CAJ | 학술논문

目的：气道重塑是支气管哮喘(简称哮喘)的主要病理特征之一,它与气道平滑肌细胞迁移和增殖功能密切相关,前列腺素 E 受体(E-prostanoid,EP)对之有调控功能。本研究通过大鼠哮喘模型构建,评价了哮喘状态下气道平滑肌细胞上 EP 改变情况,为开发 EP 类药物治疗哮喘气道重塑方法提供理论依据。方法20只 SD 清洁级大鼠,随机分为卵白蛋白激发哮喘组和对照组,28 d 后处死,进行组织学检查,分离培养气道平滑肌细胞,用荧光定量 PCR 测定。结果大鼠哮喘模型经组织病理学检查符合哮喘气道重塑现象,气道平滑肌细胞 RNA 纯度 A260/280处于1.8~2.0之间,用 OligodTadaptor 作引物进行逆转录反应,以特异 miRNAs 引物为正向引物,以共同的与 adaptor 配对的通用引物为反向引物,进行实时荧光定量 PCR 扩增。用 GAPDH 作为内参。扩增反应体系为 miRNAQ-PCR 诊断试剂盒。EP1~4相对表达量(2-△△Ct )在对照组和哮喘组分别为(EP1：4.35±0.18,6.55±1.21；EP2：1.35±0.12,0.24±1.06；EP3：4.59±1.14,5.89±1.74；EP4：2.89±1.85,1.69±0.44)哮喘组 EP2/4显著下降,而EP1显著增高(P <0.01)。结论大鼠哮喘模型气道平滑肌细胞上 EP2/4减少,和 EP1表达增加,这可能是哮喘气道重塑的重要因素。
목적：기도중소시지기관효천(간칭효천)적주요병리특정지일,타여기도평활기세포천이화증식공능밀절상관,전렬선소 E 수체(E-prostanoid,EP)대지유조공공능。본연구통과대서효천모형구건,평개료효천상태하기도평활기세포상 EP 개변정황,위개발 EP 류약물치료효천기도중소방법제공이론의거。방법20지 SD 청길급대서,수궤분위란백단백격발효천조화대조조,28 d 후처사,진행조직학검사,분리배양기도평활기세포,용형광정량 PCR 측정。결과대서효천모형경조직병이학검사부합효천기도중소현상,기도평활기세포 RNA 순도 A260/280처우1.8~2.0지간,용 OligodTadaptor 작인물진행역전록반응,이특이 miRNAs 인물위정향인물,이공동적여 adaptor 배대적통용인물위반향인물,진행실시형광정량 PCR 확증。용 GAPDH 작위내삼。확증반응체계위 miRNAQ-PCR 진단시제합。EP1~4상대표체량(2-△△Ct )재대조조화효천조분별위(EP1：4.35±0.18,6.55±1.21；EP2：1.35±0.12,0.24±1.06；EP3：4.59±1.14,5.89±1.74；EP4：2.89±1.85,1.69±0.44)효천조 EP2/4현저하강,이EP1현저증고(P <0.01)。결론대서효천모형기도평활기세포상 EP2/4감소,화 EP1표체증가,저가능시효천기도중소적중요인소。
Objective Airway remodeling is a main pathological characteristic of bronchial asthma (asthma),and strongly associated with migration and proliferation of airway smooth muscle cells. E-prostanoid (EP)receptor can regulate airway remodeling.This study established a rat model of asthma and evaluated EP changes in airway smooth muscle cells under the asthmatic state so as to provide theoretical evidence for developing EP drugs to treat airway remodeling in asthma.Methods A total of 20 clean Sprague-Dawley rats were randomly assigned to asthma group and control group.Twenty-eight days later,they were sacrificed for histological examination.Airway smooth muscle cells were isolated,cultured and measured using quantitative fluorescent PCR.Results Histopathological examination revealed that rat models of asthma were in accordance with the manifestations of asthmatic airway remodeling.After reverse transcription,real-time quantitative fluorescent PCR was performed using miRNA Q-PCR diagnostic kit.GAPDH was considered the internal reference.Relative expressions of E-prostanoid 1-4 (EP1-4)(2-△△Ct )in the control and asthma groups were respectively as follows:EP1:4.35±0.18,6.55± 1.21;EP2:3.64±0.12,1.35±1.06;EP3:4.59±1.14,5.89 ±1.74;EP4:2.89 ±1.85,1.69 ±0.44.EP2/4 significantly decreased,but EP1 significantly increased in the asthma group (P <0.01).Conclusions These results suggested that the reduced EP2/4 and increased EP1 expressions in airway smooth muscle cells of rat models of asthma were probably important factors for asthmatic airway remodeling.