摘要
目的观察中链脂肪酸辛酸(C8:0)对RAW264.7巨噬细胞ABCA1表达及JAK2/STAT3通路相关因子的影响。方法建立不同脂肪酸分组:空白组,C8:0组,EPA组和cAMP组。干预RAW264.7巨噬细胞24 h后,采用逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)检测其ABCA1、JAK2/STAT3通路相关因子mRNA的表达,蛋白质印迹法(western blot,WB)检测ABCA1、JAK2/STAT3通路相关因子蛋白的表达,酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)检测TNF-α、IL-6、MCP-1水平。结果C8:0200μmol/L较100μmol/L能更显著增高RAW264.7巨噬细胞ABCA1 mRNA和蛋白的表达(P<0.05),确定后续脂肪酸干预浓度为200μmol/L;在200μmol/L浓度下,与空白组相比,C8:0组ABCA1、STAT3的mRNA表达量显著增高,而P65的mRNA表达量显著降低(P均<0.05);且C8:0组ABCA1、p-STAT3及p-JAK2蛋白表达量显著增高,而P65蛋白表达量显著降低(P均<0.05);C8:0组炎性因子TNF-α显著低于空白组,IL-6水平显著低于cAMP组,MCP-1水平显著低于EPA组(P均<0.05)。结论中链脂肪酸C8:0可能通过上调ABCA1抑制RAW264.7巨噬细胞JAK2/STAT3通路的炎症表达。
Objective To observe the effect of the medium-chain fatty acid octanoic acid(C8:0)on the expression of ABCA1 and JAK2/STAT3 inflammatory pathway in RAW264.7 macrophages.Methods Fatty acids with different concentrations were used to establish different experiment groups:negative control group,C8:0 group,C16:0 group and cAMP group.After 24 hours of intervention in RAW264.7 macrophages,relative expression of ABCA1 mRNA and JAK2/STAT3 pathway-related mRNA was detected by Reverse Transcription-Polymerase Chain Reaction(RT-PCR),relative level of ABCA1/JAK2/STAT3 pathway-related protein was detected by western blot(WB),and TNF-ɑ,IL-6 and MCP-1 were detected by enzyme linked immunosorbent assay(ELISA).Results The relative expressions of ABCA1 mRNA and protein in RAW264.7 macrophages were significantly higher at C8:0200μmol/L than those at 100μmol/L(P<0.05).At the concentration of 200μmol/L,compared with the negative control(NC)group,the relative mRNA expression levels of ABCA1 and STAT3 in C8:0 group significantly increased,while the relative mRNA expression levels of P65 decreased significantly(all P<0.05).Meanwhile,the relative expression levels of ABCA1,p-STAT3 and p-JAK2 in the C8:0 group increased significantly,while the relative expression levels of P65 decreased significantly(all P<0.05).The inflammatory factor level of TNF-ɑin C8:0 group was significantly lower than that in negative control group,the level of IL-6 was significantly lower than that in cAMP group,and the level of MCP-1 was significantly lower than that in EPA group(all P<0.05).Conclusion C8:0 can inhibit inflammatory expression by up-regulating ABCA1 and inhibit JAK2/STAT3 pathway in RAW264.7 macrophages.
作者
张碰
张新胜
刘玉猛
陶扬
于志盟
刘鹿
刘英华
ZHANG Peng;ZHANG Xinsheng;LIU Yumeng;TAO Yang;YU Zhimeng;LIU Lu;LIU Yinghua(Department of Nutrition,the First Medical Center,Chinese PLA General Hospital,Beijing 100853,China)
出处
《解放军医学院学报》
CAS
2020年第5期513-517,共5页
Academic Journal of Chinese PLA Medical School
基金
国家自然科学基金(81703204)
