普拉洛芬治疗苯扎氯铵诱导的小鼠干眼的实验研究
发布时间：2018-11-14 15:11
【摘要】：目的通过随机对照实验探讨普拉洛芬对苯扎氯铵诱导小鼠干眼的治疗作用及其可能机制。 方法选用BALB/c雄性小鼠70只,使用0.25%的苯扎氯铵溶液局部点眼以诱导干眼。在第21天时根据泪膜破裂时间、角膜荧光素钠染色、炎症指数等参数选择干眼表现程度相似的眼并随机分为A、B、C、D四组。A组为空白对照,B、C、D组分别使用0.1%玻璃酸钠滴眼液,0.1%玻璃酸钠滴眼液加0.1%氟米龙滴眼液,0.1%玻璃酸钠滴眼液加0.1%普拉洛芬滴眼液进行治疗。治疗后第0、1、3、5天进行泪膜破裂时间和角膜上皮荧光素钠染色,眼表炎症程度的评估及基础泪液量的测量。治疗结束后取下小鼠眼球,进行HE及过碘酸-希夫(periodic acid-schiff, PAS)染色,角蛋白10(cytokeratin 10, K10)免疫荧光标记,以及Western blot检测角结膜组织中肿瘤坏死因子α(tumor necrosis factor-α, TNF-α)表达水平。 结果共72眼入选治疗实验,每组18眼。在使用药物治疗后的第0、1、3天各项临床指标无统计学差异。治疗第5天时,A、B、C、D组泪膜破裂时间(BUT)分别为(2.933±0.320)、(2.900±0.280)、(3.464±0.498)和(3.643±0.413)秒;其中C组与D组的BUT均较A组和B组延长,差异具统计学意义(F=13.774,P=0.000)。角膜荧光素钠染色分级A、B、C、D组分别为(11.640±1.008)、(11.790±1.188)、(10.330±1.371)及(10.270±1.104)分;两联合用药治疗组均较人工泪液组和空白对照组角膜荧光素钠染色分级减轻,具有统计学意义(F=6.145,P=0.001)。角膜炎症指数评分A、B、C、D组分别为(0.232±0.059)、(0.229±0.078)、(0.151±0.055)及(0.154±0.056)分;两联合用药治疗组均较人工泪液组和空白对照组炎症指数降低,具有统计学意义(F=6.703,P=0.001)。各组间的基础泪液分泌量无显著性差异;C组与D组在泪膜破裂时间、角膜荧光素钠染色分级及眼表炎症程度中均无统计差异。HE结果显示空白对照组上皮厚度高于两联合用药组,联合用药组上皮规整;PAS染色示普拉洛芬组及氟米龙组的杯状细胞数量相当,均多于人工泪液组及空白对照组。普拉洛芬组及氟米龙组的角膜上皮层均几乎不表达K10,而人工泪液组及空白对照组仍阳性表达。普拉洛芬组及氟米龙处理组角膜中的TNF-α水平均有下降。 结论玻璃酸钠联合普拉洛芬或氟米龙对苯扎氯铵诱导的小鼠干眼具有相似效果,与单用玻璃酸钠相比,可更好地稳定泪膜,促进上皮修复,下调炎症因子TNF-α,增加杯状细胞数量,抑制鳞状上皮化生。普拉洛芬可以抑制苯扎氯铵诱导的小鼠干眼眼表炎症,有望用于临床中重度干眼的治疗。
[Abstract]:Objective to investigate the therapeutic effect of Praprofen on the dry eye of mice induced by benzalkonium chloride and its possible mechanism. Methods 70 BALB/c male mice were used to induce dry eye with 0.25% benzalkonium chloride solution. On the 21st day, according to tear film rupture time, corneal fluorescein sodium staining, inflammation index and other parameters, the dry eyes were randomly divided into four groups: group A (A) as blank control, group A (B) C, B (C), C (P < 0.05). Group D was treated with 0.1% sodium hyaluronate eye drops, 0.1% sodium hyaluronate eye drops plus 0.1% flumilon eye drops, 0.1% sodium hyaluronate eye drops and 0.1% Praprofen eye drops respectively. The time of lacrimal film rupture and corneal epithelium fluorescein sodium staining, the evaluation of ocular surface inflammation and the measurement of basic tear volume were performed 5 days after treatment. After treatment, the eyeballs of mice were removed and stained with HE and periodic acid-schiff, PAS, cytokeratin 10 (K10) immunofluorescent labeling and Western blot were used to detect tumor necrosis factor 伪 (tumor necrosis factor- 伪 in the cornea conjunctiva. TNF- 伪) expression level. Results A total of 72 eyes were enrolled in the treatment experiment, 18 eyes in each group. There was no statistical difference in the clinical indexes on the 1st day after drug therapy. On the 5th day of treatment, the time of tear film rupture in group D was (2.933 卤0.320), () 2.900 卤0.280), (3.464 卤0.498) seconds and (3.643 卤0.413) seconds, respectively. The BUT of group C and group D were significantly longer than that of group A and group B, and the difference was statistically significant. Corneal fluorescein sodium staining grade was (11.640 卤1.008), (), 11.790 卤1.188), (10.330 卤1.371) and (10.270 卤1.104), respectively. Compared with the artificial tear group and the blank control group, the grade of corneal fluorescein sodium staining in the combined treatment group was significantly lower than that in the artificial tear group and the blank control group (F _ (6.145) P ~ (0. 001). The scores of corneal inflammation index in group A were (0.232 卤0.059), (, 0.229 卤0.078), (, 0.151 卤0.055) and (0.154 卤0.056), respectively. The inflammatory index of the two combined groups was significantly lower than that of the artificial tear group and the blank control group (F = 6.703, P < 0.001). There was no significant difference in the amount of basal tear secretion among the groups. There was no statistical difference in tear film rupture time, corneal fluorescein sodium staining and ocular surface inflammation between group C and group D. HE results showed that the thickness of epithelium in the blank control group was higher than that in the combined treatment group, and the epithelium was regular in the combined treatment group. PAS staining showed that the number of goblet cells in Praprofen group and flumilon group was similar, which was more than that in artificial tear group and blank control group. K10 was almost not expressed in the corneal epithelium of the Praprofen group and flumilon group, but remained positive in the artificial tear group and the blank control group. The level of TNF- 伪 in cornea decreased in both Praprofen group and flumilon group. Conclusion Sodium hyaluronate combined with Praprofen or flumilon has similar effects on dry eye of mice induced by benzalkonium chloride. Compared with sodium hyaluronate alone, sodium hyaluronate can stabilize tear film, promote epithelial repair and down-regulate the inflammatory factor TNF- 伪. Increase the number of goblet cells and inhibit squamous metaplasia. Praprofen can inhibit the inflammation of dry eye surface induced by benzalkonium chloride in mice and may be used in the treatment of moderate and severe dry eye.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R777
