa: "Conjugated 1β-Hydroxycholic Acid in the Urine of Newborns and Pregnant Women measured by Radioimmunoassay using Antisera raised against N-(1β-Hydroxycholyl)-2-aminopropionic Accid-BSA Conjugate." Chem.Pharm.Bull.40. (1992)

Shigeo Ikekawa：“使用针对 N-(1β-羟基胆基)-2-氨基丙酸-BSA 结合物的抗血清，通过放射免疫测定法测量新生儿和孕妇尿液中的结合 1β-羟基胆酸，Chem.Pharm.Bull.40。” ）

Shigeo Ikegawa: "Determination of Conjugated Bile Acids in Human Urine by High-Performance Liquid Chromatography with Chemiluminescence Detection." J.Chromatogr.577. 229-238 (1992)

Shigeo Ikekawa：“采用化学发光检测的高效液相色谱法测定人尿液中的结合胆汁酸。”