阳和平喘颗粒对哮喘BMSCs归巢及Notch信号通路的影响

Effect of Yanghe Pingchuan Granule on Homing and Notch Signaling Pathway of Asthma BMSCs

目的观察阳和平喘颗粒通过调控Notch信号通路对骨髓间充质干细胞(BMSCs)移植后哮喘大鼠气道炎症的影响。方法取30 SD只大鼠随机分为5组:正常对照(NC)组、模型对照(MC)组、BMSCs移植(BMSCs)组、BMSCs地塞米松组(BMSCs地米组)、BMSCs阳和平喘颗粒组(BMSCs阳和组),每组6只。给予大鼠卵清蛋白致敏激发哮喘模型,尾静脉移植BMSCs。计数支气管肺泡灌洗液(BALF)中嗜酸性粒细胞(E)、嗜中性粒细胞(Neu)、淋巴细胞(L)、单核细胞(MΦ),酶联免疫吸附法检测肺组织白细胞介素(IL)-2、IL-4、CXC趋化因子受体(CXCR-3)和CC趋化因子受体4(CCR-4),RT-PCR法检测肺组织Notch通路配体Jagged1及相关基因重组信号序列结合蛋白-J(RBP-J)、GATA结合蛋白3(GATA-3)mRNA表达,免疫印迹法检测肺组织基质细胞衍生因子-1(SDF-1)、CXC趋化因子受体(CXCR)-4蛋白表达。结果与NC组比校,MC组BALF中炎性细胞总数及各炎性细胞计数显著升高,IL-2、CXCR-3、SDF-1表达降低,IL-4、CCR-4表达升高,肺组织Notch1、Jagged1、RBP-J、GTAT-3表达升高(P<0.05,P<0.01)。与MC组比较,BMSCs地米组及BMSCs阳和组炎性细胞(E、Neu、L、MΦ)数目均减少,IL-2、CXCR-3、SDF-1、CXCR4表达升高,IL-4、CCR-4降低,肺组织Notch 1、Jagged 1、RBP-J、GTAT-3表达降低(P<0.05,P<0.01)。与BMSCs组比较,BMSCs地米组MΦ数目、IL-4、CCR-4、Notch1、RBP-J表达降低,BMSCs阳和组Neu、MΦ、IL-4、Notch1、RBP-J表达降低(P<0.05)。BMSCs地米组与BMSCs阳和组比较,差异无统计学意义(P>0.05)。结论阳和平喘颗粒可通过调节Notch通路,促进BMSCs向哮喘肺组织的归巢率,强化对Th2炎症的抑制作用而改善哮喘。

Objective To observe the effect of Yanghe Pingchuan Granule(YHPCG)on the airway inflammation of asthmatic rats after transplantation of bone mesenchymal stem cells(BMSCs)by regulating Notch signaling pathway.Methods Thirty SD rats were randomly divided into 5 groups:normal control(NC)group,model control(MC)group,BMSCs transplantation(BMSCs)group,BMSCs dexamethasone group,BMSCs YHPCG group,6 in each group.The asthma model was sensitized with ovalbumin and then BMSCs were transplanted into the rats via the tail vein.The numbers of eosinophils(E),neutrophils(Neu),lymphocytes(L),and monocytes(MΦ)in bronchoalveolar lavage fluid(BALF)were counted.The levels of IL-2,IL-4,CXC chemokine receptor-3(CXCR-3)and CC chemokine receptor-4(CCR-4)in lung tissue were detected by ELISA.RT-PCR was used to detect the expressions of Notch pathway ligand Jagged1 and relevant gene recombination signal sequence binding protein-J(RBP-J),GATA binding protein 3(GATA-3)mRNA.Western blot was used to detect the expressions of stromal cell-derived factor-1(SDF-1)and CXC chemokine receptor 4(CXCR4)protein in lung tissue.Results Compared with the NC group,the total number of inflammatory cells and the count of inflammatory cells in BALF increased significantly,as well as the expressions of IL-4,CCR-4,Notch1,Jagged1,RBP-J,GTAT-3 in lung tissue in the MC group(P<0.05,P<0.01).Compared with the MC group,the numbers of inflammatory cells(E,Neu,L,MΦ)the expressions of IL-4 and CCR-4,as well as the expressions of Notch1,Jagged1,RBP-J and GTAT-3 in lung tissue were lower in BMSCs dexamethasone group and BMSCs Yanghe group.Compared with the MC group,expressions of IL-2,CXCR-3,SDF-1,and CXCR4 were higher in BMSCs dexamethasone group and BMSCs Yanghe group(P<0.05,P<0.01).Compared with the BMSCs group,the numbers of MΦ,IL-4,CCR-4,Notch1,RBP-J in the BMSCs dexamethasone group decreased,and the expressions of Neu,MΦ,IL-4,Notch1 and RBP-J in the BMSCs Yanghe group decreased(P<0.05).There was no significant difference between BMSCs dexamethasone group and BMSCs Yanghe group(P>0.05).Conclusion YHPCG increased the homing rate of BMSCs to asthmatic lung tissue by regulating the Notch pathway,thereby strengthening the inhibitory effect of BMSCs on Th2 inflammation to improve asthma.