摘要
目的探讨p38MAPK在二烯丙基三硫(DATS)抑制脂多糖(LPS)诱导小鼠肺泡巨噬细胞促炎细胞因子表达中的作用。方法体外培养MH-S细胞,用DATS和(或)LPS进行干预,Western blot检测细胞p38及磷酸化p38(p-p38)的表达;用LPS和(或)SB203580孵育细胞,反转录PCR检测细胞中TNF-α、IL-1βmRNA表达,Western blot检测细胞磷酸化(p-IκB)及非磷酸化IκB的表达。结果 LPS刺激MH-S细胞可导致p-p38表达增加,呈时间依赖性;用DATS(0.1、0.5、2.5、5.0 mg.L-1)预处理细胞30 min后再给予LPS刺激,p-p38表达呈剂量依赖性下降;单独DATS对p-p38表达无明显影响。p38特异性抑制剂SB203580可剂量依赖性地抑制LPS诱导的p-IκB蛋白、TNF-α及IL-1βmR-NA表达。结论 DATS可通过抑制p38MAPK通路抑制IκB磷酸化及NF-κB活化,进而下调LPS诱导小鼠肺泡巨噬细胞TNF-α、IL-1βmRNA表达。
Aim To investigate the role of p38 mitogen activated protein kinase （MAPK） in the signal transduction mechanisms of diallyl trisulfide （DATS） inhibiting the tumor necrosis factor （TNF-α） and interleukin-1 β （ IL-1β ） expression induced by lipopo-lysaccharide （LPS） in mouse alveolar macrophages cell line MH-S. Methods MH-S ceils were cultured in vitro, and treated with DATS in the presence or absence of LPS for different time. The expression of phospho- p 3 8 and p38 were assayed by Western blot.MH-S cells were incubated with specific p38 inhibitor SB203580 in the presence or absence of LPS. The expression of TNF-α mRNA and IL-1β mRNA were detected with reverse transcription-PCR （RT-PCR）. The expression of phospho-IKB and IKB were assayed by Western blot. Results The expression of the phospho- p38 expression in MH-S increased markedly under the stimulation of LPS in a time-dependent manner. Pretreatment with DATS（0. 1, 0. 5, 2. 5, 5.0 mg · L^-1 ） for 30min prior to LPS activation resulted in an obvious reduction of phospho-p38 expression in a dose-dependent manner. DATS alone did not influence the phos-pho-p38 expression. SB203580 dose-dependendy inhibited LPS-induced TNF-α mRNA, IL-1β mRNA expression and phospho-IKB expression. Conclusions DATS downregulates TNF-α and IL-1β mRNA expression in LPS-stimulated MH-S by inhibiting the p38MAPK pathway and the subsequent phosphorylation of κB and NF-κB activation.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2012年第9期1303-1307,共5页
Chinese Pharmacological Bulletin
基金
河北省科技攻关资助项目(No 05276101D-65)
关键词
二烯丙基三硫
脂多糖
肺泡巨噬细胞
P38丝裂原活化蛋白激酶
核因子-κB
肿瘤坏死因子α
白细胞介素1Β
diallyl trisulfide （DATS）
lipopolysaccha-ride（LPS）
alveolar macrophages
p38 mitogen activated protein kinase （p38 MAPK）
nuclear factor-κB（NF-κB）
tumor necrosis factor-α（TNF-α）
interleu-kin 1 β（IL-1β）
