固本防哮饮对哮喘缓解期小鼠肺组织线粒体凋亡相关因子CytC、Bcl-2及Caspase-3表达的影响

Effects of Guben Fangxiao Decoction on Mitochondrial Apoptosis-related Factors CytC,Bcl-2 and Caspase-3 in Lung Tissue of Mice in Remission of Asthma

目的观察固本防哮饮对哮喘缓解期小鼠肺组织线粒体凋亡相关因子细胞色素C(CytC)、B细胞淋巴瘤-2(Bcl-2)及半胱氨酸蛋白酶-3(Caspase-3)表达的影响,探讨其治疗哮喘的作用机制。方法将36只Balb/c小鼠随机分为正常组、模型组、孟鲁司特钠组及固本防哮饮低、中、高剂量组,每组6只。采用卵蛋白+氢氧化铝腹腔注射及卵蛋白雾化吸入法制备哮喘缓解期小鼠模型,造模成功后各给药组分别予相应药液灌胃,正常组和模型组灌胃等体积蒸馏水,连续30 d。Masson染色观察小鼠肺组织气道内胶原沉积情况,qPCR、免疫组化染色及Western blot检测肺组织CytC、Bcl-2、Caspase-3 mRNA和蛋白表达,ELISA检测肺组织白细胞介素(IL)-17A、神经生长因子(NGF)含量。结果与正常组比较,模型组小鼠肺组织炎性细胞浸润及气道内胶原纤维沉积明显增加,肺组织CytC mRNA和蛋白表达及Caspase-3蛋白表达显著升高(P<0.05,P<0.01),Bcl-2 mRNA和蛋白表达显著降低(P<0.05,P<0.01),IL-17A、NGF含量显著增加(P<0.05,P<0.01);与模型组比较,固本防哮饮各剂量组和孟鲁司特钠组小鼠肺组织气道内胶原纤维沉积有所改善,CytC mRNA表达显著降低(P<0.01),固本防哮饮低、中剂量组肺组织CytC、Caspase-3蛋白表达显著降低(P<0.05,P<0.01),Bcl-2 mRNA和蛋白表达显著升高(P<0.05,P<0.01),固本防哮饮高剂量组Caspase-3mRNA表达显著降低(P<0.05),固本防哮饮各剂量组NGF含量显著减少(P<0.05,P<0.01),固本防哮饮低剂量组IL-17A含量显著减少(P<0.05)。结论固本防哮饮可能通过调控线粒体凋亡通路抑制气道上皮细胞凋亡,减轻气道重塑,防治哮喘。

Objective To investigate the effects of Guben Fangxiao Decoction on CytC,Bcl-2 and Caspase-3 in lung tissue of mice in asthma remission stage;To discuss its mechanism in the treatment of asthma.Methods Totally 36 Balb/c mice were randomly divided into normal group,model group,montelukast sodium group and Guben Fangxiao Decoction low-,medium-and high-dosage groups,with 6 mice in each group.A mouse model of asthma in remission stage was established by intraperitoneal injection of ovalbumin+aluminum hydroxide and aerosol inhalation of ovalbumin.After the establishment of the model,each administration group was given corresponding drug solution by gavage,while the normal group and model group were given equal volume of distilled water by gavage for 30 consecutive days.Masson staining was used to observe the collagen deposition of airway in lung tissue,the mRNA and protein expressions of CytC,Bcl-2,Caspase-3 in lung tissue were detected by qPCR,immunohistochemistry and Western blot,the contents of IL-17A and NGF in lung tissue were detected by ELISA.Results Compared with the normal group,the inflammatory cell infiltration and collagen fiber deposition in lung tissue of the model group significantly increased,the expressions of CytC mRNA and protein and Caspase-3 protein in lung tissue significantly increased(P<0.05,P<0.01),the expressions of Bcl-2 mRNA and protein significantly decreased(P<0.05,P<0.01),and the contents of IL-17A and NGF significantly increased(P<0.05,P<0.01).Compared with the model group,the deposition of collagen fibers in lung tissue in each dosage of Guben Fangxiao Decoction group and montelukast sodium group were improved,the expressions of CytC mRNA significantly decreased(P<0.01),and Caspase-3 protein in lung tissue in Guben Fangxiao Decoction low-and medium-dosage groups significantly decreased(P<0.05,P<0.01),the expressions of Bcl-2 mRNA and protein significantly increased(P<0.05,P<0.01),the expression of Caspase-3 mRNA in Guben Fangxiao Decoction high-dosage group significantly decreased(P<0.05),the content of NGF in Guben Fangxiao Decoction each dosage groups significantly decreased(P<0.05,P<0.01),the content of IL-17A in Guben Fangxiao Decoction low-dosage group significantly decreased(P<0.05).Conclusion Guben Fangxiao Decoction may regulate mitochondrial apoptosis pathway,inhibit apoptosis of airway epithelial cells,and alleviate airway remodeling,in order to prevent and treat asthma.