摘要
目的研究支气管哮喘豚鼠α防御素(RNP)的表达及P2Y2受体对其的影响。方法72只成年雄性豚鼠随机分为正常组(A组)和哮喘组(B组),以腹腔内注射联合雾化吸入卵白蛋白(OVA)复制哮喘模型。将A组及B组分为生理盐水组(A1/B1组)、ATP干预组(A2/B2组)、苏拉明干预组(A3/B3组),分别对豚鼠腹腔内注射生理盐水、ATP及苏拉明。检测豚鼠气道外周阻力(RL)、肺泡灌洗液(BALF)细胞总数及中性粒细胞(PMN)计数;HE染色显示炎性细胞浸润情况;ELISA法检测血浆RNP和白细胞介素8(IL-8)表达含量;免疫组化和RT-PCR方法检测肺组织中RNP蛋白和mRNA以及P2Y2 mRNA的表达情况。结果哮喘组及其干预组豚鼠RL上升幅度较A组及其干预组增加10%(P<0.05)。HE染色证实哮喘组豚鼠支气管管腔变窄,炎性细胞浸润。B1组BALF中细胞总数及PMN计数高于A1组和B3组,低于B2组(P<0.05);A组间比较,差异无统计学意义。B1组血浆和肺组织中RNP表达较A1组和B3组增高,较B2组表达减低(P<0.05);A1组较A2组表达减低,与A3组比较增高(P<0.05)。B1组血浆中IL-8表达较A1组和B3组增高,较B2组减低(P<0.05);A组及其干预组比较,无明显差别(P>0.05)。RNP主要表达于细支气管中,尤其PMN浸润部位。RNP mRNA表达结果与肺组织中RNP表达趋势一致。A1组P2Y2 mRNA较A2组表达降低,与A3组比较增高(P<0.05);B1组P2Y2 mRNA表达较B2组降低,与B3组比较增高(P<0.05)。直线相关分析显示A组及其干预组RNP与PMN、IL-8无明显相关性,RNP与P2Y2mRNA呈正相关;B组及其干预组RNP与PMN、IL-8、P2Y2 mRNA呈正相关。结论α防御素在支气管哮喘豚鼠中表达增高;豚鼠α防御素表达可能与P2Y2受体有关。
Objective To investigate the expression of RNP（alpha defensins） in guinea pigs with bronchial asthma and the influences of P2Y2 receptors on the expression.Methods Seventy-two adult male guinea pigs were randomly divided into a control group（group A） and an asthma group（group B）.The asthma model was established by sensitizing with ovalbumin injection intraperitoneally and challenging with ovalbumin inhalation.Then the A and B groups were divided into following subgroups,ie.the saline groups（A1/B1）,ATP groups（A2/B2）,and Suramin groups（A3/B3）,in which the animals were intervented with saline,ATP,and suramin respectively.Total pulmonary resistance（RL） was measured.Total and differential cell counts in bronchoalveolar lavage fluid（BALF） were measured.Pathological changes of lung were observed under light microscope using HE staining.The plasma levels of RNP and IL-8 were measured by ELISA.The protein and mRNA expressions of RNP and P2Y2 were detected by immunohistochemistry and RT-PCR.Results The RL increased significantly as much as 10% in group B compared with group A.Pathological study revealed that luminal narrowing of bronchus and inflammatory cells infiltration in the asthma group.The total cell and polymorphonuclear neutrophil（PMN） counts in BALF in group B1 were significantly higher than group A1 and B3,but lower than group B2（P0.05）,while no significant difference among group A1,A2,or A3 was found.The RNP expression in plasma and lung tissue in group B1 was higher than group A1 and B3,but lower than group B2（P0.05）.The RNP expression in plasma and lung tissue was lower in group A1 than group A2,but higher than group A3（P0.05）.The IL-8 expression in plasma in group B1 was significantly higher than group A and B3,but lower than group B2（P0.05）,while no significant difference among group A1,A2,or A3 was found（P0.05）.RNP protein expressed predominantly in lung tissues,especially where the PMN infiltrated.The expression of RNP mRNA were consistent with the R
出处
《中国呼吸与危重监护杂志》
CAS
2011年第6期551-556,共6页
Chinese Journal of Respiratory and Critical Care Medicine
基金
湖南省自然科学基金(编号:09JJ5019)
