3种化痰方剂含药血清超滤组分对THP-1源性巨噬细胞活性及泡沫化的影响

Effect of Ultrafiltrated Components of Three Kinds of Serum Containing Phlegm-resolving Prescriptions on Viability of THP-1 Cell-derived Macrophages and Macrophage Differentiation to Foam Cells

【目的】观察3种化痰方剂(黄连温胆汤、瓜蒌薤白半夏汤、二陈汤)含药血清超滤组分对人白血病单核细胞(THP-1)源性巨噬细胞活性及其泡沫化的影响。【方法】以佛波酯(PMA)诱导THP-1为巨噬细胞,经氧化低密度脂蛋白(ox-LDL)处理建立动脉粥样硬化泡沫细胞模型;制备3种化痰方剂含药血清及空白血清经10 KD超滤器超滤后,作用于泡沫细胞模型。采用四甲基偶氮唑盐(MTT)法及油红染色观察各组对巨噬细胞存活率和泡沫化的影响;通过流式细胞术检测各组对巨噬细胞凋亡率的影响。【结果】THP-1经100 nmol/L PMA诱导24 h后,与50μg/m L ox-LDL共孵育24 h,获得泡沫细胞模型。黄连温胆汤含药血清超滤成分作用于巨噬细胞,可明显提高存活率,抑制泡沫化,降低细胞凋亡率。二陈汤和瓜蒌薤白半夏汤含药血清超滤成分可增加泡沫化,未见显著抗凋亡作用。【结论】黄连温胆汤具有抗动脉粥样硬化作用,可能与其能抗巨噬细胞凋亡,降低巨噬细胞脂质吞噬作用相关;二陈汤和瓜蒌薤白半夏汤可能具有提高巨噬细胞脂质吞噬功能的作用。

Objective To observe the effect of ultrafiltrated components of three kinds of serum containing phlegm-resolving prescriptions （Huanglian Wendan Decoction, Gualou Xiebai Banxia Decoction, and Erchen Decoction, respectively） on viability of human leukemic monocytic cell line THP-1 derived macrophages and macrophage differentiation to foam cells. Methods THP-1 cells were induced to differentiate into macrophages with phorbol-12-myristate-13-acetate （ PMA） . And then the achieved macrophages were used for the establishment of the atherosclerotic foam cell model after co-incubation with oxidized low density lipoprotein （ ox-LDL ） . Three kinds of serum containing phlegm-resolving prescriptions and blank serum were prepared and ultrafiltered by 10 KD ultrafiltrator, and then were co-cultured with the foam cell model, respectively. The viability of macrophages and the macrophage differentiation to foam cells were tested by methyl thiazolyl tetrazolium （ MTT） assay and oil red staining. Apoptotic rate of macrophages was determined by flow cytometry. Results After THP-1 cells were induced by PMA （100 nmol/L） for 20 hours and then were co-incubated with ox-LDL （50 μg/mL） for 24 hours, a typical atherosclerotic foam cell model was established successfully. The ultrafiltrated components of serum containing Huanglian Wendan Decoction had obvious effects on increasing the macrophage survival rate, decreasing macrophage apoptotic rate, and inhibiting macrophage differentiation to foam cells. The ultrafiltrated components of serum containing Gualou Xiebai Banxia Decoction or Erchen Decoction had an effect on enhancing the macrophage differentiation to foam cells but had no anti-apoptosis action. Conclusion Huanglian Wendan Decoction has anti-atherosclerosis action, which may be related to resisting macrophage apoptosis and decreasing macrophagic lipid-endocytosing function. Erchen Decoction and Gualou Xiebai Banxia Decoction are effective on increasing macrophagic lipid-endocytosing function.