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目的通过当归补血汤对氧化低密度脂蛋白(ox-LDL)激活单核细胞核转录因子-κBp65(NF-κBp65)信号转导通路的作用研究,探讨当归补血汤早期阻断动脉粥样硬化病变发展进程的可能性及其机理。方法新西兰大耳白兔随机分为对照血清组、含药血清组,分别灌胃生理盐水和当归补血汤药液制备血清。将RAW264.7细胞悬液接种于培养瓶中,培养24 h,分为空白对照组、ox-LDL组、抑制剂组、含药血清组,除空白对照组外其余各组均加入100 mg/L ox-LDL刺激细胞。孵育4 h,收集细胞。免疫组化法检测NF-κBp65的表达,免疫印迹法检测RAW264.7细胞中NF-κBp65的蛋白活化量。结果ox- LDL组细胞浆和核内可见大量棕黄色阳性颗粒,含药血清组棕黄色阳性颗粒较少;ox-国LDL组蛋白条带灰度比值为1.288 3±0.013 9,含药血清组为0.918 9±0.015 3,与ox-LDL组比较差异有统计学意义(P<0.05)。结论当归补血汤可下调经ox-LDL刺激后RAW264.7细胞中NF-κBp65的蛋白表达及活化,因此阻断NF-κB信号转导通路可能是当归补血汤抗动脉粥样硬化的作用机理之一。
Objective To investigate the effect of Dangguibuxuetang on the activation of nuclear factor-κBp65 (NF-κBp65) signal transduction pathway by oxidized low-density lipoprotein (ox-LDL), to explore the early blocking of the development of atherosclerotic lesions The possibility of the process and its mechanism. Methods New Zealand white rabbits were randomly divided into control serum group and drug-containing serum group. Serum was prepared by intragastric administration of saline and Dangguibuxuetang liquid. The suspension of RAW264.7 cells was inoculated into culture flasks and cultured for 24 h. The cells were divided into blank control group, ox-LDL group, inhibitor group and drug-containing serum group. Except the blank control group, Lox-LDL stimulates cells. Incubate for 4 h and collect the cells. The expression of NF-κBp65 was detected by immunohistochemistry and the activation of NF-κBp65 protein in RAW264.7 cells was detected by Western blotting. RESULTS: A large number of positive brown granules were found in the cytoplasm and nucleus of ox-LDL group, and there were few brown-yellow positive granules in the serum containing ox-LDL group. The gray scale ratio of ox LDL group was 1.288 3 ± 0.013 9, 0.918 9 ± 0.015 3, which was significantly different from that in ox-LDL group (P <0.05). Conclusion Dangguibuxuetang can down-regulate the protein expression and activation of NF-κBp65 in RAW264.7 cells stimulated by ox-LDL, so blockade of NF-κB signal transduction pathway may be the mechanism of anti-atherosclerosis of Dangguibuxuetang one.