摘要
目的观察己酮可可碱(PTX)对内毒素(LPS)诱导大鼠心肌细胞间黏附分子1(ICAM 1)表达的影响,探讨PTX对心肌的保护作用及其机制。方法将培养的心肌细胞分为3组:空白对照组、LPS损伤组和药物保护组。空白对照组心肌细胞不予任何处理。LPS组心肌细胞中一部分用100μg/L LPS刺激2、4、6和8 h,另一部分用10、50和100μg/L LPS刺激6 h,采用酶联免疫吸附法(EL ISA)检测心肌细胞ICAM 1的表达,免疫细胞化学法检测核转录因子κB(NFκB)p65的表达,蛋白质免疫印迹法(W estern b lot)检测NFκB抑制蛋白α(IκBα)表达;并观察LPS处理前加入50、100、200 m g/L PTX对心肌细胞ICAM 1表达的影响。结果心肌细胞ICAM 1表达与LPS刺激呈时相剂量依赖方式;LPS刺激可迅速活化NFκB p65,于1h时达高峰;IκBα表达先降低后升高;加入PTX后大鼠心肌细胞ICAM 1、NFκB p65表达下降,IκBα升高。结论PTX可通过抑制ICAM 1上游NFκB途径对LPS造成的心肌损伤起保护作用。
Objective To investigate the effects of pentoxifylline （PTX） on the expression of intercellular adhesion molecule - 1 （ICAM - 1 ） of rat cardiac myocytes after lipopolysaccharide （LPS） challenge in vitro, and to evaluate the protective effect of PTX on cardiac myoeytes and its mechanism. Methods Cultured cardiac myocytes were randomly divided into three groups： control group, LPS group and PTX protection group. No drug was given in the control group. In LPS group, a part of cardiac myocytes were stimulated with 100/μg/L of LPS for 2, 4, 6, 8 hours, respectively, and the others were stimulated with 10, 50, 100μg/L LPS, respectively, for 6 hours. Expression of ICAM - 1, nuclear factor - κB （NF - κB） p65 and NF - κB inhibitor -α（IκB -α） in cardiac myocytes was respectively measured with enzymelinked immunoadsorbent assay （ELISA）, immunohistochemistry, and Western blot. The expression of ICAM - 1 in cardiac myocyte was also determined after the addition of 50, 100, 200 mg/L of PTX before LPS challenge. Results The intensity of expression of ICAM - 1 was highly dependent on the stimulation of LPS in dose depending and time -depending manner. The stimulation quickly resulted in activation of NF - κB p65 to translocate into nuclei. This process peaked at 1 hours and then declined. The expression of IκB-α was depressed at first, then elevated. With the addition of PTX, the expression of ICAM - 1 and NF κB p65 was depressed, and the expression of IκB -α was elevated. Conclusion PTX may have a protective effect on the cardiac myocytes against NF - κB, which regulates the production of ICAM - 1.
出处
《中国危重病急救医学》
CAS
CSCD
北大核心
2006年第2期109-112,I0001,共5页
Chinese Critical Care Medicine
基金
江苏省南通市社会发展科技计划项目(S30011)
关键词
内毒素
心肌细胞
细胞间黏附分子-1
核转录因子-KB
己酮可可碱
lipopolysaccharide
cardiac myocyte； factor- κB
pentoxifylline LPS injury through inhibiting the pathway of intercellular adhesion molecule - 1
nuclear
