Procollagen production by rat hepatocytes in primary culture.
Procollagen production by rat hepatocytes in primary culture.
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原代培养中大鼠肝细胞产生前胶原。
DOI：
10.1016/0304-4165(82)90037-x
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Seyer,JM
中科院分区：
文献类型：
--
作者：
Hatahara,T;Seyer,JM
肝细胞取自大鼠肝脏，并在原代培养物中维持长达 14 天。胶原蛋白合成在 3-5 天后达到最大，此后下降。胶原蛋白的产生率约为。同一动物的大鼠皮肤成纤维细胞传代 3-5 次后观察到的十分之一。 I 型原胶原蛋白是主要的大分子胶原蛋白种类，被鉴定为 450 000 道尔顿分子，该分子被转化为 120 000 道尔顿、变性、还原的原胶原链。预先用胃蛋白酶消化天然原胶原，释放出 95 000 道尔顿的胶原链，通过与载体大鼠皮肤 I 型胶原链共同迁移，鉴定为 α1(I) 和 α2(I)。 III 型原胶原的产生也通过 DEAE-纤维素色谱法进行了初步鉴定。这种材料是用针对牛皮肤 III 型前胶原的氨基末端延伸肽开发的类型特异性抗体进行分离和鉴定的。 I 型：III 型原胶原的相对分布估计为 7:3，与之前在整个大鼠肝脏中发现的比例相似。没有观察到 IV 型或 V 型原胶原生物合成的证据。这些结果表明，原代培养物中的大鼠肝细胞能够进行间质I型和III型胶原生物合成，其比例与其原位亲本肝组织中发现的比例相似。他们还表明，丰度较低的 IV 型（基底膜相关）或 V 型也不是这些细胞的主要胶原产物。
Hepatocytes were obtained from rat liver and maintained in primary culture for periods up to 14 days. Collagen synthesis was maximal after 3–5 days and declined thereafter. The rate of collagen production was appox. one-tenth that observed by the rat skin fibroblasts of the same animals after 3–5 passages. Type I procollagen, the major macromolecular collagenous species, was identified as a 450 000 dalton molecule which was converted to 120 000 dalton, denatured, reduced procollagen chains. Prior pepsin digestion of the native procollagen released 95 000 dalton collagen chains identified as α1(I) and α2(I) by co-migration with carrier rat skin type I collagen chains. The production of type III procollagen was also tentatively identified by DEAE-cellulose chromatography. This material was isolated and identified with type-specific antibodies developed against the amino-terminal extension peptide of bovine skin type III procollagen. The relative distribution of type I:type III procollagen was estimated at 7:3 similar to the ratio previously found in whole rat liver. No evidence of type IV or type V procollagen biosynthesis was observed. These results suggest that rat hepatocytes in primary culture are capable of interstitial type I and type III collagen biosynthesis in a ratio similar to that found in their parent hepatic tissue in situ. They also suggest that the less abundant type IV (basement membrane-associated) or type V are nor major collagenous products of these cells.