脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

IL-10对脂多糖诱导Ana-1细胞的MyD88/核因子κB活性影响的研究

目的探讨IL-10对小鼠巨噬细胞髓样分化因子88(MyD88)/核因子κB(NF-κB)炎症信号活化的影响。方法将小鼠巨噬细胞Ana-1分为脂多糖(LPS)组和LPS+IL-10组,分别于0.5、1及2h收集巨噬细胞和细胞培养上清液,Western blot检测细胞MyD88与胞浆、胞核NF-κBp65亚基表达,ELISA法检测培养上清中肿瘤坏死因子α(TNF-α)含量。结果在0～2h,LPS组细胞MyD88表达显著持续上升,LPS+IL-10组于LPS刺激后上升,0.5h达峰值,2h恢复至正常水平,1h和2h相对含量均低于LPS组(11.6±1.3比17.5±0.7,8.8±0.3比21.4±1.8,P0.05);总NF-κB表达量在两组间无明显差异。NF-κB核浆比变化趋势与MyD88类似,LPS+IL-10组1h及2h相对含量亦均低于LPS组(1.1±0.1比2.4±0.4,0.6±0.7比3.1±0.6,P0.05);相应的,LPS+IL-10组1h和2hTNF-α含量亦低于LPS组[(222.5±33.5)pg/mL比(365.2±22.7)pg/mL,(212.7±15.9)pg/mL比(566.2±31.5)pg/mL,P0.05]。结论 IL-10通过抑制减少MyD88/NF-κB信号通路活化,降低TNF-α表达,从而下调炎症反应强度。     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

吉非替尼对博莱霉素致肺纤维化小鼠α平滑肌肌动蛋白的抑制作用

目的 观察吉非替尼对博莱霉素诱导肺纤维化小鼠a平滑肌肌动蛋白(a-SMA)表达的影响,探讨EGFR在肺纤维化上皮间质转分化中的作用.方法 将30只SPF级雌性BALB/c小鼠分为三组:对照组(气管滴人生理盐水)、纤维化组(气管滴人博莱霉素3 mg/kg )、纤维化吉非替尼干预组(气管滴人博莱霉素+吉非替尼灌胃20 mg/Kg ).实验第14天杀鼠取肺,肺组织石蜡切片行HE染色与Masson染色;RT-PCR法检测a-SMA的mRNA表达水平;免疫组化检测总EGFR,磷酸化EGFR及a-SMA表达.结果 纤维化吉非替尼干预组肺病理损伤较纤维化组减轻,气道上皮下胶原沉积减少,气道上皮及肺间质细胞磷酸化EGFR表达评分下降(P<0.05);同时,肺a-SMA的mRNA表达减少,免疫组化评分也明显降低(P<0.05).结论 吉非替尼抑制博莱霉素诱导小鼠肺纤维化可能与其抑制EGFR活性,下调a-SMA表达有关.     

NFκB1基因启动序列-94ins／delATTG多态性与社区获得性肺炎易感性及严重程度相关性研究

目的探讨NFκBl基因启动序列-94ins／del ATTG基因多态性与中国广东省汉族人群社区获得性肺炎（CAP）易感性及严重程度的相关性。方法采用聚合酶链反应-高分辨率熔解曲线法（PCR—HRM）检测66例CAP患者和66例健康体检者的NFκB1基因启动序列-94ins／de1 ATTG基因型的分布,分析该基因多态性与CAP易感性及严重性的相关性。结果NFxB1基因启动序列-94ins／del ATTG基因型和等位基因分布频率在CAP组和健康对照组比较差异无统计学意义（P〉0．05）;在重症肺炎组与非重症肺炎组比较差异无统计学意义（P〉O．05）;NFκB1-94ins／delATTG的不同基因型与CAP患者的性别、年龄、住院时间、白细胞数、中性粒细胞数之间的相关性均无统计学意义（P〉0．05）。结论NFκB1基因启动序列-94ins／delATTG多态性与cAP易感性及严重程度无相关性。     

小剂量阿奇霉素对支气管哮喘患者气道炎症的影响

目的 探讨支气管哮喘患者常规规律吸入布地奈德福莫特罗粉吸入剂同时叠加口服小剂量阿奇霉素对气道炎症的疗效.方法 选择2018年7月至2020年5月就诊于解放军南部战区空军医院的200例轻、中度支气管哮喘患者,采用随机数字表法分为对照组与治疗组,每组100例,所有研究对象均规律吸入布地奈德福莫特罗粉吸入剂,治疗组叠加口服小...     

NF-κB p65基因在TNF-α诱导的肺泡上皮细胞氧化损伤中的作用

目的建立急性肺损伤肺泡上皮细胞炎症模型,探讨NF-κB p65基因在炎症诱导的氧化应激损伤中的作用。方法以肿瘤坏死因子α（TNF-α,10 ng/mL）刺激A549细胞,运用RNA干扰技术沉默核因子κB（NF-κB）p65基因,采用RT-PCR及Western blot法检测沉默效率,ELISA法检测细胞培养上清中白细胞介素1β（IL-1β）、IL-4、IL-6等炎症因子浓度,比色法检测细胞内丙二醛（MDA）及超氧化物歧化酶（SOD）浓度,MTT法检测细胞存活率。结果 TNF-α刺激A549细胞可在基因及蛋白水平上调NF-κB p65的表达,并增加NF-κB蛋白的核转位;同时细胞培养上清中IL-1β、IL-4、IL-6的浓度升高,细胞内MDA增多,SOD减少,细胞存活率降低。预转染NF-κB p65 siRNA可在基因水平及蛋白水平有效沉默NF-κB p65表达,降低TNF-α诱导的上述各炎症因子及MDA浓度的增高,减少SOD的耗损,A549细胞存活率升高（P〈0.05）。结论 NF-κB能介导TNF-α触发的过度炎症反应和氧化应激,导致细胞存活率明显降低;沉默NF-κB p65基因可有效下调炎症反应水平及其诱发的氧化应激,减轻肺结构细胞的损伤。     

美国感染疾病学会2009季节性流行性感冒临床指南简介

在世界范围内每年均有一定数量的人群死于季节性流行性感冒(以下简称流感),由于预计2009年有全球性流感暴发的可能性,美国感染病学会(IDSA)于2009年4月公布"成人和儿童季节性流行性感冒的诊断、治疗、预防和公共机构暴发的处理指南"~([1])(以下简称"指南"),详细地论述了季节性流感的诊断、治疗与预防,为流感的处理提供了临床操作规范,现介绍如下.     

肺纤维化治疗新思路——EGFR-TKI

肺纤维化属于纤维增殖性疾病的范畴,目前尚无有效的治疗手段,进行性呼吸衰竭、并发肺肿瘤或心血管疾病而死亡是肺纤维化疾病的转归。目前研究表明表皮生长因子(Epidermal growth factor,EGF)家族及其受体(EGF receptor,EGFR)在肺纤维化中高表达,EGFR酪氨酸激酶抑制剂(EGFR tyrosine kinase inhibitor,EGFR-TKI)能抑制EGFR的胞内区酪氨酸激酶活性,阻断纤维化信号,减少成纤维细胞活化。动物实验结果已证实EGFR-TKI能缓解肺纤维化改变。基于EGFR-TKI有缓解肺纤维化的潜力,同时耐受性好,不良反应小,有望成为新一代抗纤维化药物。