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目的探讨糖尿病相关因素──蛋白激酶C（PKC）对己糖激酶（hexokinases，HKs）的特异性调节以及与细胞对葡萄糖净利用的相关性，并探讨HKs在糖尿病肾病中的作用。方法SV40转基因鼠肾小球系膜细胞株（SV40MES13）作为细胞模型，分别使用6-磷酸葡萄糖脱氢酶耦联比色法和醋酸纤维膜区带电泳、原位底物染色法检测HKs总活性及同工酶活性，用Trander氧化法检测细胞对葡萄糖的净利用。结果SV40MES13细胞株和原代培养的鼠系膜细胞具有相似的HK活性，且主要表达HKⅡ同工酶。PKC激活剂-佛波酯（PMA、PDD）以时间和剂量依赖形式特异地诱导了系膜细胞HKs的活性，并增加了细胞对葡萄糖的净利用。结论系膜细胞主要表达HKⅡ，提示胰岛素参与了系膜细胞葡萄糖的代谢过程；PKC使HK活性上调，增加了细胞对葡萄糖的净利用，提示细胞内葡萄糖流量与HKs活性呈正相关。这将为进一步探讨糖尿病肾病的发病机制提供依据。
Objective To investigate the relationship between diabetic related factors - protein kinase C (PKC) specific regulation of hexokinases (HKs) and cell net glucose utilization, and explore the role of HKs in diabetic nephropathy. Methods SV40 transgenic mouse glomerular mesangial cell line (SV40MES13) was used as a cell model, and glucose-6-phosphate dehydrogenase coupled colorimetry and acetate fiber membrane electrophoresis were used respectively. The in situ substrate staining was used to detect the total activity of HKs And isozyme activity, the use of Trander oxidation of cells to detect the net utilization of glucose. Results SV40MES13 cells and primary cultured mouse mesangial cells had similar activity of HK, and mainly expressed HKⅡ isoenzyme. PKC activator-phorbol ester (PMA, PDD) specifically induced the mesangial cell HKs activity in a time-and dose-dependent manner and increased the cell’s net utilization of glucose. Conclusions Mesangial cells mainly express HKⅡ, suggesting that insulin is involved in the metabolism of glucose in mesangial cells. PKC up-regulates HK activity and increases the net utilization of glucose by cells, suggesting a positive correlation between intracellular glucose flux and HKs activity. This will provide the basis for further exploring the pathogenesis of diabetic nephropathy.