当归补血汤对氧化低密度脂蛋白激活RAW264.7细胞中核转录因子-κBp65 mRNA表达的影响

Effect of Danggui Buxue Decoction on Nuclear Factor Kappa B p65 mRNA Expression in RAW264.7 Cells Stimulated by Oxidized Low-density Lipoprotein

【目的】观察当归补血汤含药血清对氧化低密度脂蛋白(Ox-LDL)激活小鼠单核细胞系RAW264.7细胞中核转录因子-κBp65(NF-κBp65)mRNA表达的影响。【方法】将新西兰兔8只随机分为空白血清组和含药血清组,每组各4只,分别灌胃生理盐水和当归补血汤水煎液(剂量为8.4g·kg-1·d-1),分离血清。将RAW264.7细胞悬液接种于培养瓶中,培养24h,分为空白对照组、Ox-LDL组、PAR组(终浓度为10μmol/L的小菊白内酯)、体积分数10%的含药血清组。除空白对照组外,其他各组分别加入100μg/mLOx-LDL刺激4h,收集细胞。采用实时荧光定量PCR法检测各组细胞NF-κBp65 mRNA的表达量。【结果】Ox-LDL组与空白对照组比较,NF-κBp65 mRNA的表达量增加(P<0.05);含药血清组与Ox-LDL组比较,NF-κBp65 mRNA的表达量减少(P<0.05)。【结论】当归补血汤含药血清能下调NF-κBp65 mRNA的表达量,抑制、阻断NF-κBp65的表达可能是当归补血汤抗动脉粥样硬化的作用机制之一。

Objective To investigate the influence of serum containing Danggui Buxue Decoction （DBD） on transcription factor of nuclear factor kappa B （NF-KB） activity stimulated by oxidized low-density lipoprotein （Ox-LDL） in RAW264. 7 cells, and to explore the cell signal transduction mechanism of DBD in counteracting atherosclerosis （AS）. Methods Eight new Zealand rabbits were randomized into 2 groups： blank serum group was given gastric gavage of normal saline, and drug-containing serum group was given DBD （8.4 g · kg^-1 · d^-1 ）. After treatment, the rabbits were executed and the serum was separated. RAW264.7 cells were incubated in the culture flask for 24 hours, and then were divided into blank control group, Ox-LDL group, inhibitor group （parthenolide at the concentration of 10 μmol/L）, and drug-containing serum group （including 10% volume fraction of serum containing DBD）. Except the blank control group, the other groups were stimulated by 100 μg/ mL Ox-LDL for 4 hours, and then RAW264.7 cells were collected. NF-κBp65 mRNA expression in RAW264.7 cells was detected with real-time fluorescent quantitative polymerase chain reaction （PCR）. Results Ox-LDL group had higher expression of NF-κBp65 mRNA than the blank control group （ P 〈 0.05 ）, and the expression of NF-κBp65 mRNA was reduced in drug-containing serum group as compared with that in Ox-LDL group （P 〈 0. 05）. Conclusion Serum containing DBD can down-regulate NF-κBp65 mRNA expression in RAW264.7 cells, and inhibition and blockage of NF-κB p65 mRNA expression may be one of the anti-AS mechanisms of DBD.