【摘要】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小鼠按完全随机法随机分为四组，假手术组(Sham组)、盲肠结扎穿孔模型小鼠+生理盐水组(cecal ligation and puncture, CLP组)、CLP+抗CD54抗体组(Anti-CD54组)和CLP+抗CD54抗体同型对照组(Isotype组)。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, TNF-α) mRNA、白细胞介素-10(interleukin, IL-10)和白细胞介素-6(interleukin, IL-6)的mRNA表达水平，胸腺、脾脏和肺组织髓过氧化物酶活性，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HE染色显示应用Anti-CD54组小鼠与CLP组和Isotype组小鼠相比，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-CD54组肺组织TNF-α(3.93±0.82)和IL-10(2.83±1.33)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)、脾脏(39±8)和肺组织(64±12)匀浆MPO活性(P＜0.05)，均显著低于CLP组(98±13,78±12，105±10)和Isotype组(88±20,90±16,110±16)(P＜0.05)；Anti-CD54组(4.03±0.18)肺组织湿重/干重比值，显著低于CLP(4.95±0.18)组和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)；TUNEL结果显示：Anti-CD54组胸腺(55±13)和脾脏(45±17)组织凋亡细胞百分比数显著少于CLP组(127±30,130±25)和Isotype组(223±15，110±42) (P＜0.05)。结论 抗CD54抗体对脓毒症小鼠具有一定的治疗作用，可能与其改善小鼠肺功能状态，提高血液和腹腔清除率，抑制小鼠脾脏和胸腺凋亡有关。
【Abstract】 Objective To investigated wheather the anti-CD54 antibody can protect the lung function and improve the immune function. Methods Experimental sepsis was induced by cecal ligation and puncture (CLP). 24 C57BL/6 mice were randomly divided into sham group (Sham), CLP group (CLP), CLP+Anti-CD54 antibody intravenously group (Anti-CD54) and CLP+isotype antibody intravenously group (Isotype). To observe the pathological morphology changes, wet/dry ratios, myekoperoxidase (MPO) of lung tissue. Measure the lung tissue expression of TNF-α, IL-6 and IL-10mRNA by RT-PCR. The blood and peritoneal cavity lavage bacterial burden were detected. The spleen and thymus were used to measure apoptotic level by tunel and their MPO activities were also assayed. Results Under light microscope, the degree of histological damage occurred in anti-CD54 antibody group was the slightest. The RT-PCR results showed that the quantity of expression with TNF-а(3.93±0.82) and IL-10 (2.83±1.33) mRNA in anti-CD54 antibody group were obviously lower than 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), spleen (39±8) and lung (64±12) tissue in anti-CD54 antibody group were lower than CLP group (98±13,78±12，105±10) and Isotype group (88±20,90±16,110±16)(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±18，72833±7176) and Isotype group (69±18,66532±143006), the peritoneal cavity lavage (5±2) and blood (6±2) bacterial clearance were elevated in anti-CD54 antibody group (P<0.001).After treated with anti-CD54 antibody, the TUNEL assays of thymus (55±13) and spleen (45±17) in anti-CD54 antibody were less than CLP group (127±30,130±25) and Isotype group (223±15，110±42)(P<0.01). Conclusions Anti-CD54 antibody could ameliorate the lung and immune function via increasing bacterial clearance and decreasing the immune organs’apoptosis.
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