摘要
目的研究氧化型低密度脂蛋白(ox-LDL)对巨噬细胞吞噬清除凋亡细胞的作用及其机制。方法 RAW264.7小鼠巨噬细胞随机分为对照组(无血清的DMEM培养液)、10μg/mL ox-LDL组(10μg/mL ox-LDL无血清DMEM培养液)和20μg/mLox-LDL组(20μg/mL ox-LDL无血清DMEM培养液),采用紫外光照射诱导RAW264.7小鼠巨噬细胞发生凋亡,流式细胞分析法检测RAW264.7小鼠巨噬细胞对凋亡细胞的吞噬指数,Western blotting和Real-Time PCR技术分别检测促吞噬受体MerTK蛋白和mRNA表达的变化。结果①孵育24 h后,10μg/mL ox-LDL组和20μg/mL ox-LDL组吞噬指数分别较对照组下降(4.7±2.8)%和(12.6±2.2)%,20μg/mL ox-LDL组显著小于对照组和10μg/mL ox-LDL组(P<0.05﹚。②孵育24 h后,10μg/mLox-LDL组和20μg/mL ox-LDL组MerTK蛋白表达灰度值分别较对照组下降(20.0±16.5)%和(47.0±15.4)%,20μg/mLox-LDL组显著小于对照组(P<0.05)。③孵育12 h后,10μg/mL ox-LDL组和20μg/mL ox-LDL组MerTK mRNA相对表达量分别较对照组减少(33.0±17.5)%和(60.0±10.0)%,10μg/mL ox-LDL组和20μg/mL ox-LDL组均显著小于对照组(P均<0.05)。结论 ox-LDL可抑制巨噬细胞对凋亡细胞的吞噬功能,其机制与抑制促吞噬受体MerTK的表达有关,这也可能是ox-LDL致动脉粥样硬化斑块不稳定和进展的机制之一。
Objective To investigate the effect and mechanism of macrophage phagocytosis of apoptotic cells（efferocytosis） treated by oxidized low-density lipoprotein（ox-LDL）.Methods RAW264.7 murine macrophages were randomly divided into control group（treated by serum-free DMEM culture medium）,10 μg/mL ox-LDL group（treated by 10 μg/mL ox-LDL serum-free DMEM culture medium） and 20 μg/mL ox-LDL group（treated by 20 μg/mL ox-LDL serum-free DMEM culture medium）.RAW264.7 murine macrophage apoptosis was induced by ultraviolet radiation,phagocytic index of RAW264.7 macrophages of apoptotic cells was determined by flow cytometry,and Western blotting and Real-Time PCR were employed to detect the expression of receptor MerTK protein and mRNA respectively.Results ①Twenty-four hours after treatment,the phagocytic indexes in 10 μg/mL ox-LDL group and 20 μg/mL ox-LDL group decreased by（4.7±2.8）% and（12.6±2.2）% respectively of that in control group,and the phagocytic index of 20 μg/mL ox-LDL group was significantly lower than that in control group and 10 μg/mL ox-LDL group（P〈0.05）.②Twenty-four hours after treatment,the relative expression of MerTK protein in 10 μg/mL ox-LDL group and 20 μg/mL ox-LDL group decreased by（20.0±16.5）% and（47.0±15.4）% respectively of that in control group,and the relative expression of MerTK protein in 20 μg/mL ox-LDL group was significantly lower than that in control group（P〈0.05）.③Twelve hours after treatment,the relative expression of MerTK mRNA in 10 μg/mL ox-LDL group and 20 μg/mL ox-LDL group decreased by（33.0±17.5）% and（60.0±10.0）% respectively of that in control group,and the relative expression of MerTK mRNA in 10 μg/mL ox-LDL group and 20 μg/mL ox-LDL group was significantly lower than that in control group（P〈0.05）.Conclusion ox-LDL can reduce the macrophage phagocytosis of apoptotic cells,and the mechanism may be related to inhibiting the expression of receptor MerTK,which may also be one of the causes for the inst
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2012年第6期706-710,715,共6页
Journal of Shanghai Jiao tong University:Medical Science
基金
国家自然科学基金(81000086)
上海市科委基金(2011-36059)~~
