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目的 探讨肝素结合性表皮生长因子 (HB EGF)对系膜细胞己糖激酶 (HK)活性的自分泌调节作用。方法 以SV40MES13系膜细胞株为细胞模型 ,使用 6 磷酸葡萄糖脱氢酶偶联比色法检测HKs活性 ;观察外源性HB EGF、佛波酯 (PMA)条件培养液和EGF受体阻断剂Genistein对系膜细胞HKs活性的影响。结果 外源性HB EGF和PMA条件培养液均以时间和剂量依赖方式诱导系膜细胞HKs活性。在 12h时诱导活性最大 ,分别增加了 6 4.8%± 7.1% (P <0 .0 1)和 6 7.3%± 5 .7% (P <0 .0 1) ;当HB EGF浓度为 10nmol/L和PMA条件培养液浓度为 10 0 %时 ,诱导活性最大 ,分别增加了 5 0 .8%± 5 .0 % (P <0 .0 1)和 5 7.0 %± 7.7% (P <0 .0 1)。HB EGF受体阻断剂 (酪氨酸蛋白激酶阻断剂 )可阻断两者对HKs活性的诱导作用。结论 HB EGF以自分泌 /旁分泌的形式诱导系膜细胞HK活性 ,可能参与了糖尿病肾病的糖代谢异常过程
Objective To investigate the effect of heparin-binding epidermal growth factor (EGF) on the autocrine regulation of hexokinase (HK) activity in mesangial cells. Methods SV40MES13 mesangial cell line was used as the cell model, and the activity of HKs was detected by 6-phosphoglucose dehydrogenase-coupled assay. Exogenous HB EGF, phorbol myristate (PMA) conditioned medium and EGF receptor antagonist Effect of Genistein on HKs Activity of Mesangial Cells. Results Both exogenous HB EGF and PMA conditioned medium induced HKs activity in mesangial cells in a time-and dose-dependent manner. (P <0.01) and 6 7.3% ± 5.7% (P <0.01), respectively, when the concentration of HB EGF was 10 nmol / L (P <0. 01) and 5 7.0% ± 7.7% (P <0. 0), respectively, when the concentration of PMA was 100% 1). HB EGF receptor blockers (tyrosine protein kinase blockers) can block the induction of both HKs activity. Conclusion HB EGF induces the HK activity of mesangial cells in an autocrine / paracrine manner and may be involved in the abnormal glucose metabolism in diabetic nephropathy