摘要
目的研究过氧化物酶体增殖物激活受体(Peroxisome Proliferators-activated Receptor,PPAR)-β/δ及其激动剂GW0742、抑制剂GSK0660在RAW264.7泡沫细胞模型中的作用。方法本研究选用RAW264.7细胞,共分为6组,分别是对照组、LPS组、ox-LDL组、模型组、激动剂组和抑制剂组。实时定量PCR(Real-time Quantitative PCR)用来检测PPARβ/δ和单核细胞趋化蛋白(monocyte chemoattractant protein,MCP)-1 m RNA的水平,蛋白质印迹法(Western Blotting,WB)检测PPARβ/δ和MCP-1蛋白的表达,酶联免疫吸附试验(Enzyme-linked Immunosorbent Assay,ELISA)测定细胞培养液上清中炎症因子肿瘤坏死因子(tumor necrosis factor,TNF)-α,白介素(interleukin,IL)-10和MCP-1的水平。结果对照组、LPS组、ox-LDL组、模型组、激动剂组的PPARβ/δ蛋白质和m RNA水平逐渐增加(P<0.01),而抑制剂组PPARβ/δ蛋白质和m RNA水平则明显低于模型组和激动剂组(P<0.01)。MCP-1蛋白质和m RNA的水平在对照组、ox-LDL组、LPS组、模型组逐渐增加(P<0.01),而激动剂组明显降低(P<0.01),抑制剂组又有所增加(P<0.01)。总之,MCP-1的蛋白质和m RNA水平在各组之间差异显著(P<0.01)。细胞培养液中TNF-α、MCP-1的水平LPS组、ox-LDL组、模型组明显升高(P<0.01),而激动剂组明显降低(P<0.01),抑制剂组又有所增加(P<0.01)。而IL-10在激动剂组和抑制剂组则表现出相反的趋势。结论 GW0742可以显著增加PPARβ/δ的表达,降低炎症因子MCP-1、TNF-α的水平,升高IL-10的水平;GSK0660则发挥完全相反的作用。
Objective We aimed to examine the role of peroxisome proliferator-activated receptor（PPAR）-β/δ and its ligands GW0742 and GSK0660 in RAW264.7 foam cells. Methods Real-time quantitative PCR analysis was used to detect PPARβ/δ and monocyte chemoattractant protein（MCP）-1 m RNA, western blot analysis（WB） was used to measure levels of PPARβ/δ and MCP-1 protein. Levels of tumor necrosis factor（TNF）-α, interleukin（IL）-10 and MCP-1 in cell culture supernatant were measured by enzyme-linked immunosorbent assay（ELISA）. Results PPARβ/δ protein and m RNA were gradually increased in the LPS group, ox-LDL group, model group and GW0742 group（P〈0.01）, and that of the GSK0660 group were significantly lower than the model group and GW0742 group（P〈0.01）. MCP-1protein and m RNA were significantly increased in the LPS group, ox-LDL group and model group（P〈0.01）, and decreased in the GW0742 group（P〈0.01）, while in the GSK0660 group MCP-1protein and m RNA were significantly increased and higher than the model group（P〈0.01）. Moreover, levels of TNF-α and MCP-1 in cell culture supernatant were risen markedly in the LPS group, ox-LDL group and model group（P〈0.01）, decreased in the GW0742 group（P〈0.01）, and risen again in the GSK0660 group（P〈0.01）. While IL-10 level in cell culture supernatant were risen significantly in the LPS group, ox-LDL group, model group and GW0742 group（P〈0.01）, and decreased in the GSK0660 group（P〈0.01）. Conclusions GW0742 significantly increased the expression of PPARβ/δ, reduced TNF-α, MCP-1 levels, and increased the level of IL-10. However, GSK0660 played a totally opposite role.
出处
《中国分子心脏病学杂志》
CAS
2014年第6期1139-1143,共5页
Molecular Cardiology of China
基金
国家自然科学基金重大研究计划项目(91339101)
