INCREASED SODIUM-ION INFLUX IS NECESSARY TO INITIATE RAT HEPATOCYTE PROLIFERATION
INCREASED SODIUM-ION INFLUX IS NECESSARY TO INITIATE RAT HEPATOCYTE PROLIFERATION
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DOI：
10.1016/0092-8674(79)90364-7
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发表时间：
1979-01-01
期刊：
影响因子：
64.5
通讯作者：
LEFFERT, HL
中科院分区：
文献类型：
--
作者：
KOCH, KS;LEFFERT, HL
含有10-50 ng胰岛素、胰高血糖素和表皮生长因子（EGF）ml-1的无血清培养基刺激10-15日龄原代肝细胞培养物中的成年大鼠肝细胞增殖。这种反应的动力学模拟了67%肝切除术后伴随肝再生的肝细胞转变。阿米洛利，一种Na+内流抑制剂，在体外可逆地阻断这些转变（ID 50 apprx. 0.02 mM）和体内（ID 50. 25 mg kg-1）。用其它阳离子通量调节剂，包括哇巴因（ID 50 apprx. 0.2 mM）、0.2 μ M莫能菌素和0.2 μ M尼日利亚菌素，但不与0.3mM呋塞米或河豚毒素一起使用。培养物中的预复制间隔（0-12 h）的特征在于对EGF（0-3 h）的优先细胞反应性，随后是胰岛素加胰高血糖素（3-12 h）。平行培养和动物研究表明，阿米洛利敏感和复制前的时间间隔相吻合。在文化中，突发的22 Na+内流，刺激肽补充培养基在1分钟内，但在12小时后减少，是由阿米洛利延迟。该药物还阻断延迟的复制前事件，包括在4-8小时增加氨基酸A转运系统功能，以及在8-12小时分别将3 H-尿苷和3 H-亮氨酸掺入RNA和蛋白质。至少有2个时间顺序的过程是必要的，以启动肝脏生长完全：激活Na+通量系统的肽类似或相同的EGF;和增强这些和随后的细胞事件的联合行动的胰岛素加胰高血糖素。
Serum-free media containing 10-50 ng insulin, glucagon and epidermal growth factor (EGF) ml-1 stimulate adult rat hepatocyte proliferation in 10-15 day old primary liver cell cultures. The kinetics of this response simulate hepatocellular transitions that accompany liver regeneration after 67% hepatectomy. Amiloride, a Na+ influx inhibitor, reversibly blocks these transitions in vitro (ID50 .apprx. 0.02 mM) and in vivo (ID50 .apprx. 25 mg kg-1). Inhibition is observed with other cation flux modulators, including ouabain (ID50 .apprx. 0.2 mM), 0.2 .mu.M monensin and 0.2 .mu.M nigericin, but not with 0.3 mM furosemide or tetrodotoxin. The prereplicative interval in culture (0-12 h) is characterized by preferential cellular responsiveness to EGF (0-3 h) followed by insulin plus glucagon (3-12 h). Parallel culture and animal studies show that the amiloride-sensitive and prereplicative intervals coincide. In culture, a burst of 22Na+ influx, stimulated by peptide-supplemented media within 1 min but decreased later at 12 h, is retarded by amiloride. This drug also blocks delayed prereplicative events involving increased amino acid A transport system function at 4-8 h, and 3H-uridine and3H-leucine incorporation into RNA and protein, respectively, at 8-12 h. At least 2 time-ordered processes are necessary to initiate hepatic growth fully: activation of Na+ flux systems by peptides similar or identical to EGF; and potentiation of these and subsequent cellular events by the combined action of insulin plus glucagon.