摘要
目的:从肺组织中基质金属蛋白酶-2(MMP-2)、转化生长因子β1(TGF-β1)的异常表达着手,探讨短发夹RNA(shRNA)介导结缔组织生长因子(CTGF)沉默对新生大鼠肺部血管重构和支气管肺发育不良(BPD)的影响及其机制。方法:将30只新生大鼠按随机数字表法分为空白对照组、博来霉素组、博来霉素+CTGF处理组,每组10只。大鼠自出生后第1天开始进行造模,空白对照组不造模,博来霉素组、博来霉素+CTGF处理组进行BPD-肺动脉高压(PH)造模,博来霉素+CTGF处理组在第7天给予CTGFshRNA慢病毒经滴鼻给药2μL,共1次,共1d。第14天采集标本,右肺常规石蜡包埋切片,用于HE染色。测量右心室肥厚指数[右心室(RV)/左心室(LV)+室间隔(VS)的质量],获取肺部组织HE染色后观察辐射状肺泡计数(RAC),计算管壁厚度占血管外径的百分比(WT%)及管壁面积占血管总面积的百分比(WA%)。Western Blot法检测CTGF、TGF-β1、MMP-2蛋白的表达水平,RT-PCR法检测CTGF、TGF-β1、MMP-2的mRNA表达。结果:与空白对照组相比,博来霉素组大鼠肺泡结构不完整,肺泡腔增大,肺小动脉管壁增厚,WT%、WA%、RV/(LV+S)增高(P<0.05),RAC降低(P<0.05),MMP-2、TGF-β1、CTGF蛋白及mRNA增高(P<0.05);与博来霉素组相比,博来霉素+CTGF处理组大鼠肺小动脉管壁增厚改善,肺泡结构存在,肺泡腔增大不明显,WT%、WA%、RV/(LV+S)降低(P<0.05),RAC增高(P<0.05),MMP-2、TGF-β1、CTGF蛋白及mRNA降低(P<0.05)。结论:新生大鼠BPD-PH模型存在CTGF、TGF-β1、MMP-2蛋白及mRNA高表达,下调CTGF可减轻BPD-PH动物模型的肺血管重塑、肺部病变的发生,可能与TGF-β1、CTGF、MMP-2有关。
Objective:Based on the abnormal expression of matrix metalloproteinase-2(MMP-2)and transforming growth factorβ1(TGF-β1)in lung tissue,to investigate the effects of short hairpin RNA(shRNA)mediated silencing of connective tissue growth factor(CTGF)on pulmonary vascular remodeling and bronchopulmonary dysplasia(BPD)in neonatal rats.Methods:Thirty newborn rats were divided into blank control group,bleomycin group and bleomycin+CTGF group by random number table method,with ten rats in each group.The rats were molded from the first day after birth.The blank control group was not modeled,and the bleomycin group and bleomycin+CTGF treatment group were modeled with PBD-pulmonary hypertension(PH).Bleomycin+CTGF treatment group was given 2μL of CTGF shRNA lentivirus by nasal drop on day 7,once for 1 day.Specimens were collected on the 14th day,and the paraffin embedded sections of the right lung were used for HE staining.Right ventricular hypertrophy index[mass of right ventricle(RV)/left ventricle(LV)+interventricular septum(VS)]was measured.After HE staining,radiative alveolar count(RAC)was observed,and the percentage of tube wall thickness to outer diameter of blood vessels(WT%)and the percentage of tube wall area to total blood vessels(WA%)were calculated.The expression levels of CTGF,TGF-β1 and MMP-2 proteins were detected by Western Blot,and the mRNA expressions of CTGF,TGF-β1 and MMP-2 were detected by RT-PCR.Results:Compared with blank control group,bleomycin group had incomplete alveolar structure,enlarged alveolar cavity,thickened pulmonary arteriole wall,increased WT%,WA%,RV/(LV+S)(P<0.05),decreased RAC(P<0.05),and increased MMP-2,TGF-β1,CTGF protein and mRNA(P<0.05);Compared with bleomycin group,bleomycin+CTGF treatment group improved pulmonary artery wall thickness,alveolar structure existed,and alveolar cavity enlargement was not obvious,WT%,WA%,RV/(LV+S)were decreased(P<0.05),RAC was increased(P<0.05),MMP-2,TGF-β1,CTGF protein and mRNA were decreased(P<0.05).Conclusion:The protein and mRNA expression of CTGF,TGF-β1 and MMP-2 were high in the neonatal rat BPD-PH model.Down-regulation of CTGF could reduce the pulmonary vascular remodeling and the occurrence of lung lesions in the BPD-PH animal model,which may be related to TGF-β1,CTGF and MMP-2.
作者
何中倩
黄海云
邱海蓉
李晓东
He Zhongqian;Huang Haiyun;Qiu Hairong;Li Xiaodong(Department of Neonatology,Union Shenzhen Hospital of Huazhong University of Science and Technology,Shenzhen 518052,Guangdong Province,China)
出处
《中外医药研究》
2023年第14期3-6,共4页
JOURNAL OF CHINESE AND FOREIGN MEDICINE AND PHARMACY RESEARCH
基金
2021年深圳市南山区科技计划项目(编号:NS087)。
关键词
支气管肺发育不良
管壁增厚
肺部组织
肺泡腔
博来霉素
小动脉
结缔组织生长因子
右肺
Bronchopulmonary dysplasia
Pulmonary vascular remodeling
Connective tissue growth factor
Transforming growth factorβ1
Matrix metalloproteinase-2
