CD14的表达及其在库普佛细胞激活与所致组织损伤中的意义(英文)

CD14 expression and its significance in activation of Kupffer cells and its induced injury

背景:如何减轻甚至消除脂多糖(LPS)引起的全身炎症反应是目前研究的热点;库普佛细胞(KC)的激活与LPS的这种作用密切相关,但关于其具体的机制尚有待于进一步研究。目的:探讨LPS对KCCD14表达的影响及CD14在LPS激活KC中的意义。设计:完全随机前后对照研究。地点和对象:在第三军医大学西南医院病理学研究所完成。大鼠KC来源于Wistar大鼠,购自第三军医大学实验动物中心。干预:在分离培养大鼠KC的基础上,作者应用LPS直接或LPS刺激KC后产生的介质刺激新培养的KC,或在血清存在的情况下加入抗CD14单抗或在无血清的情况下单独加入LPS等措施刺激KC细胞。主要观察指标:各种条件下CD14mRNA的表达及其蛋白合成的变化、培养KC中上清中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和一氧化氮浓度。结果:①CD14mRNA表达及其蛋白合成在正常KC中极弱,分别为0.035和19.91±2.89,浓度为10mg/L的LPS分别为0.73和676.79±24.95,其量与LPS浓度呈剂量依赖性相关。浓度为10μg/L的LPS刺激后KC中CD14mRNA表达及其蛋白合成在30min即明显增强,分别为0.56和548.86±40.43,至6h达高峰(0.54和673.91±35.08)。②在血清存在时加入抗CD14单抗或在无血清时单独加入LPS,可明显降低KCTNF-α,IL-6和一氧化氮的释放。而后者如果同时加入LBP,则可?

BACKGROUND:It is a hotspot of recent researches to reduce and even eliminate the general inflammation induced by lipopolysaccharide(LPS).The activation of Kupffer cell(KC)closely relates with the reaction of LPS;however,its detailed mechanism still requires further investigations. OBJECTIVE:To investigate the impact of LPS in the expression of KC CD14 as well as the significance of CD14 in the activation of KC by LPS DESIGN:A Completely randomized auto controlled trial. SETTING and PARTICIPANTS:Study was conducted in the Research Institute of Pathology Southwest Hospital,Third Military Medical University of Chinese PLA.Rat KC was originated from Wistar rats obtained from the experimental animal centre of the Third Military Medical University. INTERVENTION:On the basis of KC separation and culture,the author used the mediators produced directly by LPS or after LPS stimulating KC to stimulate newly cultured KC,or added anti CD14 mono antibody with the existence of serum or added LPS solely without the existence of serum to stimulate KC. MAIN OUTCOME MEASURES:The expression of CD14 mRNA,the alteration of proteinic synthesis,tumor necrosis factor α(TNFα),interleukin 6 (IL 6) and nitric oxide (NO) concentrations in the supernatant of KC culture under all kinds of conditions. RESULTS:① The expression of CD14 mRNA and the synthesis of protein were very weak in normal KC,which were 0.035 and 19.91±2.89 respectively.The expression of CD14 mRNA and the synthesis of protein in 10 mg/L of LPS were 0.73 and 676.79±24.95 respectively,which had dose dependence with LPS concentration. The expression of CD14 mRNA and the synthesis of protein in the KC stimulated by 10 μg/L of LPS significantly enhanced at 30 minutes,which were 0.56 and 548.86±40.43 respectively and reached its peak at 6 hours of 0.54 and 673.91±35.08 respectively.② It could significantly reduce the release of TNFα,IL 6 and NO from KC by adding CD14 mono antibody with the existence of serum or adding LPS solely without the existence of serum.If LBP were added simultaneously in the latter one, the concentrations of TNFα,IL 6 and NO would be significantly increased in KC culture. CONCLUSION:①The active mediator produced by LPS and after it stimulating KC closely relates with the expression of CD14 mRNA and its proteinic synthesis.It could be presumed that the enhancement of CD14 expression at 1-3 hours in the study might be mainly induced by LPS and the following further enhancement of CD14 expression might closely related with the cell factors released by KC.②The activation of KC by low concentration LPS is CD14 dependent.