PTEN在多烯磷脂酰胆碱下调LPS诱导巨噬细胞炎症反应中的抑制作用  

Inhibitory effect of PTEN on polyene phosphatidylcholine in downregulating LPS-induced macrophage inflammatory response

作　　者：姜文清 戚艳 沙若荷 章欣 陈静越 潘伟 孙芬芬 JIANG Wenqing;QI Yan;SHA Ruohe;ZHANG Xin;CHEN Jingyue;PAN Wei;SUN Fenfen(Xuzhou Medical University,Xuzhou 221004,China;Department of Clinical Medicine,Xuzhou Medical University,Xuzhou 221004,China;National Experimental Teaching Demonstration Center for Basic Medicine)

机构地区：[1]徐州医科大学江苏省免疫与代谢重点实验室病原生物学与免疫学教研室,江苏徐州221004 [2]徐州医科大学临床医学系,江苏徐州221004 [3]徐州医科大学基础医学国家级实验教学示范中心,江苏徐州221004

出　　处：《中国比较医学杂志》2019年第8期44-49,共6页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金(81871670);国家级大学生创业创新训练计划项目(201710313023Z,201710313023);江苏省博士后科研资助计划(2019K063)

摘　　要：目的研究临床护肝药—多烯磷脂酰胆碱(polyene phosphatidyl choline,PPC)对LPS诱导巨噬细胞(macrophages,M)炎症的调控作用以及第10号染色体上缺失的磷酸酶-张力蛋白同系物(phosphate and tension homology deleted on chromosome ten,PTEN)在上述过程中的作用,初步明确PPC的抗炎效果及机制。方法将Raw264.7细胞铺于细胞培养板,分别加入PPC(20μg/mL)、LPS(100 ng/mL)、PPC(20μg/mL)+LPS(100 ng/mL)、PPC(20μg/mL)+SF1670(PTEN抑制剂,150 ng/mL)、PPC(20μg/mL)+LPS(100 ng/mL)+SF1670(150 ng/mL)以及等体积PBS,培养24 h后,收集细胞沉淀及培养上清,ELISA法检测上清中IL-6、IL-10、TNF-α含量;RT-PCR检测细胞中IL-6的mRNA相对表达量;Western blot检测细胞中PTEN的蛋白表达情况。结果相比PBS组,PPC组TNF-α、IL-6、IL-10分泌量无明显差异;相比LPS组,PPC+LPS组培养上清中TNF-α、IL-6水平明显降低(P<0.001),而IL-10水平明显升高(P<0.001)。相比LPS组,PPC+LPS组PTEN蛋白表达量明显上高。相比PPC组,PPC+SF1670组TNF-α、IL-6分泌或表达量明显升高(P<0.05);相比PPC+LPS组,PPC+LPS+SF1670组TNF-α、IL-6分泌或表达水平明显升高(P<0.05),而IL-10分泌量明显降低(P<0.05)。结论PPC可通过上调PTEN表达以抑制LPS诱导M炎症反应。Objective To investigate the regulatory role of the clinical hepatinica,polyene phosphatidylcholine(PPC),on LPS-induced macrophage inflammatory response and whether phosphate and tension homology deleted on chromosome ten(PTEN)is involved in the process.The study also preliminarily determined the anti-inflammatory effect of PPC and its underlying mechanisms.Methods Raw 264.7 cells were inoculated on cell culture plates and divided into six groups,which were labeled and induced as follows:PPC(20μg/mL),LPS(100 ng/mL),LPS(100 ng/mL)+PPC(20μg/mL),PPC(20μg/mL)+SF1670(150 ng/mL),PPC(20μg/mL)+SF1670(150 ng/mL)+LPS(100 ng/mL)or an equal volume of phosphate-buffered saline(PBS)groups.The cells and supernatants were collected after 24 h.Enzyme-linked immunosorbent assay was used to detect the levels of IL-6,IL-10 and TNF-α.RT-PCR was applied to detect the mRNA expression of IL-6.Western blot was performed to confirm the PTEN protein expression.Results TNF-αand IL-10 levels in the supernatants did not significantly differ between the PBS and PPC groups.Compared with the LPS group,the TNF-αand IL-6 levels in culture supernatants of the PPC+LPS group decreased significantly(P<0.001),while the IL-10 level increased significantly(P<0.001).PTEN protein expression in the PPC+LPS group was significantly higher than that of the LPS group.Compared with the PPC group,the TNF-αlevels and IL-6 mRNA expression in the PPC+SF1670 group were significantly increased(P<0.05).The PPC+LPS+SF1670 group showed elevated TNF-αand IL-6 production(P<0.001)but lower IL-10 production(P<0.001)than that of the PPC+LPS group.Conclusions PPC inhibits LPS-induced macrophage inflammatory response by upregulating PTEN expression.

关 键 词：多烯磷脂酰胆碱 巨噬细胞 PTEN 炎症因子 

分 类 号：R-33[医药卫生]