穿心莲内酯对临床铜绿假单胞菌感染人支气管上皮细胞的干预研究

Effects of andrographolide on infection of human bronchial epithelial cells caused by clinical strains of pseudomonas aeruginosa

目的:用铜绿假单胞菌(pseudomonas aeruginosa,PA)感染人支气管上皮细胞BEAS-2B,观察穿心莲内酯对细菌感染性肺炎及其诱发的宿主细胞炎性反应的影响。方法:PA标准株PAO1和临床分离株Cq1(exoU-)及Cq40(exoU+)感染上皮细胞BEAS-2B;体外肉汤培养观察穿心莲内酯(浓度300μmol/L)对细菌生长的影响。侵袭实验、乳酸脱氢酶(lactic dehydrogenase,LDH)释放实验、菌落计数法检测PA对BEAS-2B细胞的侵袭力;ELISA法检测BEAS-2B细胞炎症因子白细胞介素(interleukin,IL)-6、IL-8的表达水平。穿心莲内酯(3.0μmol/L和4.5μmol/L)用于对PA感染人BEAS-2B的干预实验。结果:同PAO1相似,PACq1及Cq40在LB培养基中的生长均不受高浓度(300μmol/L)穿心莲内酯影响。但穿心莲内酯(3.0μmol/L和4.5μmol/L)降低侵入胞内的PA数,减少细菌感染所致的上皮细胞LDH释放,也降低了细菌刺激上皮细胞IL-6及IL-8的表达水平。结论:穿心莲内酯可降低PA临床分离株对人支气管上皮细胞的侵袭力,并可降低细胞损伤及抑制细胞炎症反应。

Objective：To infect human bronchial epithelial cell （BEAS-2B） model by pseudomonas aeruginosa（PA） and to observe andrographolide＇s effects on the bacterial infection ability and the host cell inflammatory response. Methods ：PA01 and clinical strains of PA,Cql （exoU-） and Cq40（exoU＋） were respectively used to infect BEAS-2B. In vitro lysogeny broth culture was conducted and the effect of andrographolide（300 Ixmol/L） on the bacterial growth was observed. PA invasive capability on BEAS-2B was tested by colony counting method. With double antibody sandwich enzyme-linked immunosorbent assay（ELISA）,lactate dehydrogenase（LDH） releasing and inflammatory cytokines, interleukin-6（IL-6） and IL-8 expression were measured. Pretreated by different concentrations of andrographolide （3.0 umol/L and 4.5 umol/L）, effects of andrographolide on BEAS-2B infection caused by different strains of PA were detected. Results：Similar to PAO1 ,high concentration （300 umol/L） andrographolide exerted no impact on either e xoU＋ or exoU-PA growth. 3.0 u^mol/L and 4.5 b^mol/L andrographolide significantly reduced the intraeellular colony number of PA, LDH re leasing and generation of IL-6 and IL-8. Conclusions： Andrographolide can attenuate invasive effects of clinical strains of PA on hu- man bronchial epithelial cell and reduce the related cell damage and inhibit inflammatory response.