摘要
[目的]探讨在游离SiO_2诱导大鼠肺泡巨噬细胞泡沫化过程中抑制过氧化物酶增殖体激活性受体(PPARγ)对CD36表达以及脂质蓄积的调节作用。[方法]常规体外培养大鼠肺泡巨噬细胞株NR8383细胞,给予PPARγ特异性抑制剂GW9662阻断PPARγ信号通路,观察细胞的泡沫化情况;细胞分为抑制剂组(GW9662)、溶剂对照组(DMSO)、实验组(GW9662+SiO_2+ox-LDL)、模型组(DMSO+SiO_2+ox-LDL),培养36 h。油红O染色镜下观察脂质蓄积情况;酶法测定细胞中总胆固醇、游离胆固醇表达情况,计算胆固醇酯占总胆固醇的比例,鉴定泡沫细胞的形成;应用Western blot和实时荧光定量聚合酶链式反应技术分别检测PPARγ和CD36的蛋白和基因表达情况。[结果]实验结果显示,与模型组[(31.41±1.36)mg/g]比较,实验组[(8.78±0.49)mg/g]、抑制剂组[(6.23±0.27)mg/g]和溶剂对照组[(6.99±0.30)mg/g]细胞内总胆固醇含量明显降低(P<0.05);且实验组(28.43%)、抑制剂组(27.02%)和溶剂对照组(24.92%)的胆固醇酯占总胆固醇比例均小于50%,3组之间差异没有统计学意义(P>0.05);与模型组相比,实验组泡沫细胞数目明显减少。实验组CD36蛋白(0.55±0.13)和m RNA(1.30±0.39)表达也较模型组[分别为(1.08±0.31)和(3.38±0.70)]明显降低(P<0.05)。[结论]游离SiO_2所诱导的CD36表达可能是由PPARγ介导所致。
[ Objective ] To investigate the regulatory effect of inhibiting peroxisome proliferator-activated receptor gamma （PPARY） on CD36 expression and lipid accumulation during the foaming of alveolar macrophages induced by free silica （SiO2）. [ Methods ] Rat alveolar macrophage strain NR8383 cells were cultured in vitro. The PPARγ pathway was blocked by inhibitor GW9662. The foaming of the ceils was observed. The cells were divided into inhibitor （GW9662）, solvent control （DMSO）, e.xperiment （GW9662＋SiO2＋ox-LDL）, and model （DMSO＋SiO2＋ox-LDL） groups, and cultured for 36 h. Oil red O staining was used to observe the accumulation of lipid. The expressions of intracellular total cholesterol and free cholesterol were detected by enzyme assay to calculate the ratio of cholesterol ester to total cholesterol and identify the formation of foam cells. Western blot and real-time fluorescence PCR were used to detect the protein and gene expressions of PPARγ and CD36.[ Results ] The total cholesterol levels in the experiment group [（8.78 ± 0.49） rag/g], inhibitor group [（6.23 ± 0.27） mg/g], and solvent control group [（6.99± 0.30）mg/g] were significantly lower than that in the model group [（31.41± 1.36） mg/g] （P 〈 0.05）. The cholesterol ester/total cholesterol ratio was less than 50% in the experiment group （28.43%）, inhibitor group （27.02%）, and solvent control group （24.92%）, and there was no difference among the three groups （P〉0.05）. The number of foam cells in the experiment group was significantly reduced compared with the model group. Both the CD36 protein （0.55 ± 0.13） and mRNA （1.30 ± 0.39） expression levels of the inhibitor group were significantly lower lower than those of the model group [（1 08 ± 0 31）, （3.38 ± 0.70）] （P〈0.05）. [ Conclusion ] CD36 expression induced by free SiO2 may be mediated by PPARγ.
出处
《环境与职业医学》
CAS
CSCD
北大核心
2018年第1期55-59,共5页
Journal of Environmental and Occupational Medicine
基金
国家自然科学基金资助项目(编号:81602814)
河北省卫计委医学科学研究重点课题计划(编号:20160720)
唐山市科技计划项目(编号:15130212C)
研究生创新项目(编号:2017S10)
