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目的探讨A20重组蛋白对支气管哮喘小鼠气道重构及NF-κB信号通路的影响。方法 40只雄性清洁级Balb/c小鼠,随机数字表法分为4组,每组10只,分别为:生理盐水对照组;卵蛋白(OVA)哮喘组;A20重组蛋白治疗3 d组;A20重组蛋白治疗7 d组。在末次激发24 h后所有小鼠取左肺组织行苏木精-伊红(HE)染色及PAS染色。取右肺组织分别用酶联免疫吸附法(ELISA),RT-PCR和Western blote检测支气管肺泡灌洗液(BALF)中IL-4、IL-5、TNF-α及IFN-γ含量以及肺组织中结缔组织生长因子(Connective tissue growth factor,CTGF)、转化生长因子-β1(trailsforming growth factor-β1,TGF-β1)的mRNA表达和核转录因子(nuclear factor kappa B,NF-κB)的表达。结果哮喘模型组小鼠与对照组相比较BALF中炎症细胞计数、IL-4、IL-5、TNF-α水平增高,而IFN-γ水平降低;肺组织CTGF、TGF-β1的转录和表达水平以及NF-κB表达水平均显著高于对照组(P<0.01)。A20重组蛋白3 d和7 d干预组小鼠与哮喘模型组相比较BALF中炎症细胞计数、IL-4、IL-5、TNF-α水平,CTGF、TGF-β1的转录和表达水平,NF-κB表达水平均显著降低,而BALF中IFN-γ水平明显上升,具有显著差异(P<0.05),但2个不同治疗组之间上述各指标间差异无统计学意义(P>0.05)。结论 A20重组蛋白可抑制哮喘小鼠气道重构的发生,其机制有可能是通过抑制NF-κB/TGF-β1/CTGF信号通路而实现的。
Objective To investigate the effect of A20 recombinant protein on airway remodeling and NF-κB signaling in bronchial asthmatic mice. Methods Forty male Balb / c male mice were randomly divided into 4 groups (n = 10): saline control group, ovalbumin (OVA) asthma group, A20 recombinant protein 3 d group, A20 recombinant protein treatment 7 d group. All the mice were sacrificed at 24 h after the last challenge. The left lungs were stained with hematoxylin and eosin (HE) and PAS. The contents of IL-4, IL-5, TNF-α and IFN-γ in bronchoalveolar lavage fluid (BALF) were detected by enzyme linked immunosorbent assay (ELISA), RT- The expression of connective tissue growth factor (CTGF), transforming growth factor-β1 (TGF-β1) and nuclear factor kappa B (NF-κB) Results Compared with the control group, the number of inflammatory cells, IL-4, IL-5 and TNF-α levels in BALF and the level of IFN-γ in BALF of asthmatic model group were significantly decreased. The transcription and expression levels of CTGF and TGF- And NF-κB expression levels were significantly higher than the control group (P <0.01). The levels of IL-4, IL-5, TNF-α, CTGF, TGF-β1 transcription and expression of NF-κB in BALF were significantly increased in the 3-day and 7-day interventional A20 recombinant proteins compared with the asthma model group (P <0.05). However, there was no significant difference between the two groups (P> 0.05). Conclusion A20 recombinant protein can inhibit the airway remodeling in asthmatic mice, and its mechanism may be through the inhibition of NF-κB / TGF-β1 / CTGF signaling pathway.