基质金属蛋白酶9在哮喘大鼠中性粒细胞中的表达

Expressions of matrix metalloproteinase-9 in blood polymorphonuclear leucocyte of asthmatic rats

目的:观察基质金属蛋白酶9(MMP-9)在哮喘大鼠血中性粒细胞(PMN)中的表达,探讨PMN能否通过合成MMP-9参与哮喘的发病。方法:健康雄性SD大鼠30只,随机分成哮喘组(A组)、布地奈德治疗组(B组)和正常对照组(C组),用卵蛋白(OVA)致敏和激发的方法建立哮喘模型。分离纯化血PMN,免疫细胞化学法检测MMP-9的表达水平,ELISA法检测血清基质金属蛋白酶抑制剂-1(TIMP-1)的浓度。结果:A组(0.196±0.011,OD值)PMNMMP-9的表达水平显著高于C组(0.100±0.010,OD值)(P<0.01),B组(0.135±0.008,OD值)PMNMMP-9的表达水平显著低于A组但高于C组(均P<0.01)。A组血清TIMP-1为(34.96±4.54)ng/mL,表达水平显著高于C组的(26.14±3.43)ng/mL(P<0.01),B组血清TIMP-1的表达水平显著低于A组但高于C组(P<0.05或P<0.01)。MMP-9与TIMP-1的表达水平呈显著正相关(n=29,r=0.713,P<0.01)。结论:哮喘大鼠PMN合成MMP-9的功能增加,PMN可能部分通过增加合成MMP-9参与哮喘的发病,这种功能可以部分被布地奈德所抑制。

Objective： To observe the expressions of matrix metalloproteinase-9 （MMP-9） in blood poIymorphonuclear leukocyte （PMN） of asthmatic rats. And to approach whether PMN could participate in asthma pathogenesis through synthesizing MMP-9. Methods： Thirty rats were randomly divided into three groups on average, including asthmatic group （group A）, budesonide treated group （group B） and control group （group C）. Blood PMN were isolated and purified. The expressions of MMP-9 were detected with immunocytochemical method in blood PMN. The concentration of matrix metallo-proteinase inhibitor- 1 （TIMP-1） was detected with ELISA. Results ： The level of MMP-9 in group A （0.196 ± 0.011, Optical density） were significantly higher than those in group C （0.100±0.010）（P〈0.01）. Levels of MMP-9 in group B （0.135±0.008） were significantly lower than those in group A, but higher than those in group C （all P〈0.01）. Levels of TIMF-1 in group A （34.96＋_4.54 ng/mL） were significantly higher than those in group C （26.14 ± 3.43 ng/mL）（P〈0.01）. Levels of TIMP-1 in group B （29.58 ± 2.41 ng/ mL）were significantly lower than those in group A, but higher than those in group C （P〈0.05 or P 〈0.01）. There was significant positive correlation between the level of MMP-9 and TIMP-1（n=29,r=0.713, P〈0.01）. Conclusion： Levels of MMP-9 synthesized by PMN increase in asthma rats. PMN may be implicated with asthma pathogenesis via increasing synthesis of MMP-9, but this function of PMN can be partly inhibited by budesonide.