M细胞对过敏性结膜炎小鼠MEK/ERK信号通路的影响

Effect of M cells on MEK/ERK signaling pathway in mice with allergic conjunctivitis

目的探索M细胞在过敏性结膜炎小鼠中与丝裂原活化蛋白/细胞外调节蛋白激酶(MEK/ERK)信号通路的联系。方法Barb/c小鼠随机分为空白对照组、模型组和治疗组,每组12只。空白对照组正常饲养,模型组和治疗组使用卵清蛋白(OVA)建立小鼠过敏性结膜炎模型。治疗组采用盐酸奥洛他定滴眼液治疗,每天滴加2次,每次1滴;模型组滴加等量生理盐水,空白对照组不做处理。苏木精-伊红(HE)染色检测小鼠眼睑组织病理变化情况,实时荧光聚合酶链式反应(RT-PCR)法检测眼睑中M细胞标记物糖蛋白2(GP2)、趋化因子配体9(CCL9)和细胞外调节蛋白激酶(ERK)mRNA相对表达量,免疫印迹试验(WB)法检测眼睑中GP2、CCL9、ERK蛋白相对表达量。结果HE染色显示,与模型组比较,治疗组眼睑组织较为紧密,无炎症细胞浸润,无明显水肿,固有层嗜酸性粒细胞及中性粒细胞浸润较弱,肥大细胞数量明显降低,脱颗粒现象不明显。RT-PCR结果表明,与空白对照组比较,模型组小鼠眼睑中GP2、CCL9、ERK mRNA相对表达量显著上升,差异有统计学意义(P<0.05);与模型组比较,治疗组小鼠眼睑中GP2、CCL9、ERK mRNA相对表达量显著下降,差异有统计学意义(P<0.05);空白对照组与治疗组小鼠眼睑中GP2、CCL9、ERK相对表达量差异不明显(P>0.05)。Western blot结果表明,与空白对照组比较,模型组小鼠眼睑中GP2、CCL9、ERK蛋白相对表达量显著上升,差异有统计学意义(P<0.05);与模型组比较,治疗组小鼠眼睑中GP2、CCL9、ERK蛋白相对表达量显著下降,差异有统计学意义(P<0.05);空白对照组与治疗组小鼠眼睑中GP2、CCL9、ERK蛋白相对表达量差异不明显(P>0.05)。结论M细胞增殖可能与MEK/ERK信号通路的激活相关。

Objective To explore the relationship between M cells and the mitogen-activated protein/extracellular regulatory protein kinase(MEK/ERK)signaling pathway in mice with allergic conjunctivitis.Methods Barb/c mice were randomly divided into blank control group,model group,treatment group,and 12 mice in each group.The blank control group was feed normally.Ovalbumin(OVA)was used to establish a model of allergic conjunctivitis in mice in model group and treatment group.The treatment group was treated with olopatadine hydrochloride eye drops,which was dripped twice a day,1 drop each time;the model group was dripped with the same amount of normal saline,and the blank control group was left untreated.Hematoxylin-Eosin(HE)staining was used to detect the pathological changes of mouse eyelid tissues.Real-time fluorescent polymerase chain reaction(RT-PCR)method was used to detect the relative expression of M cells markers glycoprotein 2(GP2),chemokine(C-C motif)ligand 9(CCL9),and extracellular regulatory protein kinase(ERK)mRNA in the eyelids.Western blot test(WB)was used to detect the relative expression of GP2,CCL9,and ERK protein in the eyelids.Results HE staining showed that compared with the model group,the eyelid tissue of the treatment group was tighter,without inflammatory cell infiltration,no obvious edema,weaker infiltration of eosinophils and neutrophils in the lamina propria,significantly reduced number of mast cells,and no obvious degranulation.RT-PCR results showed that compared with the blank control group,the relative expression of GP2,CCL9 and ERK mRNA in the eyelids of the model group increased significantly(P<0.05);compared with the model group,the relative expression of GP2,CCL9 and ERK mRNA in the eyelids of the treatment group decreased significantly(P<0.05).The relative expressions of GP2,CCL9 and ERK in the eyelids of the blank group and the treatment group were not significantly different(P>0.05).Western blot results showed that compared with the blank control group,the relative expression of GP2,CCL9 and ERK protein in the eyelids of the model group increased significantly(P<0.05);compared with the model group,the relative expression of GP2,CCL9 and ERK protein in the eyelids of the treatment group decreased significantly(P<0.05).The relative expression of GP2,CCL9 and ERK protein in the eyelids of the blank group and the treatment group were not significantly different(P>0.05).Conclusion M cell proliferation was associated with the pathogenesis of allergic conjunctivitis,and its proliferation might be associated with activation of the MEK/ERK signaling pathway.