论文部分内容阅读
目的研究哮喘气道重塑大鼠尾加压素Ⅱ(UⅡ)和转化生长因子(TGF)-β1的表达变化,探讨UⅡ对哮喘大鼠气道平滑肌细胞(ASMC)中TGF-β1表达的调控作用。方法以卵清白蛋白(OVA)致敏与激发建立哮喘大鼠气道重塑模型,16只雄性SD大鼠随机分为对照组和哮喘组,每组8只。分别采用免疫组织化学法(IHC)和反转录-聚合酶链反应(RT-PCR)法测定UⅡ和TGF-β1的蛋白和mRNA的表达。体外培养哮喘大鼠ASMC,分为对照组和UⅡ刺激组,其中UⅡ刺激组根据UⅡ干预时间的不同分为:4、8、12、24和48h组;按加入UⅡ浓度的不同分为:0.4、4、10、40和400nmol/L组。酶联吸附试验(ELISA)检测细胞培养液中TGF-β1蛋白含量,实时定量PCR(real-time PCR)检测ASMC中TGF-β1mRNA表达变化。结果哮喘组肺组织UⅡ蛋白、TGF-β1蛋白表达较对照组均明显升高,具有显著性统计学意义(P<0.01);哮喘组肺组织中UⅡmRNA和TGF-β1mRNA含量与对照组比较均明显增加,具有显著性统计学意义(P<0.01)。体外实验显示:UⅡ不同时间点促哮喘大鼠ASMC中,TGF-β1 mRNA的表达4h开始上升,24h达到高峰;TGF-β1蛋白表达于12h开始增加,24h达到高峰。UⅡ不同浓度促哮喘大鼠ASMC中,TGF-β1mRNA和蛋白表达均于4nmol/L开始上升,40nmol/L达到高峰。结论哮喘大鼠肺组织中UⅡ和TGF-β1呈高表达;UⅡ对哮喘大鼠ASMC中TGF-β1的表达可能存在调控作用,且呈一定时间-浓度依赖性。
Objective To investigate the changes of urotensin Ⅱ (UⅡ) and transforming growth factor-β1 (TGF-β1) expression in airway remodeling of asthmatic rats and to explore the regulation of UⅡ on the expression of TGF-β1 in airway smooth muscle cells (ASMC) of asthmatic rats effect. Methods Airway remodeling was induced by ovalbumin (OVA) sensitization and challenge in asthmatic rats. Sixteen male SD rats were randomly divided into control group and asthma group, with 8 rats in each group. Immunohistochemistry (IHC) and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the protein and mRNA expression of UⅡ and TGF-β1. The ASMCs of asthmatic rats were cultured in vitro and divided into control group and UⅡstimulation group, the UⅡstimulation group was divided into 4, 8, 12, 24 and 48h groups according to the time of UII intervention; , 4, 10, 40 and 400 nmol / L groups. The content of TGF-β1 in cell culture medium was detected by enzyme linked immunosorbent assay (ELISA), and the expression of TGF-β1 mRNA in ASMC was detected by real-time PCR. Results The expression of UⅡprotein and TGF-β1 protein in asthma group were significantly higher than those in control group (P <0.01). The levels of UⅡmRNA and TGF-β1 mRNA in asthmatic group were significantly higher than those in control group Increase, with significant statistical significance (P <0.01). In vitro, the expression of TGF-β1 mRNA increased at 4h and peaked at 24h, and the expression of TGF-β1 increased at 12h and peaked at 24h. In UMC with different concentrations of UⅡ, the expression of TGF-β1 mRNA and protein in asthmatic rats increased from 4nmol / L to 40nmol / L. Conclusion The expression of UⅡ and TGF-β1 in lung tissue of asthmatic rats is highly expressed. UⅡ may regulate the expression of TGF-β1 in ASMC of asthmatic rats in a time-concentration dependent manner.