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大鼠胰岛中的阿片肽。免疫反应性甲硫氨酸脑啡肽、亮氨酸脑啡肽和BAM - 22P。

Opioid peptides in rat islets of Langerhans. Immunoreactive met- and leu-enkephalins and BAM-22P.

作者信息

Timmers K I, Voyles N R, King C, Wells M, Fairtile R, Recant L

出版信息

Diabetes. 1986 Jan;35(1):52-7. doi: 10.2337/diab.35.1.52.

Abstract

Previous studies have shown that met- and leu-enkephalins are present in extracts of whole pancreas obtained from guinea pigs and human cadavers. The present studies demonstrate that immunoreactive methionine (met)- and leucine (leu)-enkephalins present in rat pancreas are localized in islets of Langerhans. Immunohistochemical staining of fixed, whole pancreas indicated that only islet endocrine cells were heavily stained when any of four different met- and leu-enkephalin-directed antisera or an anti-BAM-22P (bovine adrenal medulla docosapeptide) antiserum was used. The peptides were characterized by a combination of gel-filtration chromatography, high-performance liquid chromatography (HPLC), and specific radioimmunoassay. Free met-enkephalin content in extracts of rat islets was 90-fold enriched over content in extracts of whole pancreas (1.72 +/- 0.35 versus 0.019 +/- 0.007 pmol/mg protein). Treatment with trypsin and carboxy-peptidase-B of high-molecular-weight peptides extracted from pancreas or islets resulted in release of additional met-enkephalin immunoreactivity, which was 39-fold enriched in islets compared with pancreas (5.90 +/- 0.58 and 0.153 +/- 0.032 pmol/mg protein, respectively). Total islet content (per milligram protein) of met-enkephalin-containing peptides was similar to that reported elsewhere for bovine hypothalamus. The immunohistochemical data as well as the enrichment of extractable enkephalins in islets compared with whole pancreas indicate that essentially all the met-enkephalin present in pancreas is localized in islets, while the presence of BAM-22P immunoreactivity in islets is consistent with biosynthesis of enkephalins in islet cells via a preprohormone, such as that described in the bovine adrenal medulla and rat brain.

摘要

先前的研究表明,甲硫氨酸脑啡肽和亮氨酸脑啡肽存在于从豚鼠和人类尸体获取的全胰腺提取物中。目前的研究证明,大鼠胰腺中存在的免疫反应性甲硫氨酸(met)-和亮氨酸(leu)-脑啡肽定位于胰岛。对固定的全胰腺进行免疫组织化学染色表明,当使用四种不同的针对met-和leu-脑啡肽的抗血清或抗BAM-22P(牛肾上腺髓质二十二肽)抗血清中的任何一种时,只有胰岛内分泌细胞被重度染色。这些肽通过凝胶过滤色谱、高效液相色谱(HPLC)和特异性放射免疫测定相结合的方法进行鉴定。大鼠胰岛提取物中的游离甲硫氨酸脑啡肽含量比全胰腺提取物中的含量富集了90倍(分别为1.72±0.35和0.019±0.007 pmol/mg蛋白质)。用胰蛋白酶和羧肽酶B处理从胰腺或胰岛中提取的高分子量肽,会导致额外的甲硫氨酸脑啡肽免疫反应性释放,与胰腺相比,胰岛中的这种反应性富集了39倍(分别为5.90±0.58和0.153±0.032 pmol/mg蛋白质)。含甲硫氨酸脑啡肽的肽的胰岛总含量(每毫克蛋白质)与其他地方报道的牛下丘脑的含量相似。免疫组织化学数据以及与全胰腺相比胰岛中可提取脑啡肽的富集情况表明,胰腺中存在的基本上所有甲硫氨酸脑啡肽都定位于胰岛,而胰岛中BAM-22P免疫反应性的存在与胰岛细胞中通过前激素原生物合成脑啡肽一致,如在牛肾上腺髓质和大鼠脑中所描述的那样。

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