碱性成纤维细胞生长因子对干眼小鼠模型眼表白细胞介素-10和白细胞介素-1β蛋白表达的影响

Effects of basic fibroblast growth factor on the expression of interleukin-10 and interleukin-1β in ocular surface of dry eye mice model

目的研究碱性成纤维细胞生长因子(b FGF)对干眼小鼠模型眼表白细胞介素-10(IL-10)和白细胞介素-1β(IL-1β)蛋白表达的影响。方法将30只BALB/c小鼠构建干眼小鼠模型,并将建模成功的25只小鼠分为模型组(n=8,16眼)、阳性对照组(n=8,16眼)和实验组(n=9,18眼),另选10只未给予任何处理的BALB/c小鼠作为正常组。实验组给予b FGF滴眼,每次5μL,每天3次,阳性对照组给予普拉洛芬滴眼,每次25μL,每天3次,模型组和正常组给予无菌生理盐水滴眼,连续干预5 d。观察小鼠干眼症状指标水平,用苏木精-伊红(HE)染色法观察小鼠眼球角膜组织病理变化;用过碘酸-希夫(PAS)染色法观察小鼠结膜杯状细胞数量;用酶联免疫吸附(ELISA)法检测小鼠眼表组织中IL-10和IL-1β蛋白表达。结果干预后,实验组角膜荧光素钠染色评分、角膜炎症指数、泪膜破裂时间(BUT)和泪液分泌实验(SIt)分别为(10.22±2.06)分,0.09±0.02,(4.52±0.26)s,(5.26±0.52)mm/5 min,模型组分别为(11.02±2.25)分,0.52±0.06,(2.02±0.21)s,(4.00±0.32)mm/5 min,差异均有统计学意义(均P<0.01)。实验组和阳性对照组角膜上皮细胞完整、光滑,结膜穹窿部杯状细胞数量较模型组明显增多。实验组小鼠眼表组织中IL-10和IL-1β蛋白含量分别为(573.15±22.13),(174.25±12.36)μg·g^(-1),模型组分别为(236.48±41.28),(223.51±23.48)μg·g^(-1),差异均有统计学意义(均P<0.01)。结论 b FGF对小鼠干眼模型具有一定的治疗作用,可缓解干眼小鼠临床症状,抑制眼表鳞状上皮化生,其机制可能与其能够调节干眼眼表炎症反应有关。

Objective To investigate the effect of basic fibroblast growth factor（ b FGF） on the expression of interleukin-10（ IL-10） and interleukin-1β（ IL-1β） in ocular surface of dry eye mice model.Methods Thirty BALB/c mice were used to construct dry eye model,and 25 successful models were divided into model group（ n = 8,16 eyes）,positive control group（ n = 8,16 eyes） and test group（ n = 9,18 eyes）,10 BALB/c mice did not give any treatment as normal group.Test group was given b FGF 5 μL,3 times a day,and the positive control group was treated with piloprofen eye drops,25 μL a time,3 times·d^-1,the model group and normal group were given aseptic saline,and all groups were treated for 5 d. To observe the levels of dry eye symptomsindex in mice,hematoxylin-eosin（ HE） staining was used to observe the pathological changes of the cornea in the eye,the number of conjunctival goblet cells was observed by periodic acid-Schiff（ PAS） staining,the expression of IL-10 and IL-1β in ocular surface of mice were detected by enzyme-linked immunosorbent assay（ ELISA） method.Results After intervention,the corneal fluorescein sodium staining,corneal inflammation index,tear break-up time（ BUT） and tear Schirmer Ⅰ test（ SIt） in test group were 10. 22 ± 2. 06,0. 09 ± 0. 02,（ 4. 52 ± 0. 26） s,（ 5. 26 ± 0. 52） mm/5 min, had significant difference with those in model group, which were 11. 02 ± 2. 25,0. 52 ± 0. 06,（ 2. 02 ± 0. 21） s,（ 4. 00 ± 0. 32） mm/5 min（ all P〈0. 01）. The corneal epithelial cells in test group and positive control group were intact,smooth,the number of goblet cells in conjunctival fornix of test group and positive control group were significantly higher than that in model group. The levels of IL-10 and IL-1β protein in ocular surface of test group were（ 573. 15 ± 22. 13）,（ 174. 25 ± 12. 36） μg ·g^-1,had significant difference with those in model group,which were（ 236. 48 ± 41. 28）,（ 223. 51 ± 23. 48） μg·g^-1（ all P〈0. 01）. Conclusion b FGF can alleviate the clinical symptoms of dry eye in mice and inhibit the squamous metaplasia of the ocular surface. The mechanism may be related to its ability to regulate the inflammation of the dry eye.