知识与财富的链接
目的：探讨白藜芦醇对脂多糖（LPS）诱导的小鼠巨噬细胞系RAW264．7细胞极化表型改变的影响。方法体外培养的RAW264．7小鼠巨噬细胞在六孔板孵育12 h ,分为磷酸盐缓冲液（PBS）对照组、LPS（100 ng／mL）组、LPS（100 ng／mL）＋白藜芦醇（30μmol／L）组,LPS＋白藜芦醇组在加入LPS前预先加入白藜芦醇孵育12 h;上述细胞在LPS刺激12 h后分别收集细胞和培养上清液;实时定量PCR（RT‐qPCR）检测经典的炎症活化型巨噬细胞（M1）型相关基因iNOS、TNF‐αmRNA表达,以及替代活化巨噬细胞（M2）型相关基因IL‐10、PPARγ和Arg‐1 mRNA表达,Western blot检测细胞iNOS、Arg‐1蛋白表达,ELISA检测培养上清液中炎性因子IL‐12 p40、IL‐10和 TNF‐α的水平。结果 RT‐qPCR检测结果显示,与LPS组相比,M1型相关基因iNOS、TNF‐αmRNA较LPS＋白藜芦醇组显著上升（P＜0．05）,而M2型相关基因IL‐10、PPARγ、Arg‐1 mRNA表达显著下调（P＜0．05）。Western blot检测发现,LPS组iNOS蛋白水平显著高于LPS＋白藜芦醇组（P＜0．05）,而Arg‐1蛋白水平显著低于LPS＋白藜芦醇组（P＜0．05）。ELISA检测发现,LPS组培养上清液中炎性因子IL‐12 p40、TNF‐α水平显著高于LPS＋白藜芦醇组（P＜0．05）,而IL‐10和水平显著低于LPS＋白藜芦醇组（P＜0．05）。结论白藜芦醇可能促进了LPS刺激的RAW264．7巨噬细胞向M2型极化改变。
Objective To investigate the effect of resveratrol on murine macrophage cell line (RAW264 .7 cells) polarizing phenotype induced by lipopolysaccharide .Methods RAW264 .7 mouse macrophages seeded in a 6 well plate ,then randomly divided into phosphate buffer saline(PBS) control group ,LPS (100 ng/mL) group ,and LPS (100 ng/mL)+ resveratrol (30 μmol/L) group .In the LPS+ resveratrol group ,LPS was added after incubation with resveratrol for 12 h .Cells were harvested and superna‐tant were collected after incubation with LPS for 12 h .Both the mRNA expression levels of M1 associated markers iNOS and TNF‐αand M2 associated markers IL‐10 ,PPARγ and Arg‐1 were measured by real time quantitative PCR .Expression of iNOS ,Arg‐1 protein were detected by Western blot ,inflammatory factor IL‐12 p40 ,IL‐10 and TNF‐αprotein in the supernatant of were assayed by ELISA .Results PCR detection showed that the mRNA expression levels of M1 associated markers iNOS and TNF‐αin the LPS group were significantly higher than that of LPS+ resveratrol group(P<0 .05) ,but the mRNA expression levels of M2 associated markers IL‐10 ,PPARγand Arg‐1 were significantly lower than that of LPS+ resveratrol group(P<0 .05) .Compared with LPS+resveratrol group ,western blot assay showed that iNOS protein level in LPS group was significantly higher than it (P<0 .05) ,but Arg‐1 protein level was significantly lower than it(P<0 .05) .The levels of IL‐12 p40 and TNF‐αin LPS group were significantly higher than that in LPS+ resveratrol group(P<0 .05) ,but the levels of IL‐10 was significantly lower than it(P<0 .05) .Conclusion Resveratrol may promote LPS stimulated RAW264 .7 macrophage polarization to M2 phenotype .
