盐酸奥洛他定滴眼液对过敏性结膜炎小鼠结膜M细胞增殖及白细胞介素4、JAK1、STAT6表达的影响

Effects of olopatadine hydrochloride eye drops on proliferation of conjunctival M cells and interleukin 4,JAK1 and STAT6 expression in mice with allergic conjunctivitis

目的探讨盐酸奥洛他定滴眼液对过敏性结膜炎小鼠结膜M细胞增殖及白细胞介素4(interleukin 4,IL-4)、JAK1、STAT6表达的影响。方法选取SPF级Barb/c小鼠18只,随机分为空白对照组、模型组、治疗组,每组6只;用卵清蛋白和Al(OH)3对模型组和治疗组小鼠造模。造模后治疗组采用盐酸奥洛他定滴眼液治疗,每天滴眼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。采用免疫荧光法检测各组小鼠眼睑结膜M细胞数;ELISA检测各组小鼠血清中IL-4、JAK1、STAT6含量;RT-PCR检测眼睑组织中JAK1、STAT6 mRNA相对表达量;Western blot法检测眼睑组织中JAK1、STAT6蛋白相对表达量。结果眼睑结膜组织免疫荧光检测结果显示,模型组标本中M细胞阳性率远高于空白对照组(P<0.05);治疗组M细胞阳性率有所下降,低于模型组(P<0.05),但仍高于空白对照组(P<0.05);推测过敏性结膜炎病程与M细胞阳性率升高有关。ELISA检测结果显示,模型组血清中IL-4、JAK1、STAT6含量远高于空白对照组(均为P<0.05);治疗组IL-4、JAK1、STAT6含量均较模型组显著下降(均为P<0.05);治疗组血清中IL-4、JAK1、STAT6含量均稍高于空白对照组(均为P<0.05);推测过敏性结膜炎发病过程中伴随血清中IL-4、JAK1、STAT6含量变化。眼睑组织中,模型组JAK1、STAT6 mRNA以及蛋白相对表达量均远高于空白对照组(均为P<0.05);治疗组JAK1、STAT6 mRNA以及蛋白相对表达量较模型组显著下降(均为P<0.05),与空白对照组相比,差异均无统计学意义(均为P>0.05)。结论M细胞增殖与过敏性结膜炎病程具有相关性,其增殖可能与IL-4-JAK1-STAT6信号通路的激活相关。

Objective To investigate the effects of olopatadine hydrochloride on proliferation of conjunctival M cells and interleukin 4(IL-4),JAK1 and STAT6 expression in mice with allergic conjunctivitis.Methods Totally 18 BALB/c mice,special pathogenfree(SPF),were randomly divided into blank control group,model group and treatment group,6 mice in each group.Egg albumin(OVA)and AI(OH)3 were used to establish a model of allergic conjunctivitis in model group and treatment group.The mice in treatment group were treated with olopatadine hydrochloride eye drops,twice a day,1 drop per time.The mice in model group were treated with normal saline in the same dosage.The mice in blank control group received no treatment.The amount of M cells was detected by immunofluorescence.The contents of IL-4 and STAT6 in serum were measured by ELISA.The relative expression of mRNAs for JAK1 and STAT6 in blepharal tissue was discovered by PCR.The relative expression of JAK1 and STAT6 protein in blepharal tissue was investigated by Western blot.Results In blepharal tissue,immunofluorescence assay showed that the positive rate of M cells in the model group was much higher than that in the blank control group(P<0.05).The positive rate of M cells in the treatment group was lower than that in the model group(P<0.05),but higher than that in the blank control group(P<0.05).We speculated that the progress of allergic conjunctivitis may be related to the increased positive rate of M cells.In serum,ELISA results showed that the levels of IL-4,JAK1 and STAT6 in the model group were much higher than those in the blank control group(all P<0.05).The levels of IL-4,JAK1 and STAT6 in treatment group were significantly lower than those in the model group,but slightly higher than those in the blank control group(all P<0.05).We speculated that the levels of IL-4,JAK1 and STAT6 in serum were associated with the pathogenesis of allergic conjunctivitis.In the blepharal tissue,the mRNA and proteins levels of JAK1,STAT6 in the model group were much higher than those in the blank control group(all P<0.05).The mRNA and proteins levels of JAK1,STAT6 in the treatment group were significantly lower than those in the model group(all P<0.05),but there was no significant difference in the treatment group and the blank control group(all P>0.05).Conclusion The proliferation of M cells is related with the pathogenesis of allergic conjunctivitis,and may be associated with the activation of the IL-4,JAK1 and STAT6 signal pathway.