宁心痛颗粒含药血清对THP-1源性巨噬细胞TLR4、Syk、ERK、Paxillin蛋白磷酸化水平与细胞吞噬功能的影响  

Effect of Ningxintong Granule- containing Serum on the Phosphorylated Level of TLR4,Syk,ERK,Paxillin and Phagocytic Function of THP- 1- MΦ

作　　者：陈蕊[1] 高颖[2,3] 顾宁[4] 

机构地区：[1]南京中医药大学第一临床医学院,江苏南京210023 [2]南京中医药大学博士后流动站,江苏南京210023 [3]第二军医大学附属公利医院,上海200135 [4]南京中医药大学第三附属医院,江苏南京210001

出　　处：《时珍国医国药》2016年第4期784-787,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金面上项目(No.81173399)

摘　　要：目的观察宁心痛颗粒含药血清对"轻度氧化修饰低密度脂蛋白-Toll样受体4-巨噬细胞"途径的干预作用,探讨其稳定动脉粥样硬化(Atherosclerosis,AS)斑块、抑制AS进展的分子机制。方法将THP-1源性巨噬细胞分为空白组、模型组、空白血清组、宁心痛颗粒含药血清组4组。采用免疫印迹法检测脾酪氨酸激酶(spleen tyrosine kinase,Syk)、细胞外信号调节激酶(extracellular regulated protein kinases,ERK1/2)、桩蛋白(Paxillin)磷酸化水平随时间变化情况,以及宁心痛颗粒含药血清对Syk、ERK、Paxillin蛋白磷酸化水平的影响;采用免疫沉淀联合免疫印迹法检测Toll样受体4(Toll like receptor 4,TLR4)蛋白磷酸化水平随时间变化情况,以及宁心痛颗粒含药血清对TLR4蛋白磷酸化水平的影响;采用中性红吞噬实验检测巨噬细胞吞噬功能改变。结果 TLR4、Syk、ERK、Paxillin分别于15,10,15,30min达到磷酸化水平高峰;与空白组相比,模型组及空白血清组TLR4、Syk、ERK、Paxillin磷酸化水平升高、细胞吞噬功能增强(P<0.05);与模型组和空白血清组相比,含药血清组TLR4、Syk、ERK、Paxillin磷酸化水平降低、细胞吞噬功能减弱(P<0.05),但仍高于空白组。结论宁心痛颗粒含药血清可能通过"mm LDL-TLR4-巨噬细胞"途径影响巨噬细胞吞噬功能从而具有稳定AS斑块、干预AS作用。Objective To observe the effect of Ningxintong granule- containing serum on ＂ mm LDL- TLR4- MΦ＂,and to explore its possible anti- atherosclerosis mechanism. Methods THP- 1- MΦ cells were divided into 4 groups： blank control group,model group,blank serum group,and Ningxintong granule- containing serum group（ drug- containing serum group）.Western- blot was used to detect the time phase of p- Syk,p- ERK,p- Paxillin,and the influence of drug- containing serum on the degree of p- Syk,p- ERK,p- Paxillin. Western- blot ＋ IP were used to detect the time phase of p- TLR4,and the influence of drug- containing serum on the degree of p- TLR4. Results TLR4,Syk,ERK and Paxillin reached the peak of phosphorylation at 15 min,10min,15 min,30min respectively. Compared with blank control group,the level ofp- TLR4,p- Syk,p-ERK and p- Paxillin was higher,and MΦ ＇s phagocytic function stronger in model group and blank serum group（ P〈0. 05）.Compared with model group and blank serum group,the level of p- TLR4,p- Syk,p- ERK and p- Paxillin was lower,and MΦ＇s phagocytic function weaker in drug- containing serum group（ P〈0. 05）. Conclusion Ningxintong granule- containing serum can inhibit phagocytic function and have the anti- atherosclerotic effects through ＂ mm LDL- TLR4- MΦ＂ pathway.

关 键 词：宁心痛颗粒 含药血清 轻度氧化修饰低密度脂蛋白 Toll样受体4 脾酪氨酸激酶 巨噬细胞 动脉粥样硬化 

分 类 号：R285.5[医药卫生—中药学]