脂多糖结合蛋白多抗对大鼠肺泡巨噬细胞内毒素耐受性及Toll样受体4表达的影响  

Effects of polyclonal lipopolysaccharide binding protein antibody on the expression of Toll-like receptor 4 and tolerance to lipopolysaccharide in alveolar macrophages of rats

作　　者：张德明[1] 邓青南[2] 李永旺[1] 毛宝龄[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 [2]广州军区广州总医院老年呼吸内科,广州510010

出　　处：《第三军医大学学报》2006年第2期140-143,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金资助重点项目(39730210)~~

摘　　要：目的观察脂多糖结合蛋白(lipopolysaccharidebindingprotein,LBP)多抗对大鼠肺泡巨噬细胞(alveolarmac-rophage,ΦAM)内毒素(lipopolysaccharide,LPS)耐受性与Toll样受体4(Toll-likereceptor4,TLR4)表达的影响,研究二者之间的联系。方法将从32只Wistar雄性大鼠分离所得ΦAM,按随机数字法等分为正常对照组(A)、LPS单次刺激组(B)、LPS二次重复刺激组(C)和LPS二次重复刺激+LBP多抗组。运用ELISA和RT-PCR方法分别检测各组大鼠ΦAM分泌肿瘤坏死因子-α(TNF-α)及TLR4mRNA表达的变化,Westernblot检测TLR4蛋白表达的变化。结果大鼠ΦAMTNF-α分泌和TLR4mRNA表达及TLR4的蛋白表达水平,A组分别为:(0·45±0·01),(0·77±0·06),(51·03±1·19)μg/L,B组分别为:(0·76±0·03),(1·55±0·25),(122·72±1·75)μg/L,B组与A组比较显著增加,P<0·01;C组分别为:(0·49±0·05),(1·13±0·07),(85·35±0·68)μg/L,C组与B组比较,明显减少,P<0·05。D组分别为:(0·28±0·07),(0·78±0·17),(61·06±1·36)μg/L,D组与C、B组比较,明显减少(P<0·05,P<0·01)。结论LPS重复刺激可使大鼠ΦAM对LPS产生耐受性;LPS耐受性的产生与TLR4表达下降相关;LBP多抗通过降低TLR4表达,提高大鼠ΦAM对LPS耐受性,此乃LBP抗体可用于治疗LPS所致急性肺损伤的部分分子生物学理论基础。Objective To explore the effects of polyclonal lipopolysaccharide binding protein （LBP） antibody on the expression changes of Toll-like receptor 4 （ TLR4 ） and the tolerance to lipopolysaccharide （LPS） in alveolar macrophages（DAM） of rats. Methods The DAM obtained from 32 Wistar male rats were randomly divided into control group （A）, LPS single stimulating group （B）, LPS re-timulating group （C） and LPS restimulating plus polyclonal LBP antibody group （D）, TNF-α concentration and TLR4 mRNA expression were measured by ELISA and semi-quantitive reverse transcription-polymerase chain reaction （RT-PCR） respective- ly in DAM of each group. The expression of TLR4 protein was measured by Western blotting, Results The TNF-α concentration and TLR4 expression in the DAM stimulated with single dose LPS （ 100 ng/ml ） were highly increased compared with the control group, but significantly decreased after re-stimulated with two dose of LPS （ 100 ng/ml and 500 ng/ml） and the reduction was more overt in group D, Conclusion The tolerance to LPS in DAM can be induced by re-stimulation with LPS. The down-regulated expression of TLR4 is related to the tolerance to LPS. Poly-clonal LBP antibody can enhance ФAM tolerance to LPS by decreasing TLR4 expression and the antibody is presumably eligible on the basis of molecular biology for the treatment of acute lung injury induced by LPS.

关 键 词：脂多糖结合蛋白多抗 肺胞巨噬细胞 内毒素 耐受性 TOLL样受体4 

分 类 号：R322.35[医药卫生—人体解剖和组织胚胎学] R392.11[医药卫生—基础医学]