Gastrin effects on isolated rat enterochromaffin-like cells in primary culture.
Gastrin effects on isolated rat enterochromaffin-like cells in primary culture.
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胃泌素对原代培养中分离的大鼠肠嗜铬样细胞的影响。
DOI：
10.1152/ajpgi.1994.267.4.g663
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
Sachs,G
中科院分区：
文献类型：
--
作者：
Prinz,C;Scott,DR;Hurwitz,D;Helander,HF;Sachs,G
激素胃泌素通过从胃肠嗜铬样（ECL）细胞释放组胺刺激胃酸分泌，并诱导体内ECL细胞增殖。本研究使用> 90%纯ECL细胞制备物在培养中比较胃泌素对组胺释放、组氨酸脱羧酶（HDC）活性和DNA合成的作用。在孵育5 min内，胃泌素和八肽胆囊收缩素（CCK-8，非硫酸化）诱导ECL细胞（原代培养24-96 h）释放组胺[激发50%最大反应（EC 50）的浓度分别为4和2 x 10（-11）M]。CCK-B拮抗剂L-365，260可抑制这种作用[抑制50%最大反应的浓度（IC 50），2 × 10（-8）M]，而CCK-A拮抗剂L-364，718（10（-8）M）和酪氨酸激酶抑制剂genistein（10（-4）M）则无作用。组胺释放与细胞内Ca 2+的双相升高相关。孵育60分钟后，胃泌素刺激HDC活性2 - 3倍（EC 50，10（-10）M）。胃泌素还增加ECL细胞中的DNA合成，通过掺入5-溴-2 '-脱氧尿苷（BrdU）测量的EC 50为1.7 x 10（-12）M。孵育48-96 h后，高达20%的ECL细胞的阳性核免疫染色增加了2 - 3倍。L-365，260（IC 50，5 × 10（-9）M）和染料木素（10（-4）M）可抑制该作用，但L-364，718（10（-8）M）未改变该作用。抗分泌药物奥美拉唑，兰索拉唑，泮托拉唑不影响BrdU掺入分离的ECL细胞。总之，急性和慢性胃泌素对ECL细胞的作用是通过CCK-B受体介导的，但在表观受体亲和力和信号转导途径上有所不同。
The hormone gastrin stimulates acid secretion by releasing histamine from gastric enterochromaffin-like (ECL) cells and induces ECL cell proliferation in vivo. This study uses a > 90% pure ECL cell preparation in culture to compare gastrin effects on histamine release, histidine decarboxylase (HDC) activity, and DNA synthesis. Gastrin and the cholecystokinin octapeptide (CCK-8, nonsulfated) induced histamine release from ECL cells (24-96 h of primary culture) within 5 min of incubation [concentration eliciting 50% of maximal response (EC50), 4 and 2 x 10(-11) M, respectively]. The CCK-B antagonist L-365,260 inhibited this effect [concentration inhibiting 50% of maximal response (IC50), 2 x 10(-8) M], whereas the CCK-A antagonist L-364,718 (10(-8) M) and the tyrosine kinase inhibitor genistein (10(-4) M) had no effect. Histamine release was associated with a biphasic elevation of intracellular Ca2+. Gastrin stimulated HDC activity two- to threefold after 60 min of incubation (EC50, 10(-10) M). Gastrin also increased DNA synthesis in ECL cells, with an EC50 of 1.7 x 10(-12) M as measured by the incorporation of 5-bromo-2'-deoxyuridine (BrdU). Positive nuclear immunostaining increased two- to threefold in up to 20% of ECL cells after 48-96 h of incubation. This effect was inhibited by L-365,260 (IC50, 5 x 10(-9) M) and by genistein (10(-4) M) but was not altered by L-364,718 (10(-8) M). The antisecretory drugs omeprazole, lansoprazole, and pantoprazole did not affect BrdU incorporation in isolated ECL cells. In conclusion, acute and chronic gastrin effects on the ECL cell are mediated via CCK-B receptors but differ in apparent receptor affinity and signal transduction pathways.