补体旁路激活产物刺激内皮细胞NF-κB、p38MAPK、JAK2通路活化及抑制剂的干预研究

Activation of NF-κB, p38MAPK, and JAK2 in Endothelial Cells Induced by Activated Complement Alternative Pathway and Intervention by Inhibitors

研究补体旁路激活产物刺激人微血管内皮细胞NF-κB、p38MAPK、JAK2通路活化的作用及相关抑制剂对其活化的干预作用。采用眼镜蛇毒因子(cobra venom factor,CVF)特异激活血清补体旁路。将旁路活化的血清作用于人微血管内皮细胞,通过ELISA测定NF-κB、p38MAPK及JAK2的活化情况,分别采用上述3个通路的抑制剂PDTC、SB203580、AG490干预其活化及黏附分子ICAM-1的表达。结果显示,补体旁路激活产物能导致人微血管内皮细胞JAK2、p38MAPK、NF-κB通路活化,分别在刺激2,15,30 min后,分别检测到3个通路的活化高峰;PDTC、SB203580、AG490可分别抑制NF-κB、p38MAPK、JAK2的活化。AG490对内皮细胞活化后的ICAM-1蛋白表达有明显的抑制作用,PDTC、SB203580对ICAM-1表达没有明显影响。表明AG490对JAK2通路的抑制可有效下调内皮细胞ICAM-1的表达,提示JAK2通路有可能是补体相关炎症的潜在干预靶点。

The activation of NF-κB, p38MAPK, and JAK2 in human microvascular endothelial cells induced by activated complement alternative pathway and intervention by inhibitors were investigated in this paper. Cobra venom factor （CVF） was used to activate normal human serum. The activation of NF-κB, p38MAPK, and JAK2 were measured by ELISA method after exposure of human microvascular endothelial cells to activated complement of the alternative pathway. The effect of PDTC, SB203580, and AG490 on inhibiting activation of NF-κB, p38MAPK and JAK2, and the expression of ICAM-1 were measured after pretreated with endothelial cells. The results showed that JAK2, p38MAPK, and NF-κB were activated after exposure of human microvascular endothelialcells to activated complement. The maximum activation of JAK2, p38MAPK, and NF-κB was arrived after cells exposed to activated serum complement for 2, 15, 30 min, respectively. The activation of JAK2, p38MAPK, and NF- κB was inhibited by AG490, SB203580, and PDTC, respectively. AG490 significantly inhibited the expression of ICAM-1 in endothelial cells induced by activated complement alternative pathway, whereas PDTC and SB203580 had no effect. Inhibition of JAK2 by AG490 effectively down-regulated the expression of ICAM- 1, indicates that JAK2 may be a potential target for intervention in inflammation induced by complement.