复方苦参注射液对放射性肺损伤的肺保护作用

Lung Protective Mechanism of Compound Kushen Injection on Radiation-induced Pulmonary Injury

目的:观察复方苦参注射液对放射性肺损伤模型模型小鼠转化生长因子-β1(TGF-β1),Smad3蛋白(Smad3),糖原合成酶激酶-3β(GSK-3β),β-连环蛋白(β-catenin)表达的影响,并探讨其作用机制。方法:使用美国XStrahl小动物精准放疗辐照研究平台(SARRP),实施单次20 Gy双侧肺野照射建立放射性肺损伤小鼠模型,将32只小鼠随机分为正常组、模型组、复方苦参注射液组(5 g·kg^-1),地塞米松注射液组(0.5 mg·kg^-1)。正常组和模型组给予等体积0.9%氯化钠溶液,连续腹腔注射4周。苏木素-伊红(HE)染色观察肺组织病理;免疫组化(IHC)检测小鼠肺组织E-钙粘蛋白(E-cadheren),波形蛋白(Vimentin)表达;实时荧光定量聚合酶链式反应(Real-time PCR)检测组小鼠肺组织中TGF-β1,Smad3,GSK-3β和β-catenin mRNA的表达;蛋白免疫印迹法(Western blot)检测组小鼠肺组织中TGF-β1,Smad3,GSK-3β和β-catenin通道蛋白的表达。结果:与正常组比较,模型组小鼠肺系数显著升高(P<0.01),且小鼠肺脏出现炎症细胞浸润,肺间质水肿、充血,肺泡结构破坏,部分肺泡萎缩。与模型组比较,复方苦参注射液组小鼠肺脏炎症细胞浸润及肺间质病变减轻,小鼠肺组织E-cadheren的水平显著升高(P<0.01),小鼠肺组织Vimentin的水平显著降低(P<0.01),TGF-β1,Smad3,GSK-3β和β-catenin表达水平显著降低(P<0.01)。与地塞米松注射液组比较,复方苦参注射液小鼠肺部病理形态学改变相仿,E-cadheren,Vimentin,TGF-β1,Smad3,GSK-3β和β-catenin表达水平未见明显差异。结论:复方苦参注射液能够改善放射性肺损伤模型小鼠肺纤维化,其机制可能与抑制上皮间质转化(EMT)相关的TGF-β1,Smad3,GSK-3β和β-catenin mRNA及通道蛋白表达,进而促进E-cadheren表达、抑制Vimentin蛋白表达,最终抑制EMT的发生有关。

Objective:To observe the effect of compound Kushen injection on the expressions o transforming growth factor-β1(TGF-β1),drosophila mothers against decapentaplegic protein 3(Smad3),glycogen synthase kinase-3β(GSK-3β)andβ-catenin mice models with radiation-induced pulmonary injury(RIPI),in order to explore its possible mechanism of action.Method:On XStrahl precision radiation research platform for small animals(SARRP),a single 20 Gy bilateral lung field irradiation was performed to establish a mice model of RIPI.Thirty-two mice were randomly divided into normal control group,model group,compound Kushen injection group and dexamethasone injection group.The normal control group and the model group were given an equal volume of 0.9%sodium chloride solution and injected intraperitoneally for 4 weeks.The pathology of lung tissue tissues was observed by using hematoxylin-eosin(HE)staining.Immunohistochemical(IHC)was used to detect the expressions of E-cadheren and Vimentin proteins in mice lung tissues.Real-time polymerase chain reaction(Real-time PCR)was used to detect the mRNA expressions of TGF-β1,Smad3,GSK-3βandβ-cateninin.Western blot was used to detect the protein expressions of TGF-β1,Smad3,GSK-3βandβ-cateninin.Result:Compared with the normal group,the pulmonary coefficient of the model group was significantly decreased(P<0.01).Inflammatory cell infiltration,pulmonary interstitial edema,congestion,destruction of alveolar structure and partial alveolar atrophy were observed in the lung tissues of the model group.Compared with the model group,in the compound Kushen injection group,the levels of infiltration of lung inflammatory cells and pulmonary interstitial lesions in mice,the expression of Vimentin in lung tissues(P<0.01),and the expressions of TGF-β1,Smad3,GSK-3βandβ-cateninin were significantly decreased(P<0.01),whereas the expression of E-cadheren was significantly increased(P<0.01).However,compared with the dexamethasone injection group,in the compound Kushen injection group,the pathological changes of lung tissues were similar,and the expression levels of E-cadheren,Vimentin,TGF-β1,Smad3,GSK-3βandβ-cateninin were not significantly different.Conclusion:Compound Kushen injection can alleviate pulmonary fibrosis of lung in the treatment of RIPI,and the mechanism may be associated with inhibiting the mRNA and protein expressions of TGF-β1,Smad3,GSK-3βandβ-catenin related to epithelial-mesenchymal transition(EMT),promoting the expression of E-cadheren,and inhibiting the expression of Vimentin,so as to inhibit the occurrence of EMT.