中华眼视光学与视觉科学杂志
中華眼視光學與視覺科學雜誌
중화안시광학여시각과학잡지
CHINESE JOURNAL OF OPTOMETRY OPHTHALMOLOGY AND VISUAL SCIENCE
2012年
4期
197-202
,共6页
何欢%刘祖国%肖辛野%林志荣%周彤%何卉
何歡%劉祖國%肖辛野%林誌榮%週彤%何卉
하환%류조국%초신야%림지영%주동%하훼
干眼病%维生素A%鳞状上皮化生%杯状细胞%模型,动物,小鼠
榦眼病%維生素A%鱗狀上皮化生%杯狀細胞%模型,動物,小鼠
간안병%유생소A%린상상피화생%배상세포%모형,동물,소서
Xerophthalmia%Vitamin A%Metaplasia,squamous%Goblet cell%Models,animal,mice
目的 通过随机对照实验探讨维生素A对苯扎氯铵诱导的小鼠干眼的治疗作用.方法 实验研究.选用BALB/c小鼠50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(BUT)、角膜荧光素钠染色、炎症指数等参数选择干眼表现程度相近的小鼠30只(60眼)并随机分为A、B、C三组,每组10只(20眼).A、B组分别使用5000 IU/(g·ml)维生素A微乳剂和空白微乳剂滴眼,C组为空白对照组.在治疗的第0、1、3、5、7天检测BUT和角膜上皮荧光素钠染色、眼表炎症程度及基础泪液分泌量.7d治疗结束后取出小鼠眼球,进行HE及过碘酸-希夫(PAS)染色,角蛋白10(K10)免疫荧光标记,以及Western Blot检测角膜组织中K10表达水平.对数据采用单因素方差分析.结果 在药物治疗的第0、l、3、5天各项临床指标差异无统计学意义.治疗第7天时,A、B、C组BUT值分别为(4.20±0.89)s、(3.32±1.01)s和(3.28±0.74)s；其中A组的BUT值较B组和C组长,差异有统计学意义(F=6.825,P=0.002).A、B、C组角膜荧光素钠染色评分分别为3.74±1.31、5.47±1.81和5.83±1.54,其中维生素A微乳组较空白微乳组和空白对照组角膜荧光素钠染色分级轻,差异具有统计学意义(F=8.853,P＜0.01).各组间的角膜炎症指数评分、基础泪液分泌量差异均无统计学意义.组织病理检查示维生素A组与空白微乳组和空白对照组比较,其角膜上皮形态较规整；PAS染色示维生素A组杯状细胞数量更接近正常小鼠.维生素A组的角膜上皮层几乎均不表达K10,而空白微乳组及空白对照组呈阳性表达.Western Blot检测示维生素A组角膜中的K10水平低于空白微乳组和空白对照组.结论 维生素A可以延长苯扎氯铵诱导的干眼小鼠的BUT,降低角膜上皮染色程度,抑制角膜上皮鳞状化生,并能增加结膜杯状细胞数量,有望用于干眼治疗.
目的 通過隨機對照實驗探討維生素A對苯扎氯銨誘導的小鼠榦眼的治療作用.方法 實驗研究.選用BALB/c小鼠50隻,使用0.25％的苯扎氯銨溶液跼部點眼以誘導榦眼.在第21天時根據淚膜破裂時間(BUT)、角膜熒光素鈉染色、炎癥指數等參數選擇榦眼錶現程度相近的小鼠30隻(60眼)併隨機分為A、B、C三組,每組10隻(20眼).A、B組分彆使用5000 IU/(g·ml)維生素A微乳劑和空白微乳劑滴眼,C組為空白對照組.在治療的第0、1、3、5、7天檢測BUT和角膜上皮熒光素鈉染色、眼錶炎癥程度及基礎淚液分泌量.7d治療結束後取齣小鼠眼毬,進行HE及過碘痠-希伕(PAS)染色,角蛋白10(K10)免疫熒光標記,以及Western Blot檢測角膜組織中K10錶達水平.對數據採用單因素方差分析.結果 在藥物治療的第0、l、3、5天各項臨床指標差異無統計學意義.治療第7天時,A、B、C組BUT值分彆為(4.20±0.89)s、(3.32±1.01)s和(3.28±0.74)s；其中A組的BUT值較B組和C組長,差異有統計學意義(F=6.825,P=0.002).A、B、C組角膜熒光素鈉染色評分分彆為3.74±1.31、5.47±1.81和5.83±1.54,其中維生素A微乳組較空白微乳組和空白對照組角膜熒光素鈉染色分級輕,差異具有統計學意義(F=8.853,P＜0.01).各組間的角膜炎癥指數評分、基礎淚液分泌量差異均無統計學意義.組織病理檢查示維生素A組與空白微乳組和空白對照組比較,其角膜上皮形態較規整；PAS染色示維生素A組杯狀細胞數量更接近正常小鼠.維生素A組的角膜上皮層幾乎均不錶達K10,而空白微乳組及空白對照組呈暘性錶達.Western Blot檢測示維生素A組角膜中的K10水平低于空白微乳組和空白對照組.結論 維生素A可以延長苯扎氯銨誘導的榦眼小鼠的BUT,降低角膜上皮染色程度,抑製角膜上皮鱗狀化生,併能增加結膜杯狀細胞數量,有望用于榦眼治療.
