目的探讨应用抗CD54抗体保护脓毒症小鼠肺脏功能并改善其免疫功能状态的机制。方法24只C57/BL6小鼠按完全随机法分为4组:假手术组(Sham组)、盲肠结扎穿孔(cecal ligation and puncture,CLP)模型小鼠±生理盐水组(CLP组)、CLP±抗CD54抗体组(Anti-CD54组)和CLP±抗CD54抗体同型对照组(Isotype组),每组6只。观察各组小鼠肺组织病理形态学改变,肺组织炎性因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF)mRNA、白介素(intedeukin,IL)-10和IL-6的mRNA表达水平,胸腺、脾脏和肺组织髓过氧化物酶(myeloperoxidase,MPO)活性,肺组织湿重/干重比值,外周血和腹腔灌洗液细菌清除率,胸腺和脾脏细胞凋亡情况。结果苏木精-伊红(hematoxvlin-eosin,HE)染色显示应用Anti-CD54组小鼠与CLP组和Isotype组小鼠比较,肺组织的损伤程度减轻,表现为间隔轻度增宽、无明显出血,少量炎性细胞浸润;Anti.D54组肺组织TNF-α(3.93±0.82)和IL-10(2.83±0.55)的mRNA水平,均显著低于CLP组[(8.22±2.34)、(6.05±1.52)]和Isotype组[(8.54±1.75)、(5.56±1.33)](P〈0.05);Anti-CD54组胸腺[(45±10)U/mg]、脾脏[(39±8)U/mg]和肺组织[(64±12)U/mg]匀浆MPO活性(P〈0.05),均显著低于CLP组[(98±13)、(78±12)、(105_±10)U/mg]和Isotype组[(88±20)、(90±16)、(110±16)U/mg](P〈0.05);Anti-CD54组(4.03±0.18)肺组织湿重/干重比值,显著低于CLP组(4.95±0.18)和Isotype组(4.81±0.31)(P〈0.05);Anti-CD54组血液(6±2)及腹腔灌洗液(5±2)细菌清除率显著高于CLP组[(76±18)、(72833±7176)]和Isotype组[(69±18)、(66532±143006)](P〈0.001);脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(terminal deoxynueleotidyl transferase mediated nick end labeling,TUNEL)结果显示?
Objective To investigated whether the anti-CD54 antibody can protect the lung function and improve the immune function on septic mice. Methods Experimental sepsis was induced by cecal ligation and puncture (CLP). 24 C57BL/6 mice were randomly divided into 4 groups: Sham group, CLP group(CLP±saline), Anti-CD54 group (CLP+Anti-CD54 antibody ) and Isotype group (CLP +isotype antibody). Pathological morphology changes of lung tissue were observed. Wet/dry ratios and myekoperoxidase(MPO) activities of lung tissue were measured. The expression of tumor necrosis factor-α(TNF-α), interleukin(IL) -10 and 1L-6 mRNA of lung tissue were detected by reverse transcriptive polymerase chain reaction (RT-PCR). The blood and peritoneal cavity lavage bacterial burden were assayed. The spleen and thymus were used to evaluate apoptotic level and their MPO activities were also measured. Results Compared with the CLP group and Isotype group, hematoxylin-eosin (HE) staining showed that the histological damages occurred in Anti-CD54 group were lower and expressed as interval slightly widened, no significant bleeding and a small amount of inflammatory cell infiltration. The RT-PCR results showed that the expression level of TNF-α(3.93±0.82) and IL-10 (2.83±0.55) mRNA in Anti-CD54 group were obviously lower than that of CLP group [ (8.22±2.34), (6.05±1.52)] and Isotype group [ (8.54±1.75), (5.56±1.33) ](P〈0.05). The MPO level of thymus[ (45±10) U/mg], spleen[ (39±8) U/mg] and lung [ (64±12) U/mg] tissue in Anti-CD54 group were lower than that of CLP group[ (98-±13), (78±12), (105±10) U/rag]and Isotype group[ (88±20), (90±16),( 110 ± 16) U/mg ] (P〈0.05). After treated with Anti -CD54 antibody (4.03 ±0.18), CLP mice lung showed lower wet -to -dry ratio compared with CLP group (4.95±0.18) and Isotype group(4.81±0.31 )(P〈0.05 ). Compared to the CLP group[ (76±18), (72 833±7 176) ] a