CD14抑制肽的体外生物活性  

Biological activity of CD14 inhibitory peptide in vitro

作　　者：冯起甲[1] 徐智[1] 吴国明[1] 刘红艳 徐顺贵[1] 钱桂生[1] 

机构地区：[1]第三军医大学新桥医院全军呼吸内科研究所全军呼吸病研究重点实验室,重庆400037 [2]第五三四医院,洛阳471003

出　　处：《中国急救医学》2007年第8期716-719,共4页Chinese Journal of Critical Care Medicine

基　　金：国家自然科学基金(No30500230)

摘　　要：目的研究CD14抑制肽(CD14inhibitory peptide,CD14-IP)与CD14的结合活性及对内毒素(LPS)和脂多糖结合蛋白(LBP)诱导的人单核巨噬细胞株U937表达肿瘤坏死因子-α(TNF-α)的影响。方法采用酶联免疫吸附(ELISA)法进行CD14-IP与CD14的结合实验。U937细胞用佛波脂(PMA)诱导成熟后分为五组:正常对照组、LPS组(100ng/mLLPS+100ng/mLrhLBP)、高剂量多肽组、中剂量多肽组及低剂量多肽组,后三组分别给予10μg/mL、1.0μg/mL和0.1μg/mL的CD14-IP。用ELISA测定培养细胞上清TNF-α的含量,用RT-PCR测定U937细胞TNF-αmRNA的表达水平。结果CD14-IP有较强的与CD14结合的能力;CD14-IP组TNF-α和TNF-αmRNA水平均较LPS组低,较正常组高。结论CD14-IP能与CD14结合,并能降低培养细胞TNF-α和TNF-αmRNA的表达,具有抑制CD14生物活性的作用。Objective To investigate the binding efficacy of CD14 inhibitory peptide （ CD14 - IP） binding CD14 and the effect of CD14 - IP on TNF -α expression in U937 cells induced by LPS. Methods The binding test of CDI4 - IP binding to CD14 was carried out by ELISA. Then, U937 cells were stimulated by Phorbol myristate acetate （ PMA ） and divided into five groups： control group, LPS group （ 100 ng/mL LPS plus 100 ng/mL rhLBP） , high dose inhibitory peptide group, middle dose inhibitory peptide group and low dose inhibitory peptide group （ 10 μg/mL, 1.0 μg/mL , and 0.1 μg/mL , respectively） . The concentration of TNF-α released from cells was detected by ELISA. The mRNA of TNF-α in U937 cells was determined by RT - PCR. Results CD14 - IP has a high binding efficacy with CD14. The the concentration of TNF -α and level of TNF-α mRNA in inhibitory peptide group were significantly lower than that in LPS group, and were significantly higher than that in control group. Conclusion CD14 - IP has a high binding efficacy with CD14, and can reduce the level of TNF -α and TNF-α mRNA in U937 cells induced by LPS. It can inhibit biological activity of CD14.

关 键 词：CD14 多肽 脂多糖结合蛋白 脂多糖 TNF-Α 

分 类 号：R392[医药卫生—免疫学]