黄芪免煎颗粒对兔干眼模型泪液及角膜、结膜黏蛋白1表达的影响

Effect of Astragalus membranaceus Granules on Mucin 1 expression in tear,cornea,and conjunctiva of rabbit dry eye model

目的观察益卫固表药物黄芪免煎颗粒对兔干眼模型泪液及角膜、结膜黏蛋白1(MUC1)表达的影响。方法选取健康成年家兔30只,随机分为对照组(CG)、模型组(MG)、黄芪组(AM),每组10只。MG组和AM组建立兔黏蛋白缺乏型干眼模型,AM组以黄芪免煎颗粒口服溶液8.0 mL/kg灌胃28 d,CG组和MG组以等量0.9%氯化钠注射液灌胃。于造模后第0、28、56 d取家兔泪液,酶联免疫吸附法检测MUC1表达情况。第56 d取角膜、结膜行免疫荧光法检测MUC1表达,实时荧光定量PCR(RT-qPCR)检测MUC1基因表达。结果(1)泪液MUC1:与CG组比较,MG组兔第28、56 d泪液MUC1表达降低(t_(28 d)=26.739,t_(56 d)=15.268,均P=0.000);与MG组比较,AM组兔第56 d泪液MUC1表达升高(t=8.031,P=0.000),差异均有统计学意义。(2)角膜、结膜MUC1基因:与CG组比较,MG组MUC1基因表达降低(t=19.261,P=0.000);与MG组比较,AM组MUC1基因表达升高(t=6.008,P=0.000),差异均有统计学意义。(3)角膜、结膜MUC1蛋白:与MG组比较,AM组角膜、结膜MUC1蛋白表达均升高(t_(角膜)=4.582,t_(结膜)=6.354,均P=0.000),差异有统计学意义。结论黄芪免煎颗粒可通过提高干眼模型兔的泪液及角膜、结膜MUC1蛋白表达,发挥补益卫气作用,缓解黏蛋白缺乏型干眼症状。

OBJECTIVE To observe the expression of mucin 1(MUC1)in tear,cornea,and conjunctiva in rabbit dry eye model affected by Astragalus membranaceus Granules.METHODS Thirty healthy adult rabbits were randomly divided into control group(CG),model group(MG)and Astragalus membranaceus group(AM),with 10 rabbits in each group.The rabbit model of dry eye was established in MG and AM.After model establishment,rabbits in AM were fed with 8.0 mL/kg Astragalus membranaceus Granules oral solution by intragastric administration for 28 days,CG and MG were fed with the same volume of saline solution.The tear of rabbits were collected at the very day,the 28^(th) and 56^(th) day after modeling,then the expression of MUC1 was detected by enzyme-linked immunosorbent assay.The cornea and conjunctiva were collected at the 56th day after modeling,then the expression of MUC1 gene was detected by immunofluorescence and real-time fluorescence quantitative PCR(RT-qPCR).RESULTS(1)MUC1 in tear:Compared with CG,the expression of MUC1 in MG decreased on the 28^(th) and 56^(th) day(t_(28 d)=26.739,t_(56 d)=15.268,all P=0.000);Compared with MG,the expression of MUC1 in AM increased on the 56^(th) day(t=8.031,P=0.000).(2)MUC1 gene in corneas and conjunctiva:Compared with CG,the expression of MUC1 gene in MG decreased(t=19.261,P=0.000);Compared with MG,the expression of MUC1 gene in AM increased(t=6.008,P=0.000),and the differences were statistically significant.(3)MUC1 protein in cornea and conjunctiva:Compared with MG,the expression of MUC1 protein in cornea(t=4.582,P=0.000)and conjunctiva(t=6.354,P=0.000)in AM increased,and the differences were statistically significant.CONCLUSIONS Astragalus membranaceus Granules can improve the expression of MUC1 protein in tear,cornea,and conjunctiva of rabbit dry eye model by tonifying defense Qi and thereby alleviate symptoms of mucin deficiency dry eye.