Long-term maintenance of the adult pattern of liver-specific expression for P-450b, P-450e, albumin and alpha-fetoprotein genes in intrasplenically transplanted hepatocytes.
Long-term maintenance of the adult pattern of liver-specific expression for P-450b, P-450e, albumin and alpha-fetoprotein genes in intrasplenically transplanted hepatocytes.
复制标题
脾内移植肝细胞中 P-450b、P-450e、白蛋白和甲胎蛋白基因的肝脏特异性表达的成人模式的长期维持。
DOI：
10.1002/hep.1840110410
复制
发表时间：
1990
期刊：
影响因子：
--
通讯作者：
Gumucio,JJ
中科院分区：
文献类型：
--
作者：
Maganto,P;Traber,PG;Rusnell,C;Dobbins3rd,WO;Keren,D;Gumucio,JJ
从Fischer 344大鼠的肝脏中分离出肝细胞，并将其移植到同一品系的大鼠的脾中，在没有免疫抑制药物的情况下至少存活了15个月。肝细胞只附着在脾的红髓上，最常见的是成团的，没有明显的结构。在15个月的时间里，脾内移植的肝细胞表达细胞色素P-450B、P-450E和白蛋白信使RNA，而甲胎蛋白信使RNA不表达。此外，白蛋白和P-450基因的相对表达与肝脏相似。例如，白蛋白信使RNA的表达水平高于P-450b或e信使RNA。与P-450B或P-450E特异性寡核苷酸杂交的Northern杂交结果表明，在肝脏中，P-450B和P-450E基因均被苯巴比妥诱导。在肝细胞移植后15天、1、6和15个月进行定量狭缝印迹杂交表明，单剂量苯巴比妥可诱导细胞色素P-450B和P-450E信使RNA增加约20~30倍。这种诱导水平也与在肝脏中观察到的相似。因此，脾内移植的肝细胞是一个独特的系统，在这种系统中，在肝外活体环境中培养的肝细胞，至少在15个月内保持这四个肝基因类似于成人肝脏的表达模式。此外，这些研究表明，成年肝细胞对P-450B和P-450E基因诱导的苯巴比妥的反应，既不需要肝脏组织成腺泡，也不需要特定的带状血窦微环境。
Hepatocytes isolated from livers of Fischer 344 rats and transplanted into the spleens of rats from the same strain survived for at least 15 mo in the absence of immunosuppressive drugs. Hepatocytes attached themselves only in the red pulp of the spleen, most commonly in clumps without a discernible structure. Throughout the 15‐mo period, intrasplenically transplanted hepatocytes expressed cytochrome P‐450b, P‐450e and albumin messenger RNAs, whereas alphafetoprotein messenger RNA was not expressed. In addition, the relative expression of albumin and P‐450 genes was similar to that in liver. For example, albumin messenger RNA was expressed to higher levels than P‐450b or e messenger RNAs. Northern blots hybridized with oligonucleotides specific for P‐450b or P‐450e showed that, as in liver, both P‐450b and P‐450e genes were induced in response to phenobarbital. Quantitative slot‐blot hybridizations performed at 15 days and 1, 6, and 15 mo after hepatocyte transplantation revealed that cytochrome P‐450b and P‐450e messenger RNAs were induced about 20‐ to 30‐fold by a single dose of phenobarbital. This level of induction was also similar to that observed in liver. Hence, intrasplenically transplanted hepatocytes represent a unique system in which hepatocytes, cultured in an extrahepaticin vivoenvironment, maintain for at least 15 mo a pattern of expression for these four liver genes similar to that in the adult liver. Moreover, these studies suggest that neither the organization of liver into acini nor a specific zonal sinusoidal microenvironment is necessary for adult hepatocytes to respond to phenobarbital with induction of P‐450b and P‐450e genes.(HEPATOLOGY 1990; 11:585:593.)