Tabuchi Y, Sugiyama N, Horiuchi T, Furuhama K, Furusawa M
Molecular Biology Research Laboratory, Daiichi Pharmaceutical Co., Ltd., Tokyo, Japan.
Digestion. 1997;58(1):28-33. doi: 10.1159/000201420.
The mechanism of regulation of mucus production in the gastric mucosa remains unclear. Recently, we established a gastric surface mucous cell line GSM06, which produces periodic acid-Shiff (PAS)-positive glycoconjugate (mucus) layers on the cell surface, from transgenic mice harboring a temperature-sensitive simian virus 40 large T-antigen gene. In this study, GSM06 cells were examined for its production of PAS-positive glycoconjugate layers to acid secretagogues and growth factors. The cells were cultured at nonpermissive temperature (39 degrees C) for 3-18 days and stained with PAS. Insulin (1-30 microg/ml; 0.29-8.6 microM) time- and dose-dependently increased production of glycoconjugates on the cell surface. When glycoconjugate layers produced by stimulation of insulin (3-30 microg/ml; 0.86-8.6 microM) were removed from the cell surface of GSM06 cells by a mild trypsin treatment, PAS-positive materials were remarkably decreased (day 18). In addition, morphological findings indicate that a high concentration of insulin (30 microg/ml; 8.6 microM) produced thick PAS-positive glycoconjugate layers just like normal gastric surface mucosa on the cell surface on day 18. In contrast, histamine (0.1-100 microM), carbachol (0.1-100 microM), gastrin-17 (0.1-100 nM), epidermal growth factor (0.01-10 ng/ ml; 1.7-1,700 pM), transforming growth factor-alpha (0.01-10 ng/ml; 1.8-1,800 pM), and fetal bovine serum (1-10%) did not increase glycoconjugate production. These findings suggest that insulin is a stimulator of glycoconjugate production, and stimulates production of glycoconjugate layers on the cell surface in the gastric surface mucous cell line GSM06.
胃黏膜中黏液产生的调节机制仍不清楚。最近，我们从携带温度敏感型猿猴病毒40大T抗原基因的转基因小鼠中建立了一种胃表面黏液细胞系GSM06，该细胞系在细胞表面产生过碘酸希夫（PAS）阳性糖缀合物（黏液）层。在本研究中，检测了GSM06细胞对酸促分泌剂和生长因子产生PAS阳性糖缀合物层的情况。将细胞在非允许温度（39℃）下培养3 - 18天，并用PAS染色。胰岛素（1 - 30微克/毫升；0.29 - 8.6微摩尔）能时间和剂量依赖性地增加细胞表面糖缀合物的产生。当通过温和的胰蛋白酶处理从GSM06细胞表面去除由胰岛素（3 - 30微克/毫升；0.86 - 8.6微摩尔）刺激产生的糖缀合物层时，PAS阳性物质显著减少（第18天）。此外，形态学结果表明，高浓度胰岛素（30微克/毫升；8.6微摩尔）在第18天在细胞表面产生了像正常胃表面黏膜一样厚的PAS阳性糖缀合物层。相比之下，组胺（0.1 - 100微摩尔）、卡巴胆碱（0.1 - 100微摩尔）、胃泌素 - 17（0.1 - 100纳摩尔）、表皮生长因子（0.01 - 10纳克/毫升；1.7 - 1700皮摩尔）、转化生长因子 - α（0.01 - 10纳克/毫升；1.8 - 1800皮摩尔）和胎牛血清（1 - 10%）均未增加糖缀合物的产生。这些发现表明胰岛素是糖缀合物产生的刺激物，并能刺激胃表面黏液细胞系GSM06细胞表面糖缀合物层的产生。
