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大鼠表皮生长因子:纯化及组织含量

Rat epidermal growth factors: purification and tissue content.

作者信息

Sakamoto K, Oka Y, Uda K, Chiba K, Waki Y, Suzuki W, Itoh K, Aburada M, Fujiwara K, Wakabayashi K

机构信息

Tsumura Research Institute for Pharmacology, Inashiki, Japan.

出版信息

Endocrinol Jpn. 1989 Apr;36(2):289-97. doi: 10.1507/endocrj1954.36.289.

Abstract

Rat epidermal growth factor (rEGF) was isolated from the submaxillary gland of male rat by reversed phase high performance liquid chromatography and ion exchange chromatography. The binding of purified rEGF to human carcinoma cells (A-431) and its tritiated thymidine uptake on rat epidermal fibroblast cells (FR) were almost the same as those of purified commercially available mouse EGF (mEGF). Antisera to rEGF was raised in rabbits and a radioimmunoassay (RIA) system was established. The assay range of the RIA was about 1.0 to 100 ng/ml. The within assay coefficients of variation were 5 to 10%, while the between assay coefficients of variation were 5 to 13%. The tissue content of rEGF of male rats (10 weeks old) was examined. As a result, the submaxillary gland was found to contain a very high concentration of rEGF (214 micrograms/g wet tissue) as predicted, and digestive tissues, stomach, intestine and duodenum contained 2.49, 3.57, 9.44 ng/g wet tissue, respectively. The amounts in prostate and seminal vesicle were relatively high, being 65.6 and 2,268 ng/g wet tissue, respectively. The amount in the submaxillary gland increased markedly after 7 weeks of age. These results suggest that EGF is an important factor in gonadal function.

摘要

通过反相高效液相色谱法和离子交换色谱法从雄性大鼠的颌下腺中分离出大鼠表皮生长因子(rEGF)。纯化的rEGF与人癌细胞(A-431)的结合及其对大鼠表皮成纤维细胞(FR)的氚化胸腺嘧啶摄取情况与市售纯化小鼠表皮生长因子(mEGF)几乎相同。用兔制备了抗rEGF血清,并建立了放射免疫分析(RIA)系统。该RIA的检测范围约为1.0至100 ng/ml。批内变异系数为5%至10%,批间变异系数为5%至13%。检测了10周龄雄性大鼠的rEGF组织含量。结果发现,颌下腺如预期含有非常高浓度的rEGF(214微克/克湿组织),消化组织、胃、小肠和十二指肠分别含有2.49、3.57、9.44纳克/克湿组织。前列腺和精囊中的含量相对较高,分别为65.6和2268纳克/克湿组织。颌下腺中的含量在7周龄后显著增加。这些结果表明,表皮生长因子是性腺功能中的一个重要因素。

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