黄连解毒汤含药血清激活PPARγ诱导RAW264.7源性泡沫细胞向M2表型极化

Activation of PPARγ by Huanglianjiedu Decoction serum induced RAW264.7-derived foam cells to polarization of M2 phenotype

目的:通过体外细胞实验观察黄连解毒汤干预RAW264.7源性泡沫细胞PPARγ表达对细胞极化的影响,探讨黄连解毒汤抗动脉粥样硬化的部分作用机制。方法:氧化低密度脂蛋白(ox-LDL)诱导RAW264.7源性泡沫细胞;油红O染色法鉴定细胞泡沫化;以一定浓度的黄连解毒汤含药血清干预细胞,流式细胞术鉴定M2型巨噬细胞表达;酶联免疫吸附测定(ELISA)检测细胞上清IL-1β、IL-10、转化生长因子-β(TGF-β)和肿瘤坏死因子-α(TNF-α)的含量;蛋白免疫印迹法(Western blot)测定细胞过氧化物酶体增殖剂激活受体γ(PPARγ),精氨酸酶1(Arg-1)的蛋白表达水平。结果:油红O染色显示,泡沫细胞模型制备成功。含药血清干预后,与模型组相比,巨噬细胞CD206分子表达阳性数量明显升高(P<0.01)。细胞上清IL-1β、TNF-α的表达下调,IL-10、TGF-β的表达明显增多(P<0.05,P<0.01),PPARγ、Arg-1的蛋白表达水平明显升高(P<0.05,P<0.01)。结论:黄连解毒汤可通过激活PPARγ促进ox-LDL诱导的RAW264.7源性泡沫细胞向M2表型极化,发挥抗炎作用,这可能是其防治动脉粥样硬化机制之一。

Objective:To observe the effect of Huanglianjiedu decoction on the polarization of RAW264.7-derived foam cell PPARγ,and to explore the mechanism of Huanglianjiedu decoction against atherosclerosis.Methods:Oxidized low density lipoprotein(ox-LDL)induced RAW264.7 foam cells was identified by Oil red O staining.The expression of M2-type macrophages was determined by flow cytometry after intervention with certain concentration of serum containing Huanglianjiedu decoction.Enzyme linked immunosorbent assay(ELISA)was used to detect the contents of IL-1β,IL-10,TGF-βand TNF-αon cells.Western blot was performed to determine the protein expression levels of peroxisome proliferator activated receptorγ(PPARγ)and Arginidase 1(Arg-1).Results:Oil red O staining showed that the foam cell model was successfully prepared.After intervention,the positive expression of CD206 molecules in macrophages was significantly increased compared with that in the model group(P<0.01).The expressions of IL-1βand TNF-αin the supernatant of cells were down-regulated,the expressions of IL-10 and TGF-βwere significantly increased(P<0.05,P<0.01),and the protein expressions of PPARγand Arg-1 were significantly increased(P<0.05,P<0.01).Conclusion:Huanglianjiedu decoction can promote ox-LDL-induced RAW264.7-derived foam cells to the M2-phenotype polarization by activating PPARγ.It may play an anti-inflammatory role,which may be one of the mechanisms for its prevention and treatment of atherosclerosis.