本文编号：2331527
[Abstract]:Objective to investigate the therapeutic effect of Praprofen on the dry eye of mice induced by benzalkonium chloride and its possible mechanism. Methods 70 BALB/c male mice were used to induce dry eye with 0.25% benzalkonium chloride solution. On the 21st day, according to tear film rupture time, corneal fluorescein sodium staining, inflammation index and other parameters, the dry eyes were randomly divided into four groups: group A (A) as blank control, group A (B) C, B (C), C (P < 0.05). Group D was treated with 0.1% sodium hyaluronate eye drops, 0.1% sodium hyaluronate eye drops plus 0.1% flumilon eye drops, 0.1% sodium hyaluronate eye drops and 0.1% Praprofen eye drops respectively. The time of lacrimal film rupture and corneal epithelium fluorescein sodium staining, the evaluation of ocular surface inflammation and the measurement of basic tear volume were performed 5 days after treatment. After treatment, the eyeballs of mice were removed and stained with HE and periodic acid-schiff, PAS, cytokeratin 10 (K10) immunofluorescent labeling and Western blot were used to detect tumor necrosis factor 伪 (tumor necrosis factor- 伪 in the cornea conjunctiva. TNF- 伪) expression level. Results A total of 72 eyes were enrolled in the treatment experiment, 18 eyes in each group. There was no statistical difference in the clinical indexes on the 1st day after drug therapy. On the 5th day of treatment, the time of tear film rupture in group D was (2.933 卤0.320), () 2.900 卤0.280), (3.464 卤0.498) seconds and (3.643 卤0.413) seconds, respectively. The BUT of group C and group D were significantly longer than that of group A and group B, and the difference was statistically significant. Corneal fluorescein sodium staining grade was (11.640 卤1.008), (), 11.790 卤1.188), (10.330 卤1.371) and (10.270 卤1.104), respectively. Compared with the artificial tear group and the blank control group, the grade of corneal fluorescein sodium staining in the combined treatment group was significantly lower than that in the artificial tear group and the blank control group (F _ (6.145) P ~ (0. 001). The scores of corneal inflammation index in group A were (0.232 卤0.059), (, 0.229 卤0.078), (, 0.151 卤0.055) and (0.154 卤0.056), respectively. The inflammatory index of the two combined groups was significantly lower than that of the artificial tear group and the blank control group (F = 6.703, P < 0.001). There was no significant difference in the amount of basal tear secretion among the groups. There was no statistical difference in tear film rupture time, corneal fluorescein sodium staining and ocular surface inflammation between group C and group D. HE results showed that the thickness of epithelium in the blank control group was higher than that in the combined treatment group, and the epithelium was regular in the combined treatment group. PAS staining showed that the number of goblet cells in Praprofen group and flumilon group was similar, which was more than that in artificial tear group and blank control group. K10 was almost not expressed in the corneal epithelium of the Praprofen group and flumilon group, but remained positive in the artificial tear group and the blank control group. The level of TNF- 伪 in cornea decreased in both Praprofen group and flumilon group. Conclusion Sodium hyaluronate combined with Praprofen or flumilon has similar effects on dry eye of mice induced by benzalkonium chloride. Compared with sodium hyaluronate alone, sodium hyaluronate can stabilize tear film, promote epithelial repair and down-regulate the inflammatory factor TNF- 伪. Increase the number of goblet cells and inhibit squamous metaplasia. Praprofen can inhibit the inflammation of dry eye surface induced by benzalkonium chloride in mice and may be used in the treatment of moderate and severe dry eye.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R777
【参考文献】
相关期刊论文 前1条
1 崔国栋;苏金良;邱红;李蕾;宿可昕;;普拉洛芬联合人工泪液在干眼症中的应用[J];国际眼科杂志;2010年04期
，本文编号：2331527
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