哮喘豚鼠模型血淋巴细胞及嗜酸细胞与肺微血管内皮细胞粘附性的研究

ADHESION OF LYMPHOCYTES AND EOSINOPHILS IN PERIPHERAL BLOOD OF ASTHMA GUINEA PIG MODEL AND LUNG MICROVASCULAR ENDOTHELIAL CELLS

观察哮喘豚鼠模型嗜酸细胞和淋巴细胞与肺微血管内皮细胞的粘附率 ;探讨肺微血管内皮细胞在哮喘豚鼠模型血清孵育下的功能变化及炎症细胞与其粘附的分子基础。设豚鼠哮喘模型及对照组观察分离的外周血淋巴细胞和嗜酸细胞与内皮细胞的粘附率。RT PCR法测定内皮细胞VCAM 1及eotaxinmRNA的表达强度 ;EMSA法测定NF κB和AP 1的DNA结合活性。与对照组血清孵育相比 ,肺微血管内皮细胞在模型组血清刺激下 :①与两组外周血淋巴细胞及仅与模型组嗜酸细胞的粘附率显著升高 (P <0 0 1) ;②VCAM 1和eotaxinmRNA表达量显著增多 (P <0 0 1或P <0 0 5 ) ,NF κB和AP 1的DNA结合活性也显著升高 (P <0 0 1)。肺微血管内皮细胞可被哮喘模型血清激活 ,使粘附分子、趋化因子及核转录因子的合成和分泌显著增多 ,从而显著提高其与炎症细胞的粘附率。嗜酸细胞的粘附需其自身及内皮细胞两者激活。肺微血管内皮细胞活化及NF kB和AP 1的DNA结合活性增高在哮喘发病中可能起重要作用。

To explore the adhesion of peripheral blood lymphocytes and eosinophils and lung microvascular endothelial cells (LMEC) in asthma. Guinea pigs were divided into asthma model group and control group; the adhesion rates of isolated peripheral lymphocytes, eosinophils and the cultured LMEC monolayers were assayed.The mRNA expressions of VCAM 1 and eotaxin in LMEC were determined by RT PCR.The DNA binding activities of NF κB and AP 1 of LMEC were detected by electrophoretic mobility shifts assay (EMSA). Under the stimulation with serum of asthma group:①the adhesion rates of lymphocytes, activated eosinophils and LMEC were significantly elevated( P <0 01). ②The VCAM 1 and eotaxin of mRNA expressions and the DNA binding activities of NF κB and AP 1 in LMEC were significantly increased after stimulation with serum of asthma group( P <0 01 or P <0 05). LMEC could be activated by stimulation of serum of asthma group and could increase the secretion of adhesion molecules and chemotactors, thus elevating the adhesion rate of inflammatory cells.The increase in adhesion rate between eosinophils and LMEC is dependant on the activation of them both. The results suggested that activation of LMEC and elevation of NF κB and AP 1 binding activities may promote the inflammatory process in asthma.