蒲黄总黄酮含药血清对巨噬细胞自体吞噬及Akt/mTOR信号通路的影响  

Effect of total flavonoid-containing serum of Pollen Typhae on autophagy of macrophages and its effect on Akt/mTOR signaling pathway

作　　者：王丹[1] 孙刚[1] 王瑶[1] 

机构地区：[1]贵阳中医学院第一附属医院心内科,550002

出　　处：《实用医学杂志》2017年第16期2653-2657,共5页The Journal of Practical Medicine

基　　金：贵州省科学技术基金项目(编号:黔科合LH字[2014]7350号)

摘　　要：目的研究蒲黄总黄酮(TYTF)含药血清对巨噬细胞自体吞噬的作用及其对Akt/mTOR信号通路的影响。方法采用高、中、低剂量TYTF水溶液灌胃干预SD大鼠,制备含药血清,采用MMT法观察含药血清对细胞增殖的影响,选择浓度为20%的含药血清与巨噬细胞共培养,透射电镜下观察巨噬细胞自噬体的变化,检测白介素-10(IL-10)和干扰素γ(IFN-γ)及自噬相关基因Akt和mTOR mRNA和蛋白的表达。结果 TYTF含药血清在10%~40%的浓度范围内的OD值与正常对照组的OD值比较,差异无统计学意义(P>0.05)。在透射电镜下观察,TYTF各剂量组中的自噬体的表达均明显增加,而正常对照组自噬体表达不典型。TYTF各剂量组含药血清Beclin1蛋白表达均显著高于正常对照组,差异具有统计学意义(P<0.05),而p-Akt及p-mTOR蛋白表达显著低于正常对照组(P<0.05),Akt及mTOR蛋白表达与正常对照组比较差异无统计学意义(P>0.05)。TYTF各剂量组含药血清Beclin1 mRNA表达均显著高于正常对照组(P<0.05),而Akt及mTOR mRNA表达显著低于正常对照组(P<0.05)。TYTF各剂量组含药血清IL-10的表达显著高于正常对照组(P<0.05),而IFN-γ的表达显著低于正常对照组(P<0.05)。结论 TYTF含药血清具有抗动脉粥样硬化的作用,可能抑制Akt/mTOR信号通路的激活,从而诱导巨噬细胞的自噬活动,减少斑块内巨噬细胞的浸润,并通过抑制炎症反应因子的分泌,抑制炎症反应活动,进而达到促进动脉粥样硬化易损斑块稳定性的目的 。Objective To study the effect of total flavonoids of Pollen Typhae on the autophagy of macrophages and its effect on Akt/mTOR signaling pathway.Methods To observe the effect of serum containing drugs on the proliferation of SD rats.The serum containing 20% of the drug-containing serum and the macrophages were selected by the method of MMT.Interleukin（IL-10）,interferon-γ（IFN-γ）and autophagy-related genes Akt and mTOR mRNA and protein were detected.ResultsThere was no significant difference between the OD value of TYTF serum in the concentration range of 10%~40% and so as the OD value of normal control（P〈0.05）.The expression of autophagosomes in each dose group of TYTF was significantly increased under TEM.The autophagic expression was not typical in the normal control group.The expression of Beclin1 protein in TYTF group was significantly higher than that in normal control group（P〈0.05）,while the expression of p-Akt and p-m TOR protein was significantly lower than that of normal control group（P〈0.05）.Akt and m TOR protein expression compared with the normal control group was not significantly different（P〈0.05）.The expression of Beclin1 mRNA in TYTF group was significantly higher than that in normal control group（P〈0.05）,while the expression of Akt and mTOR mRNA was significantly lower than that of normal control group（P〈0.05）.The expression of IL-10 in TYTF group was significantly higher than that in normal control group（P〈0.05）.The expression of IFN-γwas significantly lower than that of normal control group（P〈0.05）.Conclusions The total flavonoid serum has antiatherosclerotic effect.The mechanism may be related to inhibition of the activation of Akt/mTOR signaling pathway,including macrophage autophagy activity induction,plaque macrophage infiltration reduction and inflammatory response factors secretion inhibition,inflammatory response activities inhibition.Thereby it can promote atherosclerosis plaque stability of the purpose of vulnerability.

关 键 词：蒲黄总黄酮 巨噬细胞 AKT/MTOR 动脉粥样硬化 

分 类 号：R285.5[医药卫生—中药学]