목적 통과수궤대조실험탐토유생소A대분찰록안유도적소서간안적치료작용.방법 실험연구.선용BALB/c소서50지,사용0.25％적분찰록안용액국부점안이유도간안.재제21천시근거루막파렬시간(BUT)、각막형광소납염색、염증지수등삼수선택간안표현정도상근적소서30지(60안)병수궤분위A、B、C삼조,매조10지(20안).A、B조분별사용5000 IU/(g·ml)유생소A미유제화공백미유제적안,C조위공백대조조.재치료적제0、1、3、5、7천검측BUT화각막상피형광소납염색、안표염증정도급기출루액분비량.7d치료결속후취출소서안구,진행HE급과전산-희부(PAS)염색,각단백10(K10)면역형광표기,이급Western Blot검측각막조직중K10표체수평.대수거채용단인소방차분석.결과 재약물치료적제0、l、3、5천각항림상지표차이무통계학의의.치료제7천시,A、B、C조BUT치분별위(4.20±0.89)s、(3.32±1.01)s화(3.28±0.74)s；기중A조적BUT치교B조화C조장,차이유통계학의의(F=6.825,P=0.002).A、B、C조각막형광소납염색평분분별위3.74±1.31、5.47±1.81화5.83±1.54,기중유생소A미유조교공백미유조화공백대조조각막형광소납염색분급경,차이구유통계학의의(F=8.853,P＜0.01).각조간적각막염증지수평분、기출루액분비량차이균무통계학의의.조직병리검사시유생소A조여공백미유조화공백대조조비교,기각막상피형태교규정；PAS염색시유생소A조배상세포수량경접근정상소서.유생소A조적각막상피층궤호균불표체K10,이공백미유조급공백대조조정양성표체.Western Blot검측시유생소A조각막중적K10수평저우공백미유조화공백대조조.결론 유생소A가이연장분찰록안유도적간안소서적BUT,강저각막상피염색정도,억제각막상피린상화생,병능증가결막배상세포수량,유망용우간안치료.
Objective To investigate the therapeutic effect of Vitamin A on the recovery of dry eye that was induced by benzalkonium chloride (BAC) topical medication in the mouse.Methods In this randomized controlled experimental study,50 BALB/c mice were treated with the topical administration of 0.25％ BAC to establish the dry eye condition.Based on the consistency of tear film break-up time (BUT),corneal fluorescein staining scores and inflammation index,the eyes were re-selected and 30 mice were randomly divided into three groups on day 21 after the BAC treatment,with 10 mice (20 eyes) in each group.Group C was set up as a blank control,while groups A and B were treated with 5000 IU/(g·ml) Vitamin A eye drops and microemulsion solvent drops,respectively.BUTs,corneal fluorescein staining scores,inflammation index and tear volumes were evaluated in each group on days 0,1,3,5 and 7 after the therapeutic treatment.Global specimens were collected on day 7.Sections were stained with hematoxylin and eosin (HE) or by periodic acid-Schiff (PAS) assay,and labeled with cytokeratin 10 (K10) antibody.The expression of K10 in the cornea and conjunctiva was quantified by Western Blot.The data were analyzed by one-way ANOVAs.Results On days 0,1,3 and 5,no clinical differences were observed among the groups.On day 7,the BUT values were (4.20±0.89)s,(3.32±1.01)s and (3.28±0.74)s in groups A,B and C,respectively.The BUT in group A was significantly longer than those in groups B and C (F=6.852,P=0.002).The corneal fluorescein staining scores were 3.74±1.31,5.47±1.81 and 5.83±1.54 in groups A,B and C,respectively.The scores in group A were significantly lower than those in groups B and C (F=8.853,P＜0.01).No significant differences were found in the corneal inflammatory index or tear volume among the groups.Corneal morphology showed a loosened corneal epithelium feature in groups B and C,but comeal integrity in group A.PAS assay revealed similar goblet cell numbers between group A and normal eyes,but fewer goblet cells in groups B and C.Cytokeratin 10 was almost negatively expressed in Vitamin A treated groups,while it remained positive in the entire corneal epithelial layer with other treatments.The level of K10 in the cornea was down-regulated in Vitamin A treated groups.Conclusion Vitamin A treatment presented marked therapeutic effects on BAC-induced mouse dry eye,with a more stable tear film,better recovery of epithelial integrity,an increased number of goblet cells,and the elimination of squamous metaplasia,when compared to the treatment of microemulsion solvent eye drops only or without treatment.This experiment show the great potential of Vitamin A in the clinical treatment of dry eye.
