Défize J, Pals G, Frants R R, Westerveld B D, Meuwissen S G, Eriksson A W
Prog Clin Biol Res. 1985;173:147-57.
De novo synthesis of pepsinogens was demonstrated in gastric glands isolated from rabbit and human gastric mucosa. The isolated glands were incubated in an amino acid free, Minimum Eagles Medium supplemented with a 14C labelled amino acid mixture. The glands were centrifuged at different time intervals and aliquots of gland homogenates and medium were run on polyacrylamide slab gels. Newly synthesized pepsinogens were demonstrated by autoradiography. Incorporation of 14C was detected after 30 minutes of culture and increased almost linearily with time over 4 h. By comparing the electrophoretic patterns after autoradiography, protein and pepsinogen activity staining, it was concluded that the glands synthesize mainly pepsinogens. Cimetidine, at a concentration of 10(-4)M strongly inhibited pepsinogen synthesis. Spontaneous secretion of pepsinogens into the medium was very low and relatively constant. Cyclic dibutyryl AMP markedly stimulated the secretion of pepsinogens into the medium. The results show, that isolated gastric glands are capable of synthesis and secretion of pepsinogens and that each function can be stimulated and inhibited selectively.
在从兔和人胃黏膜分离出的胃腺中证实了胃蛋白酶原的从头合成。将分离出的胃腺在不含氨基酸的最低限度伊格尔培养基中培养,并添加14C标记的氨基酸混合物。在不同时间间隔对胃腺进行离心,取胃腺匀浆和培养基的等分试样在聚丙烯酰胺平板凝胶上进行电泳。通过放射自显影法证实了新合成的胃蛋白酶原。培养30分钟后检测到14C的掺入,并且在4小时内几乎呈线性增加。通过比较放射自显影后的电泳图谱、蛋白质和胃蛋白酶原活性染色,得出胃腺主要合成胃蛋白酶原的结论。浓度为10(-4)M的西咪替丁强烈抑制胃蛋白酶原的合成。胃蛋白酶原向培养基中的自发分泌非常低且相对恒定。环二丁酰腺苷酸显著刺激胃蛋白酶原向培养基中的分泌。结果表明,分离出的胃腺能够合成和分泌胃蛋白酶原,并且每种功能都可以被选择性地刺激和抑制。