针刺对去势雌兔干眼症模型眼表粘蛋白分泌表达的影响及其机制研究

Impact of Acupuncture on Expression of Secretion in Castrated Female Rabbits for Dry Eye Model and Its Mechanism

目的:研究针刺治疗手段对去势雌兔干眼症模型中的眼表粘蛋白分泌表达的影响及其相关机制。方法:利用动物试验,通过分组对照研究,观察针刺前后更年期干眼症模型去势雌兔及正常兔眼表损伤程度的变化。结合结膜印迹细胞学检查技术,采用免疫组化SABC法检测眼表结膜组织MUC5AC、MUC19蛋白表达的差异,对针刺前后干眼症模型兔及正常兔眼表蛋白质成分的改变进行定量分析,探讨针刺对兔眼表蛋白质的影响以及可能的靶器官及相关作用机制。SPSS20.0软件对实验结果进行统计分析。结果:去势模型的建立成功,在去势建模的基础上,实验组和空白组在Shirmer试验中P<0.05,有显著差异,角膜荧光染色法和虎红染色检查也表明去势后实验组与空白对照组有差异;HE染色发现:针刺前腺小叶清晰,腺体胞浆丰富,少见空泡;针刺后结构依旧保持完整且腺上皮结构清晰,但腺小叶中腺泡内物质大量排空,提示分泌旺盛;针刺前后兔泪腺实行了电镜观察:针刺后泪腺中的高尔基体丰富;针刺后的泪腺组织VIP浓度:109.75±22.32pg/100g组织,针刺前VIP浓度为62.71±20.12=pg/100g,有显著性统计学差异(P<0.0001);去势前后MUC5AC、MUC19蛋白表达有显著性差异,且随着针刺治疗的进展,杯状细胞计数逐渐增加,结膜细胞鳞状上皮化生减少,MUC5AC、MUC19蛋白表达增加,靠近于空白组。结论:在接受针刺治疗干眼症患者中,结膜细胞鳞状上皮化生的情况有所改善,杯状细胞数量增加,且可以提高MUC5AC和MUC19蛋白的表达。

Objective：To investigate the impact on the expression of secretion in castrated female rabbits and its mechanism during acupuncturing dry eye model.Methods：Through grouping controlled studies,using animal trials to observe dry eye model ovariectomized rabbits＇ and normal rabbits＇ ocular surface damage ocular before and after acupuncturing,combined with conjunctival impression cytology technique,using immunohistochemical SABC method to detect ocular conjunctiva MUC5 AC and MUC19 protein expression to see the differences in the rabbits before and after acupuncturing,in order to analyze the effect of protein changes before and after acupuncturing on ocular surface,finding possible target organs and related mechanisms.Experimental data were statistically analyzed using SPSS20.0 software.Results：Successfully established the ovariectomized model.On the basis of castration modeling,in the Shirmer trial,the experimental group and the controlled group,showed P〈0.05,there were significant differences,corneal fluorescent staining and HE staining also showed differences,between the experimental group and the blank controlled group;HE staining showed that before acupuncturing,structure of glandular lobules was clear,abundant gland cytoplasm,vacuoles were rare;After acupuncturing,complete needled structure and clear glandular structure were observed,acinar gland lobules drained a lot of substance,secreted exuberant;After acupuncturing,rabbit lacrimal gland introduced electron microscopy had found that lacrimal gland Golgi were richer than before;Lacrimal gland tissue vasoactive intestinal peptide concentration [（109.75±22.32） pg/100 g tissue] was significantly higher than that before acupuncturing [（62.71±20.12） pg/100 g organization],there were statistically significant difference（P〈0.0001）;MUC5 AC and MUC19 protein expression were significantly different between the experimental and the blank controlled groups,and with the progress of acupuncture,the number of goblet cell count increased,conjunctival squamous cell metaplasia had reduced,MUC5 AC and MUC19 expression were increasing,much closer to the controlled group.Conclusion：Acupuncture treatment helps to reduce conjunctival squamous cell metaplasia,increase the number of goblet cells and increases MUC5 AC,MUC19 expression.