酪氨酸激酶受体RON在慢性阻塞性肺疾病大鼠肺内表达的研究

The expression of receptor tyrosine kinase RON in lung tissue of rats with chronic obstructive pulmonary disease

目的:探讨酪氨酸激酶受体RON在慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)大鼠气道炎症中的作用。方法:单纯熏香烟法建立COPD大鼠模型;支气管肺泡灌洗计数支气管肺泡灌洗液(BALF)中细胞总数与肺泡巨噬细胞(alveolar macrophage,AM)数,培养正常和COPD大鼠AM,采用逆转录-聚合酶链式反应法检测大鼠肺组织中酪氨酸激酶受体RON mRNA的表达情况,免疫组化法观察大鼠气道和离体培养的AM中RON蛋白的表达水平,图像分析系统测定肺平均内衬间隔(MLI)、平均肺泡数(MAN)和肺泡腔面积与总面积比(PAA)。结果:COPD组MLI、PAA较正常对照组明显增高[MLI:(111.451±49.334)×10-6m vs(44.803±10.624)×10-6m;PAA:(79.653±7.594)%vs(48.352±13.063)%],而MAN则明显低于正常对照组[(81.621±20.394)×106/m2vs(170.098±42.398)×106/m2],差异均有统计学意义(P<0.01)。COPD组BALF中细胞总数和AM计数均明显高于正常对照组[细胞总数:(6.029±0.420)×108/L vs(1.463±0.0692)×108/L;AM数:(5.752±0.571)×108/L vs(1.387±0.105)×108/L,P<0.01)]。COPD组大鼠肺组织RON mRNA表达较正常对照组明显上调[(0.892±0.088)vs(0.353±0.080),P<0.01],COPD组大鼠气道上皮及AM中RON蛋白水平也较正常对照组显著增加[气道RON蛋白:(0.171±0.027)vs(0.073±0.009);AM RON蛋白:(0.310±0.101)vs(0.110±0.006),P<0.01]。结论:RON与COPD气道炎症调节密切相关。

Objective： To explore the role of receptor tyroslne kinase R ON in the airway inflammation of chronic obstructive pulmona-ry disease （COPD） in rats. Methods：A rat COPD model was established by exposing the rats to cigarette smoke daily for three mouth. Total cell counts and alveolar macrophage （AM） counts in bronchoalveolar lavage fluid （BALF） were examined. Rat AMs from the control group and COPD group were cultured. The expression of RON mRNA in lung tissue of rats was assessed by reverse transcription- polymerase chain reaction （RT-PCR）. The levels of RON protein in the airway of rats and AM cultured in vitro were observed by immuno -histochemistry. Results：The MLI and PAA in COPD group were higher than those of the control group while MAN was juston the contrary （P 〈0. 01 ）. The total cell counts and AM counts in BALF of the COPD group were significantly higher than those of control group （P 〈0. 01 ）. Compared to the control group,the levels of RON mRNA in lung tissue from COPD rats were up-regulated significantly （P 〈0. 01 ）. The levels of RON protein in the COPD rats airway and AM were higher than those of control group signifi-eanfly （P 〈0. 01 ）. Conclusion ：There is a close correlation between RON and airway inflammatory mechanism of COPD.