TLR4基因敲除对小鼠内脏脂肪组织免疫细胞及脂肪因子的影响

摘要

目的:探讨TLR4基因敲除对小鼠免疫细胞及脂肪因子的影响.方法:取20周龄的雄性野生型C57BL/6小鼠和TLR4-/-小鼠的脾脏和附睾脂肪组织,分离细胞,用流式检测F4/80、CD11b、CD11c、CD3、CD4、CD8分子的表达;qPCR检测附睾脂肪组织内IL-6、HMGB1、TNF-α、脂联素和抵抗素的表达.结果:与野生型C57BL/6小鼠相比,TLR4-/-小鼠脾脏和附睾脂肪组织中M1型(F4/80+CD11b+CD11c+)巨噬细胞比例上升(P<0.05),M2型(F4/80+ CD11b+ CD11c-)巨噬细胞比例下降(P<0.05),这种趋势在附睾脂肪组织中表现更为显著.同时发现附睾脂肪组织中CD4+T细胞比例下降(P<0.05),CD8+T细胞比例上升(P<0.05);IL-6、HMGB1、抵抗素表达升高(P<0.05);TNF-α和脂联素表达降低(P<0.05).结论:TLR4基因敲除可导致内脏脂肪组织脂肪因子和免疫细胞的紊乱.%Objective:To investigate the effects of TLR4 knockout on immune cells and adipokines in mouse visceral adipose tissue.Methods: Cells were isolated from the spleen and epididymal adipose tissue of 20-week-old male wild type mice C57BL/6 and TLR4-/-.The expression of F4/80,CD11b,CD11c,CD3,CD4 and CD8 in these cells were analyzed by flow cytometry.The expression of IL-6,HMGB1,TNF-α,adiponectin and resistin in epididymal adipose tissue were detected by qPCR.Results: Compared with wild type C57BL/6 mice,the percentage of M1 macrophage which marked F4/80+CD11b+CD11c+ in spleen and epididymal adipose tissue of TLR4-/-mice increased(P<0.05) greatly,while that of M2 macrophage which marked F4/80+CD11b+CD11c-was decreased(P<0.05)significantly.This trend was more remarkable in epididymal adipose tissue than in spleen(P<0.05).In epididymal adipose tissue,the percentage of CD4+T cells decreased but that of CD8+T cells increased in TLR4-/-mice.Moreover,the high-level expressions of IL-6,HMGB1,resistin were found in epididymal adipose tissue of TLR4-/-mice.However,the expressions of TNF-α and adiponectin decreased obviously.Conclusion: TLR4 knock-out could lead to a disorder in adipokine and immune cells in visceral adipose tissue.