黄连解毒汤含药血清对巨噬细胞自噬相关基因表达的影响

Effects of Huang-Lian-Jie-Du-Decotion containing serum on expressions of autophagy related gene in macrophages

目的:考察黄连解毒汤含药血清对RAW264.7巨噬细胞自噬的影响,初步探讨其抗动脉粥样硬化的作用机制。方法:采用大、中、小剂量黄连解毒汤水煎剂灌胃干预SD大鼠,制备含药血清;经MTT法观察含药血清对细胞增殖的影响后,选择各剂量浓度为20%的含药血清干预体外培养的巨噬细胞,采用荧光定量PCR方法检测自噬相关基因Beclin1和m TOR的mRNA表达水平;Western blot检测Beclin1、p-m TOR蛋白的表达变化。结果:与正常对照组血清相比,黄连解毒汤含药血清干预后诱导了Beclin1 mRNA及蛋白的表达,抑制了m TOR mRNA及p-m TOR蛋白的表达。结论:黄连解毒汤含药血清可诱导RAW264.7巨噬细胞自噬相关基因Beclin1的表达、抑制m TOR的表达,这可能是该方抗动脉粥样硬化作用的重要机制之一。

Objective： To investigate the effects of Huang-Lian-Jie-Du-Decotion containing serum for the autophagy of RAM264. 7 macrophages,and to explore its possible anti-atherosclerosis mechanism. Methods： The SD rats were intervened with large,middle and small dose Huang-Lian-Jie-Du-Decotion to prepare containing drug serum. Cell proliferation was tested by MTT assay. The gene and protein expressions of Beclin1,m TOR was assessed by fluorescent quantitative PCR and Western blot method. Results： Compared with the normal serum group,Huang-Lian-Jie-Du-Decotion containing serum enhanced expression of the mRNA and protein level of Beclin1,and decreased the expression of m TOR and p-m TOR（ P〈0. 05 or P〈0. 01）. Conclusion： Huang-Lian-Jie-Du-Decotion containing serum could regulate the expressions of Beclin1 and m TOR. These might be one of the mechanisms for preventing atherosclerosis.