双氢青蒿素对脂多糖诱导的心肌成纤维细胞分泌炎性因子及NF-κB通路激活的影响  

Effect of dihydroartemisinin on lipopolysaccharide induced cardiac fibroblasts inflammatory factors secretion and NF-κB pathway activation

作　　者：王燕[1] 单铁强[2] 栗志英[3] 李俊娇 马景红[3] 齐秀芳[4] WANG Yan;SHAN Tieqiang;LI Zhiying;LI Junjiao;MA Jinghong;QI Xiufang(Department of Comprehensive Surgery,Handan Second Hospital,Handan Hebei,056002,China;Department of Laboratory,Qinhuangdao Haigang Hospital,Qinhuangdao Hebei,066000,China;Internal Medicine-Cardiovascular Department,Affiliate Hospital of Hebei Engineering University,Handan Hebei,056002,China;Department of Hematology,Handan First Hospital,Handan Hebei,056002,China)

机构地区：[1]邯郸市第二医院综合外科,河北邯郸056002 [2]秦皇岛市海港医院检验科,河北秦皇岛066000 [3]河北工程大学附属医院心内科,河北邯郸056002 [4]邯郸市第一医院血液内科,河北邯郸056002

出　　处：《职业与健康》2023年第21期2907-2910,2916,共5页Occupation and Health

摘　　要：目的观察双氢青蒿素(dihydroartemisinin,DHA)对脂多糖刺激的心肌成纤维细胞(cardiac fibroblasts,CFs)增殖、分化、分泌胶原以及分泌炎性因子及核因子-κB(nuclear factor-κB,NF-κB)通路激活的影响,探究DHA抑制心脏纤维化的分子机制。方法采用胰酶消化SD乳鼠心肌组织,获取其中的CFs。实验将CFs分为正常组(培养液)、DHA组(100 ng/mL DHA的培养液)、脂多糖组(50μg/mL脂多糖的培养液)、治疗组(50μg/mL脂多糖和100 ng/mL DHA的培养液)。每组细胞同时孵育48 h后,检测各组CFs的增殖变化;ELISA定量培养液中Ⅰ型胶原(collagen type I,ColⅠ)、Ⅲ型胶原(collagen typeⅢ,ColⅢ)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白介素-6(interleukin-6,IL-6)及白介素-1β(interleukin-1β,IL-1β)的水平;免疫印迹法评估胞质内平滑肌-a-肌动蛋白(a-smooth muscle actin,a-SMA)、核因子κBp65(nuclear factorκBp65,NF-κBp65)及磷酸化核因子κBp65(phosphorylated nuclear factorκBp65,p-NF-κBp65)的水平。结果与正常组对比,DHA组的细胞增殖、培养液中ColⅠ、ColⅢ、TNF-α、IL-6及IL-1β的含量,胞质内a-SMA、NF-κBp65及p-NF-κBp65蛋白含量及NF-κBp65/p-NF-κBp65比值比较差异均无统计学意义(均P>0.05);脂多糖组细胞数目(0.1229±0.0014)增加,培养液中ColⅠ[(0.7532±0.0877)ng/L]、ColⅢ[(0.7233±0.0968)ng/L]、TNF-α[(46.78±3.60)pg/L]、IL-6[(30.08±3.93)pg/L]及IL-1β[(12.10±1.55)pg/L]含量均上升,胞质内a-SMA(0.438±0.026)、NF-κBp65(1.221±0.049)、p-NF-κBp65(1.209±0.048)的含量及NF-κBp65/p-NF-κBp65比值(0.967±0.046)均增加(均P<0.05);与脂多糖组对比,治疗组的细胞数目(0.1156±0.0012)减少,培养液中ColⅠ[(0.5424±0.0636)ng/L]、ColⅢ[(0.5027±0.0579)ng/L]、TNF-α[(25.32±3.05)pg/L]、IL-6[(19.12±3.00)pg/L]及IL-1β[(7.34±0.96)pg/L]含量均下降,胞质内a-SMA(0.208±0.020)、NF-κBp65(0.528±0.035)、p-NF-κBp65(0.206±0.026)的含量及NF-κBp65/p-NF-κBp65比值(0.412±0.025)均减少(Objective To observe the effects of dihydroartemisinin(DHA)on the proliferation,differentiation,collagen secretion,inflammatory factors and NF-κB pathway activation of lipopolysaccharide induced cardiac fibroblasts(CFs),explore the molecular mechanism of DHA inhibiting cardiac fibrosis.Methods The myocardial tissue of SD suckling rats was digested with trypsin to obtain CFs.CFs were arranged into normal group(culture medium),DHA group(culture medium of 100 ng/mL DHA),lipopolysaccharide group(50μg/mL lipopolysaccharide culture medium),the treatment group(50μg/mL lipopolysaccharide and 100 ng/mL DHA).After 48 h incubation of cells in each group,the proliferation of CFS in each group was detected.The ELISA was used to quantify the level of collagen typeⅠ(ColⅠ),collagen typeⅢ(ColⅢ),tumor necrosis factorα(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β)in the culture medium.The immunoblotting was used to evaluate intracellular smooth muscle-a-actin(a-SMA),nuclear factorκB p65(NF-κBp65),and phosphorylated nuclear factorκB p65(p-NF-κBp65).Results The cell proliferation,the content of colⅠ,colⅢ,TNF-α,IL-6 and IL-1βin suspension,protein content of a-SMA,NF-κBp65 and p-NF-κBp65 and NF-κBp65/p-NF-κBp65 ratio in intracytoplasmbetween DHA group and normal group had not obviously difference(all P>0.05).Compared with the normal group,the number of cells in lipopolysaccharide group(0.1229±0.0014)increased,and the contents of colⅠ[(0.7532±0.0877)ng/L],colⅢ[(0.7233±0.0968)ng/L],TNF-α[(46.78±3.60)pg/L],IL-6[(30.08±3.93)pg/L]and IL-1β[(12.10±1.55)pg/L]in suspension increased,and the intracytoplasm contents of a-SMA(0.438±0.026),NF-κBp65(1.221±0.049),p-NF-κB p65(1.209±0.048)and NF-κBp65/p-NF-κBp65 ratio(0.967±0.046)increased(all P<0.05).Compared with the lipopolysaccharide group,the number of cells(0.1156±0.0012)decreased in treatment group,and the levels of colⅠ[(0.5424±0.0636)ng/L],colⅢ[(0.5027±0.0579)ng/L],TNF-α[(25.32±3.05)pg/L],IL-6[(19.12±3.00)pg/L]and IL-1β[(7.34±

关 键 词：双氢青蒿素 脂多糖 心肌成纤维细胞 炎性因子 NF-ΚB通路 

分 类 号：R114[医药卫生—卫生毒理学]