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大鼠肝脏中低Km和高Km醛脱氢酶活性腺泡内分布图谱的微量定量测定

Microquantitative determination of intra-acinar distribution profiles of low-Km and high-Km aldehyde dehydrogenase activity in rat liver.

作者信息

Maly I P, Sasse D

机构信息

Anatomisches Institut der Universität Basel, Switzerland.

出版信息

Eur J Biochem. 1987 Dec 30;170(1-2):173-8. doi: 10.1111/j.1432-1033.1987.tb13683.x.

Abstract

Microquantitative measurements of total and of low-Km aldehyde dehydrogenase (ALDH) activity with millimolar and micromolar concentrations of acetaldehyde and propionaldehyde were carried out on the livers of male and female rats. Lyophilized cryostat sections of liver parenchyma were microdissected along the entire sinusoidal length from the terminal afferent vessels to the terminal efferent venule. ALDH activity was measured in a microbiochemical assay using the oil-well technique with luminometric determination of NADH. On the basis of single measurements, mean values of total, low-Km and high-Km ALDH activity could be calculated and the specific distribution patterns graphically demonstrated. The two substrates acetaldehyde and propionaldehyde yielded similar values of ALDH activity, the intraacinar distribution profiles of which showed characteristic sex differences. In the liver of the male rat high-Km ALDH activity has two flat peaks in the periportal and the perivenous area, while low-Km ALDH activity is almost evenly distributed throughout the acinus. In the livers of female rats, both high-Km and low-Km ALDH activity shows a continuous gradient which decreases from the periportal to the perivenous zone (pp/pv = 1.4:1). It was therefore possible to demonstrate that the maxima of alcohol dehydrogenase activity and of low-Km ALDH activity are localized in opposite parts of the liver acinus of the female rat. This heterotopy should have consequences with respect to hepatotoxicity after alcohol ingestion.

摘要

使用毫摩尔和微摩尔浓度的乙醛和丙醛,对雄性和雌性大鼠的肝脏进行了总醛脱氢酶(ALDH)和低Km醛脱氢酶活性的微量定量测量。将肝脏实质的冻干低温切片沿着从终末传入血管到终末传出小静脉的整个窦状长度进行显微解剖。采用油井技术和NADH的发光测定法,通过微生物化学分析测量ALDH活性。基于单次测量,可以计算出总ALDH、低Km和高Km ALDH活性的平均值,并以图形方式展示特定的分布模式。两种底物乙醛和丙醛产生了相似的ALDH活性值,其腺泡内分布曲线显示出特征性的性别差异。在雄性大鼠的肝脏中,高Km ALDH活性在门静脉周围和静脉周围区域有两个平坦的峰值,而低Km ALDH活性几乎均匀地分布在整个腺泡中。在雌性大鼠的肝脏中,高Km和低Km ALDH活性均呈现出从门静脉周围到静脉周围区域逐渐降低的连续梯度(门静脉周围/静脉周围 = 1.4:1)。因此,可以证明乙醇脱氢酶活性和低Km ALDH活性的最大值位于雌性大鼠肝腺泡的相反部位。这种异位现象在摄入酒精后对肝毒性方面应该会产生影响。

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