天香丹对动脉粥样硬化秽浊痰阻证ApoE^(-/-)小鼠血清炎性细胞因子及NF-κB p65信号通路的影响

Effect of Tianxiangdan on Inflammatory Factor and NF-κB p65 Signal Pathway of Atherosclerosis ApoE^(-/-) Mice with Phlegm Turbidity Obstruction Syndrome

目的观察天香丹对动脉粥样硬化(atherosclerosis,AS)秽浊痰阻证载脂蛋白E基因敲除[apolipoprotein E gene knock-out,ApoE^((-/-))]小鼠血清中炎性细胞因子白细胞介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)以及主动脉组织中核因子-κB p65(NF-κB p65)信号通路的影响,探讨天香丹防治AS的可能机制。方法 48只8周龄ApoE^((-/-))小鼠分为正常对照组(12只,给予普通饲料室温饲养),高脂饲料复合气候箱干预组(36只)。喂养12周后,成功复制ApoE^((-/-))小鼠AS秽浊痰阻证模型后,分为3组:模型组、天香丹组、阿托伐他汀组。12周后处死各组小鼠,采用酶联免疫双抗夹心(ELISA)法检测血清中IL-1β和TNF-α含量;蛋白免疫印迹(Western blot)法检测主动脉组织中NF-κB p65蛋白表达水平。结果模型组IL-1β、TNF-α浓度高于正常对照组(P<0.01),天香丹组、阿托伐他汀组IL-1β、TNF-α浓度低于模型组(P<0.01);天香丹组与阿托伐他汀组比较,差异均无统计学意义(P>0.05);此外,模型组主动脉组织中NF-κB p65蛋白磷酸化水平高于正常对照组(P<0.01),天香丹组、阿托伐他汀组NF-κB p65蛋白磷酸化水平均低于模型组(P<0.01);天香丹组与阿托伐他汀组比较,差异无统计学意义(P>0.05)。结论天香丹能够防治AS,其机制可能与抑制NF-κB p65信号通路的活性,下调炎性细胞因子IL-1β、TNF-α表达水平,进而抑制炎症反应有关。

Objective To observe the effect of Tianxiangdan （TXD） on expressions of IL-1β, tumor necrosis factor-α （TNF-α）, and nuclear factor-κB （NF-κB） p65 in apolipoprotein E gene knock-out [ApoE（-/-）] atherosclerosis （AS） mice with phlegm turbidity obstruction syndrome （PTOS）, and to ex- plore the possible mechanism of TXD for prevention and treatment of AS. Methods A total of 48 ApoE（-/-）mice （8 weeks old） were divided into the normal control group （A, n =12） and the intervention group （B, n =36）. Mice in the normal control group were fed with common diet at room temperature, while those in Group B were intervened in artificial complex climate box. After 12 weeks feeding, AS mod- el with PTOS were successfully duplicated in mice. Then they were divided into three groups, the model group, the TXD group, and the Atorvastatin group. Mice in each group were scarified 12 weeks later. Ser- um levels of IL-1β and TNF-αin each group were detected. Furthermore, the expression level of NF-κB p65 in aortic tissue was detected by Western blot. Results Levels of serum IL-1β and TNF-α in the mod-el group were significantly higher than those in the normal control group （P 〈0.01 ）. Compared with the model group, serum levels of IL-1β and TNF-α were reduced obviously in the TXD group and the Atorvas- tatin group （P 〈0.01 ）. No statistical difference in serum levels of IL-1 β and TNF-α existed between the TXD group and the Atorvastatin group （P 〉0.05）. Furthermore, phosphorylation level of NF-κB p65 pro- tein in aortic tissue was higher in the model group than in the normal control group （P 〈0.01 ）. The phos- phorylation level of NF-κB p65 protein was lower in the Atorvastatin group and the TXD group than in the model group （P 〈0.01 ）. No statistical difference in the phosphorylation level of NF-κB p65 protein existed between the TXD group and the Atorvastatin group （P 〉0.05）. Conclusion The mechanism of TXD in the prevention and treatment of AS might be associated with suppressing the activation of NF-κB p65 sig- naling pathways, and down-regulating expression levels of inflammatory cytokines IL-1 β and TNF-α, thus further inhibiting inflammatory reactions.