附子理中汤联合骨髓间充质干细胞调控变应性鼻炎大鼠鼻黏膜T-bet和GATA3 mRNA表达的研究

Aconite root decoction for regulating the function of middle-warmer combined with bone marrow mesenchymal stem cells regulates expressions of T-bet and GATA3 mRNA Expression in nasal mucosa of rats with allergic rhinitis

目的研究附子理中汤联合骨髓间充质干细胞(MSCs)对变应性鼻炎大鼠鼻黏膜T盒转录因子(T-bet)、锌指蛋白3(GATA3)mRNA表达的影响。方法将40只雄性SD大鼠随机分为5组,每组8只。对照组大鼠不造模、不干预;模型组、MSCs组、附子理中汤组、附子理中汤+MSCs组大鼠均进行变应性鼻炎造模,其中模型组造模后不治疗,MSCs组造模后给予骨髓MSCs尾静脉注射1次,附子理中汤组造模后给予附子理中汤连续灌胃14 d,附子理中汤+MSCs组造模后给予骨髓MSCs尾静脉注射1次和附子理中汤连续灌胃14 d。比较各组大鼠干预前后的行为学积分;干预结束后ELISA法检测鼻腔灌洗液中白细胞介素-4(IL-4)、白细胞介素-13(IL-13)、干扰素-γ(IFN-γ)水平,qRT-PCR法检测鼻黏膜组织中T-bet和GATA3 mRNA表达情况。结果MSCs组、附子理中汤组和附子理中汤+MSCs组大鼠干预后行为学积分均明显低于干预前及模型组(P均<0.05),且附子理中汤+MSCs组大鼠干预后行为学积分均明显低于MSCs组和附子理中汤组(P均<0.05)。与对照组比较,模型组大鼠鼻腔灌洗液中IL-4、IL-13水平和鼻黏膜组织中GATA3 mRNA相对表达量均明显升高(P均<0.05),鼻腔灌洗液中IFN-γ水平和鼻黏膜组织中T-bet mRNA相对表达量均明显降低(P均<0.05)。与模型组比较,MSCs组、附子理中汤组和附子理中汤+MSCs组大鼠鼻腔灌洗液中IL-4、IL-13水平和鼻黏膜组织中GATA3 mRNA相对表达量均明显降低(P均<0.05),附子理中汤组和附子理中汤+MSCs组大鼠鼻腔灌洗液中IFN-γ水平和鼻黏膜组织中T-bet mRNA相对表达量均明显升高(P均<0.05),MSCs组大鼠鼻腔灌洗液中IFN-γ水平和鼻黏膜组织中T-bet mRNA相对表达量均无明显变化(P均>0.05);且附子理中汤+MSCs组大鼠鼻腔灌洗液中IL-4、IL-13水平和鼻黏膜组织中GATA3 mRNA相对表达量均明显低于MSCs组(P均<0.05),鼻腔灌洗液中IFN-γ水平和鼻黏膜组织中T-bet mRNA相对表达量均明显高于MSCs组(P均<0.05)。结论附子理中汤联合骨髓MSCs能上调T-bet mRNA和下调GATA3 mRNA表达,进而调节Th1/Th2细胞的平衡,抑制炎症反应,改善变应性鼻炎症状。

Objective It is to study the synergistic effect of Aconite root decoction for regulating the function of middle-warmer(ARDRFM)combined with bone marrow mesenchymal stem cells(MSCs)on the mRNA expressions of T-box transcription factor(T-bet)and GATA-binding protein 3(GATA3)in nasal mucosa of rats with allergic rhinitis.Methods Forty male SD rats were randomly divided into 5 groups,with 8 rats in each group.The rats in the control group were not modeled and treated;the rats in the model group,MSCs group,ARDRFM group and ARDRFM+MSCs group were used to establish models of allergic rhinitis,the rat of the model group was not treated after modeling,those of the MSCs group were given one tail-vein injection of MSCs after modeling,those of the ARDRFM group were treated with ARDRFM by gavage for 14 days after modeling,those of the ARDRFM+MSCs group were treated with ARDRFM combined with MSCs.The behavioral scores of rats in each group were compared before and after treatment.At the end of intervention,the levels of interleukin-4(IL-4),interleukin-13(IL-13),and interferon-gamma(IFN-γ)in nasal lavage fluid were detected by ELISA,and the mRNA expressions of T-bet and GATA3 in nasal mucosa tissues were detected by qRT-PCR.Results The behavioral scores of rats in the MSCs group,ARDRFM group and ARDRFM+MSCs group were significantly lower after treatment than those before treatment and in the model group(all P<0.05),and the behavioral scores of rats in the ARDRFM+MSCs group were significantly lower than those in the MSCs group and ARDRFM group after treatment(all P<0.05).Compared with the control group,the levels of IL-4 and IL-13 in nasal lavage fluid and the relative expressions of GATA3 mRNA in nasal mucosal tissues of rats in the model group were significantly increased(all P<0.05),and the levels of IFN-γin nasal lavage fluid and the relative expression of T-bet mRNA in nasal mucosal tissues were significantly decreased(all P<0.05).Compared with the model group,the levels of IL-4 and IL-13 in nasal lavage fluid and the relative expression of GATA3 mRNA in nasal mucous tissues of rats in the MSCs group,ARDRFM group and ARDRFM+MSCs group were significantly decreased(all P<0.05),and the levels of IFN-γin nasal lavage fluid and the relative expression of T-bet mRNA in nasal mucous tissues of rats in the ARDRFM group and ARDRFM+MSCs group were all significantly increased(all P<0.05),while the level of IFN-γin nasal lavage fluid and the relative expression of T-bet mRNA in nasal mucous tissue of rats in the MSCs group were not changed significantly(both P>0.05);the levels of IL-4 and IL-13 in nasal lavage fluid and the relative expression of GATA3 mRNA in nasal mucous tissues of rats in the ARDRFM+MSCs group were significantly lower than those in the MSCs group(all P<0.05),and the level of IFN-γin nasal lavage fluid and the relative expression of T-bet mRNA in nasal mucosa tissue were significantly higher than those in the MSCs group(all P<0.05).Conclusion Aconite root decoction for regulating the function of middle-warmer combined with bone marrow mesenchymal stem cells can up-regulate expression of T-bet mRNA and down-regulate expression of GATA3 mRNA,thus to regulate the balance of Th1/Th2 cells,inhibit inflammatory response,and improve symptoms of allergic rhinitis.