Protein Kinase C Inhibitors Enhance Differentiation of Rat Adipocyte Precursor Cells in Serum-Free Culture

Shinohara O., Murata Y., Kubota C., Shinagawa T.
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引用次数: 7

Abstract

The involvement of protein kinase C in differentiation of rat adipocyte precursor cells in serum-free culture was evaluated by using various protein kinase inhibitors. Induction of adipose conversion, which was maximal after 10 days of culture in the presence of 5 μg/ml insulin, 10 μg/ml transferrin, and 200 pM triiodothyronine, was inhibited by the addition of protein kinase C inhibitors, H-7 and staurosporine, in a dose dependent fashion with the maximal effect at 10 μM and 10 nM, respectively. Inhibition of adipocyte differentiation by 12-O-tetradecanoylphorbol 13-acetate (10−8M), an activator of protein kinase C, was reversed by a concomitant addition of either 10 μM H-7 or 10 nM staurosporine. HA1004, a potent inhibitor of cAMP- and cGMP-dependent protein kinases, with minimal inhibitory activity on protein kinase C, did not affect adipose conversion. Furthermore, H-89, another isoquinoline derivative with a selective inhibitory action on cAMP-dependent protein kinase, was without effect on cellular differentiation. These results indicate that the potentiation of adipogenesis by H-7 and staurosporine is mediated by suppression of protein kinase C and that protein kinase C is involved in adipocyte differentiation in an inhibitory fashion.

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蛋白激酶C抑制剂促进无血清培养大鼠脂肪前体细胞的分化
采用多种蛋白激酶抑制剂研究蛋白激酶C在无血清培养大鼠脂肪前体细胞分化中的作用。在胰岛素浓度为5 μg/ml、转铁蛋白浓度为10 μg/ml、三碘甲状腺原氨酸浓度为200 pM的条件下,脂肪转化在培养10 d后达到最大,而蛋白激酶C抑制剂H-7和staurosporine的加入抑制了脂肪转化,并呈剂量依赖性,分别在10 μM和10 nM处达到最大效果。蛋白激酶C的活化剂12-O-tetradecanoylphorbol 13-acetate(10−8M)对脂肪细胞分化的抑制作用被同时加入10 μM H-7或10 nM的staurosporine逆转。HA1004是一种cAMP-和cgmp依赖性蛋白激酶的有效抑制剂,对蛋白激酶C具有最小的抑制活性,不影响脂肪转化。此外,另一种对camp依赖性蛋白激酶具有选择性抑制作用的异喹啉衍生物H-89对细胞分化没有影响。这些结果表明,H-7和staurosporine对脂肪形成的增强是通过抑制蛋白激酶C介导的,蛋白激酶C以抑制方式参与脂肪细胞分化。
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