岩黄连总碱对巨噬细胞M1型分化的抑制作用  

Inhibitory effects of Corydalis saxicola Bunting total alkaloid on M1 macrophages polarization

作　　者：冯葱 雍翔智[1,2,3,4] 江巧芝 吴恬恬 蒋兰岚 苏志恒 陶人川[1,2,3,4] Cong Feng;Xiangzhi Yong;Qiaozhi Jiang;TiantianWu;Lanlan Jiang;Zhiheng Su;Renchuan Tao(College&Hospital of Stomatology,Guangxi Medical University;Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction;Guangxi Clinical Research Center for Craniofacial Deformity;Guangxi Key Laboratory of Oral and Maxillofacial Surgery-Disease Treatment;Pharmaceutical College,Guangxi Medical University)

机构地区：[1]广西医科大学附属口腔医院 [2]广西口腔颌面修复与重建研究重点实验室 [3]广西颅颌面畸形临床医学研究中心 [4]广西口腔颌面外科疾病治疗重点实验室 [5]广西医科大学药学院,南宁530021

出　　处：《广西医科大学学报》2020年第12期2103-2110,共8页Journal of Guangxi Medical University

基　　金：This research was supported by grants from the National Natural Science Foundation of China(81771073);Innovation Project of Guangxi Graduate Education(YCSW2020127);Youth Science Foundation of Guangxi Medical University(GXMUYSF201813).

摘　　要：目的:研究岩黄连总碱(CSBTA)对巨噬细胞M1型分化的影响。方法:THP-1细胞经佛波脂(PMA)刺激48 h诱导分化为巨噬细胞(Mφ)。倒置显微镜观察细胞形态变化,流式细胞术检测巨噬细胞表面分子CD11b、CD14;加入脂多糖(LPS,1μg/mL)、γ干扰素(IFN-γ,20 ng/mL)刺激48 h诱导Mφ巨噬细胞分化为M1型巨噬细胞。不同浓度(0.0025 g/L-0.02 g/L)CSBTA干预M1型巨噬细胞24 h。采用实时荧光定量PCR(RT-qPCR)法检测肿瘤坏死因子(TNF)-α和白细胞介素(IL)-6 mRNA相对表达量,Western blotting法检测M1型巨噬细胞CD86蛋白表达量,酶联免疫吸附试验(ELISA)法检测细胞培养上清液中IL-1β的含量。结果:0.02 g/L的CSBTA对M1型巨噬细胞活力产生明显的毒性作用(P<0.05)。在0.0025 g/L、0.005 g/L CSBTA的干预下,M1型巨噬细胞IL-6和TNF-αmRNA相对表达量、CD86蛋白表达量及IL-1β含量均显著降低(均P<0.05)。结论:CSBTA可以有效抑制THP-1源性巨噬细胞M1型的分化,并改善炎症环境。Objective: To investigate the effects of Corydalissaxicola Bunting total alkaloid (CSBTA) on classically activatedtype 1 (M1) polarization of macrophages (Mφ). Methods:THP-1 cells were induced to differentiate into Mφ by phorbolester (PMA) for 48 h. Morphological change of the cells wasobserved using inverted microscope. Flow cytometry was appliedto detect the myloid cell markers CD11b and CD14, aswell as the major marker of Mφ. CSBTA was given to M1-Mφwhich was induced from M0-Mφ by interferon (IFN-γ, 20 ng/mL) and lipopolysaccharide (LPS, 1 μg/mL), at various concentrationsranging from 0.002 5 g/L to 0.02 g/L for 24 h. ThemRNA levels of tumor necrosis factor (TNF)-α and interleukin(IL)- 6 were detected by quantitative real- time PCR (RT- qPCR).The expression level of CD86 was determined by westernblotting. The amount of IL- 1β in the cell culture supernatantwas measured by enzyme linked immunosorbent assay (ELISA).Results: 0.02 g/L of CSBTA had a significant toxic effecton the vitality of M1-Mφ (P<0.05). 0.002 5 g/L and 0.005 g/Lof CSBTA significantly reduced the mRNA levels of IL-6 andTNF-α (P<0.05), the protein expression of CD86 in M1-Mφand the production of IL-1β in the cell culture supernatant (P<0.05). Conclusion: CSBTA can effectively suppress M1 polarizationof THP-1-derived Mφ that may improve the inflammatoryenvironment.

关 键 词：岩黄连总碱 巨噬细胞 分化 

分 类 号：R114[医药卫生—卫生毒理学]