含羞草碱调控Notch1/Twist1信号通路改善博莱霉素诱导的大鼠肺纤维化  

L-mimosine ameliorates bleomycin-induced pulmonary fibrosis in rats through inhibiting Notch1/Twist1 signaling pathway

作　　者：何晓伟 汤娟 陈晓 李先伟 He Xiaowei;Tang Juan;Chen Xiao;Li Xianwei(Department of pharmacology,school of pharmacology,Wannan Medical College,Wuhu,Anhui,241002,China.)

机构地区：[1]皖南医学院药学院药理学教研室,安徽芜湖241002

出　　处：《齐齐哈尔医学院学报》2022年第1期6-12,共7页Journal of Qiqihar Medical University

基　　金：安徽省卫生健康委科研重点项目(2021zy033);安徽省高校自然科学研究重点项目(KJ2018A0254)。

摘　　要：目的研究含羞草碱(L-Mim)是否通过影响Notch1/Twist1信号通路,进而抑制肺泡上皮细胞间质转分化(EMT)而改善大鼠肺纤维化。方法雄性SD大鼠60只,随机分为正常组(CON组)、博莱霉素(BLM)组、L-Mim5 mg· kg^(-1)剂量组和L-Mim10 mg· kg^(-1)剂量组,每组各15只。CON组不做任何处理;其余三组气管注射BLM(7500 U· kg^(-1))诱导肺纤维化大鼠模型。造模后第3天开始灌胃给药,连续给药28天后检测用力肺活量(FVC)、第0.3秒用力呼气容积(FEV)占用力肺活量百分比(FEV0.3/FVC%)、最大呼气流量(PEF)和最大通气量(MVV)。取左肺行HE、Masson及I型胶原(collagen I)免疫组化染色;右肺提取RNA和蛋白,real-time PCR和(或)Western Blot检测肺组织中TGF-β1、E钙黏蛋白(E-Cadherin)、紧密连接蛋白-1(ZO-1)、N-钙黏蛋白(N-Cadherin)、波形蛋白(Vimentin)、α-平滑肌肌动蛋白(α-SMA)、Notch1、Notch1胞内域(NICD)、Hes1、Twist1、真核翻译起始因子3a(eIF3a)mRNA和蛋白的表达。结果与CON组相比,呼吸功能实验显示BLM组大鼠FVC、FEV0.3/FVC%、PEF和MVV明显降低。肺组织肺泡萎缩、塌陷并发生融合,肺泡间隔明显增宽,可见大量炎性细胞的浸润。肺间质胶原纤维沉积和collagen I的阳性表达明显增多;另外,肺组织中E-cadherin和ZO-1表达明显下降而Vimentin、N-cadherin和α-SMA的表达明显增加(P<0.01);同时,肺组织TGF-β1、Notch1、NICD、Hes1、Twist1和eIF3a的表达也明显上调(P<0.01)。与BLM组相比,不同剂量L-Mim能够明显改善大鼠的呼吸功能(FVC、FEV0.3/FVC%、PEF和MVV明显升高),显著降低肺组织炎症反应、胶原沉积和collagen I的阳性表达;另外,L-Mim还能够明显上调肺组织E-cadherin和ZO-1的表达和显著下调Vimentin、N-cadherin和α-SMA的表达;同时,L-Mim也能够明显抑制肺组织TGF-β1、Notch1、NICD、Hes1、Twist1和eIF3a的表达(P<0.05、0.01)。结论L-Mim能够逆转肺泡上皮细胞EMT、缓解肺纤维化。其机制可能与其抑�Objective To investigate whether the amelioration of L-mimosine(L-Mim)against pulmonary fibrosis(PF)by inhibiting epithelial-mesenchymal transition(EMT)of alveolar epithelial cells through affecting the Notch1/Twist1 signaling pathway.Methods Sixty male SD rats were randomly divided into control(CON)group,bleomycin(BLM)group,L-Mim 5 mgkg^(-1) group and L-Mim 10 mg·kg^(-1) group,15 rats in each group.The PF rat model was induced by intratracheal injection of BLM(7500 Ukg^(-1))in BLM group and BLM+L-Mim(5,10 mgkg^(-1))groups except CON group.L-Mimosine was administered on day 3 after BLM treatment,the forced vital capacity(FVC),the forced expiratory volume(FEV)in 0.3 s/forced vital capacity(FEV0.3/FVC%),peak expiratory flow(PEF)and maximal voluntary ventilation(MVV)of rats was evaluated for evaluating the respiratory function after 28 day.Excised inferior lobe of left lung was fixed in 10%formalin for HE staining,Masson staining and collagen I immunohistochemistry staining.The mRNA and(or)proteins levels of TGF-β1,E-Cadherin,ZO-1,Vimentin,E-Cadherin,α-SMA,Notch1,NICD,Hes1,Twist1 and eIF3a were detected by qPCR and(or)Western blotting in right lung tissue.Results The respiratory function tests showed that the FVC,FEV0.3/FVC%,PEF and MVV of rats were significantly lower in the BLM groups compared with the control group.The alveolar atrophy,collapse and fusion occurred,alveolar septum widened significantly,and a large number of inflammatory cells infiltrated in the pulmonary interstitial of the rats in the BLM group.And compared with control group,BLM obviously increased in collagen deposition and collagen I positive expression;the expression of ZO-1 and E-cadherin in lung tissues were decreased,and the expression of Vimentin,N-cadherin andα-SMA were increased(P<0.01);and concomitantly with increasing TGF-β1,Notch1,NICD,Hes1,Twist1 and eIF3a expression in lung tissues of rats.Compared with BLM group,the FVC,FEV0.3/FVC%,PEF and MVV of rats were significantly higher.L-Mim treatment significantly reduced the degree

关 键 词：含羞草碱 肺纤维化 上皮间质转分化 NOTCH1 TWIST1 

分 类 号：R966[医药卫生—药理学]