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目的探讨高浓度葡萄糖对B细胞淋巴瘤/白血病-6基因(BCL-6)的影响及其是否通过腺苷酸活化蛋白激酶α/聚二磷酸腺苷核糖聚合酶-1(AMPKα/PARP 1)磷酸化途径。方法分离培养人脐静脉内皮细胞,用不同浓度的葡萄糖孵育脐静脉内皮细胞24 h,RT-PCR和Western blot检测检测内皮细胞AMPK hr172和PARP-1 Ser-177的磷酸化、BCL-6及炎症因子血管细胞黏附分子-1(VCAM-1)、单核细胞去化因子-1(MCP-1)和MCP-3的表达。用不同浓度的AMPK磷酸化激动剂AICAR(0、0.5、1mmol/L)孵育脐静脉内皮细胞4 h;Western blot检测内皮细胞AMPKhr 172和PARP-1 Ser-177的磷酸化及BCL-6表达。用AICAR预处理内皮细胞4 h,然后加入高浓度葡萄糖(30 mnol/L)作用于内皮细胞24 h,Western blot及免疫荧光检测内皮细胞AMPK hr 172和PARP-1 Ser-177的磷酸化及BCL-6表达。结果高浓度葡萄糖可抑制AMPK hr 172和PARP-1 Ser-177的磷酸化,降低内皮细胞BCL 6的表达,同时,炎症因子VCAM-1、MCP-1和MCP-3表达增加。AICAR能增强内皮细胞AMPKhr 172和PARP 1Ser-177的磷酸化,BCL-6表达增加。AICAR可逆转高浓度葡萄糖对AMPKhr 172和PARP 1 Ser-77的磷酸化及BCL-6表达的抑制作用。结论高浓度葡萄糖通过抑制AMPARP-1磷酸化减少BCL-6的表达,从而使炎症表达增加。
Objective To investigate the effect of high concentration glucose on B cell lymphoma / leukemia-6 gene (BCL-6) and whether it is mediated by AMPKα / PARP 1, Phosphorylation pathway. Methods Human umbilical vein endothelial cells were isolated and cultured. Human umbilical vein endothelial cells were incubated with different concentrations of glucose for 24 h. The phosphorylation of AMPK hr172 and PARP-1 Ser-177, BCL-6 and inflammation were detected by RT-PCR and Western blot The expression of vascular endothelial cell adhesion molecule-1 (VCAM-1), monocyte chelator-1 (MCP-1) and MCP-3. Umbilical vein endothelial cells were incubated with different concentrations of AMPK phosphorylation agonist AICAR (0, 0.5, 1 mmol / L) for 4 h. The phosphorylation and BCL-6 expression of AMPKhr 172 and PARP-1 Ser-177 in endothelial cells were detected by Western blot. Endothelial cells were pretreated with AICAR for 4 h and then treated with high concentration of glucose (30 mnol / L) for 24 h. The phosphorylation of AMPK hr 172 and PARP-1 Ser-177 and BCL -6 expression. Results High concentrations of glucose inhibited the phosphorylation of AMPK hr 172 and PARP-1 Ser-177 and decreased the expression of BCL 6 in endothelial cells. Meanwhile, the expression of inflammatory factors such as VCAM-1, MCP-1 and MCP-3 increased. AICAR enhanced the phosphorylation of endothelial cells AMPKhr 172 and PARP 1Ser-177, and increased BCL-6 expression. AICAR reverses the inhibitory effect of high concentration glucose on the phosphorylation of AMPKhr 172 and PARP 1 Ser-77 and on the expression of BCL-6. Conclusion High concentration of glucose can reduce the expression of BCL-6 by inhibiting the phosphorylation of AMPARP-1, so that the expression of inflammation is increased.