二酰基甘油和蛋白激酶C通路不参与原代大鼠肝细胞的胰岛素信号传导。

Irmelin Probst, Ulrich Beuers, Birgit Drabent, Kirsten Unthan-Fechner, Peter Bütikofer
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引用次数: 6

摘要

二酰基甘油(DAG)和蛋白激酶C (PKC)亚型与肌肉和脂肪细胞中的胰岛素信号传导有关。我们评估了用地塞米松培养的成年大鼠肝细胞中DAG和PKC参与胰岛素的作用,但在没有血清的情况下,48小时。我们的结果表明,尽管胰岛素刺激糖酵解和糖原合成,但它对DAG质量或分子种类组成没有影响。表皮生长因子对糖酵解表现出预期的胰岛素模拟作用,而ATP和外源磷脂酶C则作为拮抗剂并消除胰岛素信号。与胰岛素相似,表皮生长因子对DAG质量和分子种类组成没有影响。相比之下,ATP和磷脂酶C均诱导了几个DAG分子种类的显著增加,包括18:0/20:4,18:0/20:5,18:0/22:5,18:1/18:1的减少。这些变化与磷脂酶D活性的增加是平行的,这在胰岛素处理的细胞中是不存在的。通过免疫印迹或PKC活性测定,我们发现胰岛素和ATP都不能将PKCalpha、-delta、-epsilon或-zeta同工型从细胞质转移到膜上,在培养6或48小时的细胞中。同样,胰岛素对免疫沉淀PKCzeta没有影响。双吲哚酰马来酰亚胺可完全减轻12-肉豆蔻酸酯对糖原性胰岛素信号的抑制,而不是ATP对糖原性胰岛素信号的抑制。最后,胰岛素对灌注肝脏中DAG质量或PKC异构体的易位没有影响,尽管它使胰高血糖素刺激的葡萄糖输出减少了75%。这些结果表明,磷脂酶C和D或多种PKC亚型不参与肝脏胰岛素信号链。
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The diacylglycerol and protein kinase C pathways are not involved in insulin signalling in primary rat hepatocytes.

Diacylglycerol (DAG) and protein kinase C (PKC) isoforms have been implicated in insulin signalling in muscle and fat cells. We evaluated the involvement of DAG and PKC in the action of insulin in adult rat hepatocytes cultured with dexamethasone, but in the absence of serum, for 48 h. Our results show that although insulin stimulated glycolysis and glycogen synthesis, it had no effect on DAG mass or molecular species composition. Epidermal growth factor showed the expected insulin-mimetic effect on glycolysis, whereas ATP and exogenous phospholipase C acted as antagonists and abolished the insulin signal. Similarly to insulin, epidermal growth factor had no effect on DAG mass or molecular species composition. In contrast, both ATP and phospholipase C induced a prominent increase in several DAG molecular species, including 18:0/20:4, 18:0/20:5, 18:0/22:5 and a decrease in 18:1/18:1. These changes were paralleled by an increase in phospholipase D activity, which was absent in insulin-treated cells. By immunoblotting or by measuring PKC activity, we found that neither insulin nor ATP translocated the PKCalpha, -delta, -epsilon or -zeta isoforms from the cytosol to the membrane in cells cultured for six or 48 h. Similarly, insulin had no effect on immunoprecipitable PKCzeta. Suppression of the glycogenic insulin signal by phorbol 12-myristate 13-acetate, but not by ATP, could be completely alleviated by bisindolylmaleimide. Finally, insulin showed no effect on DAG mass or translocation of PKC isoforms in the perfused liver, although it reduced the glucagon-stimulated glucose output by 75%. Together these results indicate that phospholipases C and D or multiple PKC isoforms are not involved in the hepatic insulin signal chain.

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