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目的:通过鸟苷酸结合蛋白(G-蛋白)对不规则趋化因子fractalkine(FKN)诱导单个核细胞合成核因子κB(NF-κB)、肿瘤坏死因子-α(TNF-α)的影响,探讨FKN及其受体CX3CR1可能存在的信号转导机制及卡托普利的干预作用。方法:将抗凝血用Ficoll密度梯度离心法分离外周血单个核细胞,随机分为空白对照组、FKN组、PTX组、RO31-8220组、PDTC组及卡托普利组,应用免疫组化检测各组单个核细胞中NF-κB表达情况,应用酶联免疫法检测各组培养液中TNF-α的表达水平。结果:FKN组与空白组比较NF-κB、TNF-α表达明显增多(P<0.05);G-蛋白阻断剂PTX组与FKN组比较NF-κB、TNF-α表达明显减少(P<0.05);PKC阻断剂RO31-8220组与FKN组比较NF-κB、TNF-α表达减少(P<0.05);NF-κB抑制剂PDTC组TNF-α表达较FKN组明显减少(P<0.05);卡托普利组较FKN组NF-κB、TNF-α表达部分减少(P<0.05)。结论:FKN/CX3CR1有增加单个核细胞表达NF-κB、TNF-α的作用;而卡托普利可通过减弱FKN/CX3CR1诱导单个核细胞合成NF-κB、TNF-α,抑制炎症反应和抗动脉粥样硬化;FKN/CX3CR1影响动脉粥样硬化的信号转导机制可能是通过以下级联反应实现的:FKN+CX3CR1→G蛋白→PKC→NF-κB→TNF-α。
OBJECTIVE: To investigate the effects of guanylate binding protein (G-protein) on nuclear factor κB (NF-κB) and tumor necrosis factor-α (TNF-α) induced by irregular chemokine fractalkine (FKN) To investigate the possible signal transduction mechanism of FKN and its receptor CX3CR1 and the effect of captopril. Methods: Peripheral blood mononuclear cells were isolated by Ficoll density gradient centrifugation and divided randomly into blank control group, FKN group, PTX group, RO31-8220 group, PDTC group and captopril group. Immunohistochemistry The expression of NF-κB in each group of mononuclear cells was detected. The expression of TNF-α in each group was detected by enzyme-linked immunosorbent assay. Results: Compared with blank group, the expression of NF-κB and TNF-α in FKN group was significantly increased (P <0.05); the expression of NF-κB and TNF-α in PTK group was significantly decreased compared with FKN group ); Compared with FKN group, the expression of NF-κB and TNF-α in PKC blocker RO31-8220 group decreased (P <0.05); the expression of TNF-α in NF-κB inhibitor PDTC group decreased significantly compared with FKN group ; Captopril group than FKN group NF-κB, TNF-α part of the reduction (P <0.05). Conclusion: FKN / CX3CR1 can increase the expression of NF-κB and TNF-α in mononuclear cells. Captopril can inhibit the synthesis of NF-κB and TNF-α by inhibiting the expression of FKN / CX3CR1 in mononuclear cells Atherosclerosis; FKN / CX3CR1 affect atherosclerosis signal transduction mechanism may be achieved through the following cascade: FKN + CX3CR1 → G protein → PKC → NF-κB → TNF-α.