脂联素干预TNF-α作用后的巨噬细胞脂联素受体mRNA表达水平及其吞噬脂质变化

The changes of adiponectin receptor mRNA expressions and lipidosome phagocytosis of macrophages after stimulated by adiponectin-intervened TNF-α

目的探讨脂联素干预TNF-α作用后的巨噬细胞表面脂联素受体mRNA表达水平及其吞噬脂质的变化。方法单核巨噬细胞系用佛波酯诱导,使之分化为巨噬细胞;进而采用不同浓度的TNF-α(0、0.1、1、10、100ng/ml)作用24h(以TNF-α0ng/ml为空白对照组),采用RT-PCR技术观察巨噬细胞表面脂联素受体R1、R2mRNA表达水平。选择0、10ng/ml浓度TNF-α作用后的巨噬细胞,用0.2mg/ml低密度脂蛋白混悬生长液作用24h,油红O染色和比色定量检测巨噬细胞吞噬脂质变化;同时TNF-α作用后的巨噬细胞,进一步用人重组脂联素20μg/ml干预24h,再次观察巨噬细胞表达脂联素受体mRNA水平及吞噬脂质变化。结果巨噬细胞用不同浓度的TNF-α作用后,随着TNF-α浓度梯度增加,脂联素受体R1、R2mRNA表达水平逐渐下降(P<0.05)。TNF-α10ng/ml作用后的巨噬细胞吞噬脂质的量明显高于TNF-α空白对照组(P<0.05);同时10ng/mlTNF-α作用后的巨噬细胞,进一步用脂联素干预,巨噬细胞表达脂联素受体R1、R2mRNA水平均较干预前上调(P<0.05),吞噬脂质的量较干预前减少(P<0.05)。结论脂联素及其受体在调节巨噬细胞吞噬脂质过程中产生重要作用,该作用可能成为预防和治疗动脉粥样硬化发生发展的新靶标。

Objective To investigate the effects of TNF-α on adiponectin receptor mRNA expressions and lipid phagocytosis of macrophages,and its further response to the interference effects of adiponectin. Methods THP-1 cells were induced and differentiated into macrophages by phorbol ester.RT-PCR was used to detect the expression of adiponectin receptor mRNA of the macrophages, which were stimulated with different concentrations of TNF-α（0.1,1,10,100 ng/ml） for 24 h, and TNF-α 0 ng/ml was set as the control.Macrophages that stimulated with TNF-α 0 ng/ml and 10 ng/ml were further cultured in growth-promoting media containing 0.2 mg/ml LDL for 24 h, intracellular fat accumulation was observed using oil red O staining and quantified by colorimetry. Meanwhile the macrophages were further interfered with 20 μg/ml adiponectin,and adiponectin receptor mRNA expressions and lipidosome phagocytosis of macrophages were tested. Results The inhibition of adiponectin receptor mRNA expression in the macrophages increased as the concentration of TNF-α was increasing（P0.05）.When the macrophages were stimulated with TNF-α 10 ng/ml,the accumulation of lipid within the cells was obviously increased compared to TNF-α 0 ng/ml（P0.05）.After the macrophages were further interfered with adiponectin,adiponectin receptor mRNA expression was up-regulated and the accumulation of lipid within the cells was decreased（P0.05）. Conclusion Adiponectin and adiponectin receptor play an important role in regulating lipid phagocytosis of macrophages,which may be a new target for the prevention or treatment of atherosclerosis.