二陈汤加味对慢性阻塞性肺疾病大鼠肺组织GATA3,T-bet mRNA表达的影响

Effect of Modified Erchentang on GATA3 and T-bet mRNA Expressions in Lung Tissue of Chronic Obstructive Pulmonary Disease Rats

目的:观察二陈汤加味对慢性阻塞性肺疾病(COPD)大鼠肺组织锌指蛋白3(GATA3),T盒转录因子(T-bet)的影响。方法:将70只SD大鼠随机分为7组,每组10只,分别为正常组,模型组,二陈汤加味低、中、高剂量组(5,10,20 g·kg-1),消咳喘组(5 g·kg-1),二陈汤组。以烟熏合并脂多糖(LPS)气管滴注的方法制备COPD大鼠模型。成功造模后,治疗组灌胃给药,正常组及模型组灌胃等体积生理盐水。酶联免疫吸附测定(ELISA)测定大鼠血清中白细胞介素-10(IL-10),白细胞介素-12(IL-12)的含量。实时荧光定量聚合酶链式反应(Real-time PCR)检测GATA3,T-bet mRNA表达,免疫组化(IHC)检测其肺组织GATA3,T-bet的蛋白表达。结果:与正常组比较,模型组大鼠血清中IL-10水平显著降低,IL-12显著升高(P <0. 01);肺组织GATA3的蛋白和mRNA表达显著降低,T-bet的表达显著升高(P <0. 01)。与模型组比较,各治疗组血清中IL-10水平均有不同程度的升高,IL-12则不同程度的降低(P <0. 05)。各治疗组大鼠肺组织GATA3 mRNA和蛋白表达均明显增多,T-bet则受到了抑制(P <0. 05)。结论:二陈汤加味可能是通过降低IL-12,增加IL-10的含量,并抑制T-bet,兴奋GATA3的蛋白和mRNA表达,来减轻COPD大鼠肺组织炎症,改善肺功能的,并阻止COPD的免疫紊乱。

Objective:To observe the effect of modified Erchentang on GATA-binding protein-3(GATA3)and T-box expressed in T cells(T-bet)in lung tissue of rats with chronic obstructive pulmonary disease(COPD).Method:Seventy SD rats were randomly divided into seven groups,namely normal group,model group,low,medium and high-dose modified Erchentang group(5,10,20 g·kg-1),Xiaokechuan group(5 g·kg-1)and Erchentang group(5 g·kg-1),with 10 in each group.The rat model of COPD was established by smoking combined with intratracheal dripping of lipopolysaccharide(LPS).After successful modeling,the treatment group was given intragastric administration,and the normal group and the model group were given intragastric administration of equal volume of saline.Enzyme-linked immunosorbent assay(ELISA)was used to determine the concentrations of interleukin-10(IL-10)and interleukin-12(IL-12)in rat serum.The expressions of GATA3 and T-bet were detected by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).The expressions of GATA3 and T-bet in lung tissue were detected by immunohistochemistry(IHC).Result:Compared with the control group,the serum levels of IL-10 in the model group was significantly decreased,while the IL-12 level was significantly increased(P<0.01).The protein and gene expressions of GATA3 in lung tissue were significantly decreased,whereas the expression of T-bet was significantly increased(P<0.01).Compared with the model group,the serum level of IL-10 in each treatment group was increased to varying degrees,while the level of IL-12 was decreased to varying degrees(P<0.05).The mRNA and protein expressions of GATA3 in lung tissue of rats in each treatment group were increased significantly,while T-bet was inhibited(P<0.05).Conclusion:Modified Erchentang may reduce the inflammation of lung tissue and improve lung function in COPD rats by reducing IL-12,increasing the content of IL-10,inhibiting the protein and gene expressions of T-bet,and stimulating the protein and gene expressions of GATA3.