慢病毒介导的Jagged1 RNA干扰通过抑制肥大细胞的迁移、浸润和脱颗粒减轻变应性鼻炎小鼠的炎症

Lentivirus-mediated Jagged1 RNA Interference Reduces Inflammation in Allergic Rhinitis Mice by Inhibiting Mast Cell Migration,Infiltration and Degranulation

目的探究慢病毒介导的Jagged1 RNA干扰通过抑制肥大细胞的迁移,浸润和脱颗粒对变应性鼻炎小鼠炎症的影响。方法32只BALB/c小鼠随机分为4组:对照组、AR组、sh-NC组和sh-Jagged1组,每组各8只。卵清蛋白(OVA)联合氢氧化铝制备变应性鼻炎小鼠。RT-PCR检测小鼠Jagged1 mRNA表达;Western blot法检测小鼠Jagged1蛋白表达;鼻部症状评分评估小鼠变应性鼻炎症状;ELISA检测小鼠组胺、类胰蛋白酶、前列腺素D2(Prostaglandin D2)和IgE含量;HE染色观察鼻黏膜病理学变化;电子显微镜观察小鼠鼻黏膜纤毛结构变化;甲苯胺蓝染色观察小鼠鼻黏膜肥大细胞浸润。结果与对照组比较,AR组和sh-NC组小鼠的Jagged1表达、症状评分升高和IgE表达升高(P<0.01)。与sh-NC组比较,sh-Jagged1组小鼠Jagged1表达、症状评分降低和IgE表达降低(P<0.01)。与对照组比较,AR组和sh-NC组小鼠的鼻中隔黏膜基膜破碎,黏膜下血管出现明显扩张、充血现象,炎性细胞浸润和肥大细胞数量增加(P<0.01),纤毛大面积减少、排列紊乱,组胺、类胰蛋白酶和PGD2水平明显升高(P<0.01)。sh-Jagged1可改善AR小鼠鼻黏膜和纤毛损伤,抑制炎性细胞浸润、肥大细胞数量、组胺、类胰蛋白酶和PGD2水平(P<0.01)。结论慢病毒介导的sh-Jagged1通过抑制肥大细胞的迁移,浸润和脱颗粒减轻变应性鼻炎小鼠炎症。

Objective To investigate the effect of lentivirus-mediated Jagged1 RNA interference on inflammation in mice with allergic rhinitis by inhibiting mast cell migration,infiltration and degranulation.Methods Thirty-two BALB/c mice were randomly divided into 4 groups:control group,AR group,sh-NC group and sh-Jagged1 group,8 in each group.Ovalbumin(OVA)combined with aluminum hydroxide was used to prepare allergic rhinitis mice.RT-PCR was used to detect Jagged1 mRNA expression of mouse;Western blot was used to detect Jagged1 protein expression of mouse;nasal symptom score was used to evaluate allergic rhinitis symptoms in mice;ELISA was used to detect histamine,tryptase,prostaglandin D2(prostaglandin D2)and IgE content of mouse;HE staining was used to observe the pathological changes of nasal mucosa;electron microscopy was used to observe the changes of mouse nasal mucosa cilia structure;toluidine blue staining was used to observe the infiltration of mast cells in mouse nasal mucosa.Results Compared with the control group,the expression of Jagged1,symptom score and IgE increased in AR group and sh-NC group(P<0.01).Compared with the sh-NC group,the expression of Jagged1,symptom score and IgE decreased in the sh-Jagged1 group(P<0.01).Compared with the control group,the nasal septal mucosal basement membrane of the AR group and sh-NC group was broken,the submucosal blood vessels showed obvious dilation and hyperemia,the infiltration of inflammatory cells and the number of mast cells increased(P<0.01),and the cilia area was large Decreased,disordered arrangement,histamine,tryptase,and PGD2 levels were significantly increased(P<0.01).sh-Jagged1 can improve the nasal mucosa and cilia damage in AR mice,inhibit inflammatory cell infiltration,mast cell number,histamine,tryptase,and PGD2 levels(P<0.01).Conclusion Lentivirus-mediated sh-Jagged1 reduces inflammation in mice with allergic rhinitis by inhibiting mast cell migration,infiltration and degranulation.