摘要
目的研究脂多糖结合蛋白(LBP)抑制肽对内毒素(LPS)诱导的人单核巨噬细胞株U937细胞表达CD14的影响。方法佛波脂(PMA)诱导U937成熟后分为5组:正常对照组、LPS组(100ng/mLLPS+100ng/mLrhLBP)、低剂量抑制肽组、中剂量抑制肽组及高剂量抑制肽组,后3组分别给予10、100和1000ng/mL抑制肽。用RTPCR和蛋白定量方法(Westernblot)测定CD14mRNA和蛋白的表达,ELISA测定培养上清液中肿瘤坏死因子α(TNFα)的含量。结果LBP抑制肽显著抑制LPS刺激后U937细胞CD14的mRNA与蛋白表达,同时也抑制了TNFα的分泌,较LPS组有显著差异。结论LBP抑制肽通过抑制由CD14介导的LPS信号跨膜转导和TNFα的分泌,对LPS所致疾病如脓毒血症、急性肺损伤可能有潜在的治疗作用。
Objective To investigate the effects of lipopolysaccharides (LPS) binding protein (LBP) inhibitory peptide on CD14 expression on U937 cells induced by LPS.Methods U937 cells were stimulated by Phorbol-myristate-acetate (PMA,100 ng/mL) and divided into five groups:control group,LPS group (100 ng/mL LPS plus 100 ng/mL rhLBP),low dose inhibitory peptide group,middle dose inhibitory peptide group,and high dose inhibitory peptide group (10 ng/mL,100 ng/mL,and 1 000 ng/mL,respectively).The mRNA and protein expressions of CD14 was determined by RT-PCR and western blot and the concentration of TNF-α released from U937 cells was detected by ELISA.Results CD14 mRNA and protein expressions in inhibitory peptide group were lower than those in LPS group and higher than those in control group.The concentration of TNF-α in inhibitory peptide group was lower than that in LPS group and higher than that in control group.Conclusion LBP inhibitory peptide might have a potential protective effect on LPS induced inflammatory disorders such as sepsis and acute lung injury through inhibiting the mRNA and protein expression of CD14 and reducing the release of TNF-α.
出处
《中国呼吸与危重监护杂志》
CAS
2005年第3期230-232,共3页
Chinese Journal of Respiratory and Critical Care Medicine
基金
国家自然科学基金资助(30170366)
