Wnt/β-catenin信号通路在可溶性Klotho蛋白抑制THP-1泡沫细胞形成过程中的作用

Effect of Wnt/β-catenin signaling pathway in process of soluble Klotho protein suppressing THP-1 foam cell

目的探讨Wnt/β-catenin信号通路在可溶性Klotho蛋白抑制THP-1泡沫细胞形成过程中的作用。方法THP-1单核细胞在佛波酯的作用下诱导分化成为THP-1巨噬细胞,再由氧化型低密度脂蛋白诱导其转变为泡沫细胞,可溶性Klotho蛋白和β-甘油磷酸进行预处理。采用油红O染色法观察各组细胞内脂滴的形成情况,酶荧光法检测细胞内总胆固醇(TC)、游离胆固醇(FC)及胆固醇酯(CE)水平,Western blot法检测β-catenin、c-myc与cyclinD1蛋白表达,免疫荧光染色法检测细胞内β-catenin表达及分布情况。结果与阳性对照组比较,Klotho蛋白组TC、CE的水平均明显降低,β-catenin、c-myc与cyclinD1蛋白表达明显减少,β-catenin表达和分布明显减少;β-甘油磷酸组TC、CE水平明显升高,β-catenin、c-myc与cyclinD1蛋白表达明显增加,β-catenin表达和分布明显增加,差异均有统计学意义(P<0.05)。与β-甘油磷酸组比较,Klotho蛋白+β-甘油磷酸组TC、CE水平明显降低,β-catenin、c-myc与cyclinD1蛋白表达明显减少,β-catenin表达和分布明显减少,差异有统计学意义(P<0.05)。结论可溶性Klotho蛋白可能通过抑制Wnt/β-Catenin信号通路从而抑制THP-1泡沫细胞形成。

Objective To investigate the role of Wnt/β-catenin signaling pathway in the process of soluble Klotho protein inhibiting THP-1 foam cell.Methods THP-1 monocytes were induced and differentiated into the THP-1 macrophages under the action of phorbol ester,then induced by oxidized low density lipoprotein to transform into the foam cells,the pretreatment was performed by soluble Klotho protein andβ-glycerophosphoric acid.The intracellular lipid droplets formation was observed by oil red O staining.The levels of intracellular total cholesteml(TC),free cholesterol(FC)and cholesterol ester(CE)were detected by the enzyme fluorescence method.The expressions ofβ-catenin,c-myc and cyclinD1 protein were determined by Western blot.The expression levels and distribution ofβ-catenin were observed by the immunofluorescence staining.Results Compared with the positive control group,the TC and CE levels in the Klotho protein group were significantly decreased,β-catenin,c-myc and cyclinD1 proteins expressions were significantly decreased,the expression and distribution ofβ-catenin were significantly decreased;the TC and CE levels in theβ-glycerophosphoric acid group increased,β-catenin,c-myc and cyclinD1 protein expressions were significantly increased,and the differences were statistically significant(P<0.05).Compared with theβ-glycerophosphoric acid group,the levels of TC and CE in the Klotho protein+β-glycerophosphoric acid group were significantly decreased,the expressions ofβ-catenin,c-myc and cyclinD1 protein expressions were significantly decreased,the expression and distribution ofβ-catenin were significantly decreased,and the differences were statistically significant(P<0.05).Conclusion The soluble Klotho protein inhibits the THP-1 foam cell formation possibly by suppressing the Wnt/β-catenin signal pathway.