(1.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; 2.临床医学系; 3.基础医学国家级实验教学示范中心;徐州医科大学,江苏徐州 221004)
R-33
(1. Jiangsu Key Laboratory of Immunity and Metabolism, Department of Pathogenic Biology and Immunology;2.Department of Clinical Medicine; 3. National Experimental Teaching Demonstration Center for Basic Medicine;Xuzhou Medical University, Xuzhou 221004, China)
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 tensionhomology 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(20 μg/ mL)?LPS(100 ng/ mL)?PPC(20 μg/ mL) + LPS(100 ng/mL)?PPC(20 μg/ mL)+ SF1670(PTEN 抑制剂,150 ng/ mL)?PPC(20 μg/ 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;RTPCR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 onchromosome ten (PTEN) is involved in the process. The study also preliminarily determined the anti-inflammatory effect ofPPC and its underlying mechanisms. Methods Raw 264. 7 cells were inoculated on cell culture plates and divided into sixgroups, which were labeled and induced as follows: PPC (20 μg/ mL), LPS (100 ng/ mL), LPS (100 ng/ mL) + PPC(20 μg/ mL), PPC (20 μg/ mL) + SF1670 (150 ng/ mL), PPC (20 μg/ 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 detectthe mRNA expression of IL-6. Western blot was performed to confirm the PTEN protein expression. Results TNF-α andIL-10 levels in the supernatants did not significantly differ between the PBS and PPC groups. Compared with the LPSgroup, the TNF-α and 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 significantlyhigher than that of the LPS group. Compared with the PPC group, the TNF-α levels and IL-6 mRNA expression in the PPC+SF1670 group were significantly increased ( P <0. 05). The PPC+LPS+SF1670 group showed elevated TNF-α and IL-6production ( 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.
姜文清,戚艳,沙若荷,章欣,陈静越,潘伟,孙芬芬. 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[J].中国比较医学杂志,2019,29(8):44~49.
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