普拉洛芬治療苯扎氯銨誘導的小鼠干眼的實驗研究
發(fā)布時間：2018-11-14 15:10
【摘要】：目的通過隨機對照實驗探討普拉洛芬對苯扎氯銨誘導小鼠干眼的治療作用及其可能機制。 方法選用BALB/c雄性小鼠70只,使用0.25%的苯扎氯銨溶液局部點眼以誘導干眼。在第21天時根據(jù)淚膜破裂時間、角膜熒光素鈉染色、炎癥指數(shù)等參數(shù)選擇干眼表現(xiàn)程度相似的眼并隨機分為A、B、C、D四組。A組為空白對照,B、C、D組分別使用0.1%玻璃酸鈉滴眼液,0.1%玻璃酸鈉滴眼液加0.1%氟米龍滴眼液,0.1%玻璃酸鈉滴眼液加0.1%普拉洛芬滴眼液進行治療。治療后第0、1、3、5天進行淚膜破裂時間和角膜上皮熒光素鈉染色,眼表炎癥程度的評估及基礎淚液量的測量。治療結束后取下小鼠眼球,進行HE及過碘酸-希夫(periodic acid-schiff, PAS)染色,角蛋白10(cytokeratin 10, K10)免疫熒光標記,以及Western blot檢測角結膜組織中腫瘤壞死因子α(tumor necrosis factor-α, TNF-α)表達水平。 結果共72眼入選治療實驗,每組18眼。在使用藥物治療后的第0、1、3天各項臨床指標無統(tǒng)計學差異。治療第5天時,A、B、C、D組淚膜破裂時間(BUT)分別為(2.933±0.320)、(2.900±0.280)、(3.464±0.498)和(3.643±0.413)秒;其中C組與D組的BUT均較A組和B組延長,差異具統(tǒng)計學意義(F=13.774,P=0.000)。角膜熒光素鈉染色分級A、B、C、D組分別為(11.640±1.008)、(11.790±1.188)、(10.330±1.371)及(10.270±1.104)分;兩聯(lián)合用藥治療組均較人工淚液組和空白對照組角膜熒光素鈉染色分級減輕,具有統(tǒng)計學意義(F=6.145,P=0.001)。角膜炎癥指數(shù)評分A、B、C、D組分別為(0.232±0.059)、(0.229±0.078)、(0.151±0.055)及(0.154±0.056)分;兩聯(lián)合用藥治療組均較人工淚液組和空白對照組炎癥指數(shù)降低,具有統(tǒng)計學意義(F=6.703,P=0.001)。各組間的基礎淚液分泌量無顯著性差異;C組與D組在淚膜破裂時間、角膜熒光素鈉染色分級及眼表炎癥程度中均無統(tǒng)計差異。HE結果顯示空白對照組上皮厚度高于兩聯(lián)合用藥組,聯(lián)合用藥組上皮規(guī)整;PAS染色示普拉洛芬組及氟米龍組的杯狀細胞數(shù)量相當,均多于人工淚液組及空白對照組。普拉洛芬組及氟米龍組的角膜上皮層均幾乎不表達K10,而人工淚液組及空白對照組仍陽性表達。普拉洛芬組及氟米龍?zhí)幚斫M角膜中的TNF-α水平均有下降。 結論玻璃酸鈉聯(lián)合普拉洛芬或氟米龍對苯扎氯銨誘導的小鼠干眼具有相似效果,與單用玻璃酸鈉相比,可更好地穩(wěn)定淚膜,促進上皮修復,下調(diào)炎癥因子TNF-α,增加杯狀細胞數(shù)量,抑制鱗狀上皮化生。普拉洛芬可以抑制苯扎氯銨誘導的小鼠干眼眼表炎癥,有望用于臨床中重度干眼的治療。
[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 偽 (tumor necrosis factor- 偽 in the cornea conjunctiva. TNF- 偽) 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 鹵0.320), () 2.900 鹵0.280), (3.464 鹵0.498) seconds and (3.643 鹵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 鹵1.008), (), 11.790 鹵1.188), (10.330 鹵1.371) and (10.270 鹵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 鹵0.059), (, 0.229 鹵0.078), (, 0.151 鹵0.055) and (0.154 鹵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- 偽 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- 偽. 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.
【學位授予單位】：南華大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R777
本文編號：2331528
[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 偽 (tumor necrosis factor- 偽 in the cornea conjunctiva. TNF- 偽) 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 鹵0.320), () 2.900 鹵0.280), (3.464 鹵0.498) seconds and (3.643 鹵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 鹵1.008), (), 11.790 鹵1.188), (10.330 鹵1.371) and (10.270 鹵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 鹵0.059), (, 0.229 鹵0.078), (, 0.151 鹵0.055) and (0.154 鹵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- 偽 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- 偽. 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.
【學位授予單位】：南華大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R777
【參考文獻】
相關期刊論文 前1條
1 崔國棟;蘇金良;邱紅;李蕾;宿可昕;;普拉洛芬聯(lián)合人工淚液在干眼癥中的應用[J];國際眼科雜志;2010年04期
，本文編號：2331528
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