高糖通过调节腺苷酸活化蛋白激酶/聚二磷酸腺苷核糖聚合酶-1的磷酸化抑制B细胞淋巴瘤/白血病-6基因表达的研究

Study on high glucose inhibitting the expression of BCL-6 through AMPKα/PARP-1 phosphorylation

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 inhibiting effect of high concentration glucose on the expression of BCL-6 through AMPKα/PARP-1 phosphorylation pathway in endothelial cells （ECs）. Methods ECs isolated from human umbilical vein endothelial cells （HUVECs）were incubated with high concentration glucose for 24 hours. The phosphorylation of AMPKα Thr-172 and PARP1 Ser-177 in ECs were detected by RT-PCR and western blot. After 4 h pretreatment with AICAR （1 mmol/L）, ECs were co-cultured with high glucose （30 mmol/L） for 24 h. The phosphorylation of AMPKα Thr-172 and PARP1 Ser-177 and the expression of BCL-6 were determined by PR-PCR, western blot and immunofluorescence. Results High glucose decreased the expression of BCL-6 and increased the expression of VCAM-1, MCP- 1 and MCP-3. High glucose dose-dependently attenuated the phosphorylation of AMPKα Thr-172 and PARP1 Ser-177 and the expression of BCL-6 in ECs,which could be reversed by AICAR. Conclusion High concentrations of glucose can reduce the expression of BCL-6 by inhibiting AMPARP-1 phosphorylation arid enhance the effect of inflammation.