中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2013年
4期
339-346
,共8页
干燥综合征%干眼%C57BL/6.NOD-Aec1Aec2小鼠%组织病理学%炎症
榦燥綜閤徵%榦眼%C57BL/6.NOD-Aec1Aec2小鼠%組織病理學%炎癥
간조종합정%간안%C57BL/6.NOD-Aec1Aec2소서%조직병이학%염증
Sj(o)gren syndrome%Dry eye%C57BL/6.NOD-Aec1Aec2 mouse%Histopathology%Inflammation
背景 C57B L/6.NOD-Aec1 Aec2小鼠是研究人干燥综合征发病机制较理想的动物模型,但其干眼体征、眼表组织的病理学特点尚不清楚. 目的 观察C57 BL/6.NOD-Aec1 Aec2干燥综合征模型小鼠眼表组织的病理学变化,明确其是否能够自发地产生干眼. 方法 实验分为模型组(45只C57BL/6.NOD-Aec1 Aec2干燥综合征模型小鼠)和对照组(45只C57BL/6J小鼠).在两组小鼠第4、8、12、16、20周龄时依次进行空腹血糖检测、泪液分泌试验(SⅠt)、角膜和结膜丽丝胺绿染色及评分;5个时间点每组各取5只小鼠处死、取材,进行中央角膜上皮厚度测量、结膜杯状细胞计数,泪腺淋巴细胞浸润评估及角膜上皮细胞表面微绒毛观察. 结果 第4周龄时,两种小鼠均未见明显干眼体征.随着周龄的增加,C57BL/6.NOD-Aec1 Aec2小鼠泪液分泌逐渐减少,第8、12、16、20周龄酚红棉线长度依次为(2.7±0.9)、(2.5±0.8)、(1.8±0.6)、(1.9±0.1)mm,均明显短于同龄的C57BL/6J小鼠,差异均有统计学意义(P＜0.01);C57BL/6.NOD-Aec1 Aec2小鼠角膜结膜丽丝胺绿染色评分值随周龄增加明显升高,12、16和20周龄时其角膜结膜丽丝胺绿染色评分值均明显高于同龄C57BL/6J小鼠,差异均有统计学意义(P＜0.01).C57BL/6.NOD-Aec1 Aec2小鼠角膜上皮厚度值随周龄的增加逐渐降低,8、12、16、20周龄小鼠中央角膜上皮厚度值均明显低于同龄C57BL/6J小鼠,差异均有统计学意义(P＜0.01).C57BL/6.NOD-Aec1 Aec2小鼠结膜杯状细胞数随周龄的增加逐渐减少,8、12、16、20周龄时分别为(8.2±2.4)、(6.2±2.1)、(6.1±2.2)、(4.1±2.0)个,均明显少于同龄C57BL/6J小鼠,差异均有统计学意义(P＜0.01).泪腺淋巴细胞浸润及腺体破坏加重;扫描电子显微镜观察显示,角膜上皮细胞脱落增多,微绒毛减少,细胞紧密连接减少或中断;至第16周龄时,中央角膜上皮细胞完全脱落,可见内层排列规则的基质层胶原纤维.两种小鼠在第4、8、12、16、20周龄时的空腹血糖值均＜6.0 mmol/L,相同周龄时2个组间比较差异均无统计学意义(P=0.637、0.610、0.163、0.086、0.938),而C57BL/6J小鼠的泪液分泌值、中央角膜上皮厚度值、结膜杯状细胞数目及角膜上皮细胞表面的微绒毛密度等干眼指标均未随着小鼠周龄的增加而有所减少. 结论 C57BL/6.NOD-Aec1 Aec2干燥综合征模型小鼠可自发地产生干眼,且随着周龄的增加,其干眼体征加重.该模型小鼠可以较好地模拟人的干眼进程,可作为研究干眼发病机制的理想动物模型.
揹景 C57B L/6.NOD-Aec1 Aec2小鼠是研究人榦燥綜閤徵髮病機製較理想的動物模型,但其榦眼體徵、眼錶組織的病理學特點尚不清楚. 目的 觀察C57 BL/6.NOD-Aec1 Aec2榦燥綜閤徵模型小鼠眼錶組織的病理學變化,明確其是否能夠自髮地產生榦眼. 方法 實驗分為模型組(45隻C57BL/6.NOD-Aec1 Aec2榦燥綜閤徵模型小鼠)和對照組(45隻C57BL/6J小鼠).在兩組小鼠第4、8、12、16、20週齡時依次進行空腹血糖檢測、淚液分泌試驗(SⅠt)、角膜和結膜麗絲胺綠染色及評分;5箇時間點每組各取5隻小鼠處死、取材,進行中央角膜上皮厚度測量、結膜杯狀細胞計數,淚腺淋巴細胞浸潤評估及角膜上皮細胞錶麵微絨毛觀察. 結果 第4週齡時,兩種小鼠均未見明顯榦眼體徵.隨著週齡的增加,C57BL/6.NOD-Aec1 Aec2小鼠淚液分泌逐漸減少,第8、12、16、20週齡酚紅棉線長度依次為(2.7±0.9)、(2.5±0.8)、(1.8±0.6)、(1.9±0.1)mm,均明顯短于同齡的C57BL/6J小鼠,差異均有統計學意義(P＜0.01);C57BL/6.NOD-Aec1 Aec2小鼠角膜結膜麗絲胺綠染色評分值隨週齡增加明顯升高,12、16和20週齡時其角膜結膜麗絲胺綠染色評分值均明顯高于同齡C57BL/6J小鼠,差異均有統計學意義(P＜0.01).C57BL/6.NOD-Aec1 Aec2小鼠角膜上皮厚度值隨週齡的增加逐漸降低,8、12、16、20週齡小鼠中央角膜上皮厚度值均明顯低于同齡C57BL/6J小鼠,差異均有統計學意義(P＜0.01).C57BL/6.NOD-Aec1 Aec2小鼠結膜杯狀細胞數隨週齡的增加逐漸減少,8、12、16、20週齡時分彆為(8.2±2.4)、(6.2±2.1)、(6.1±2.2)、(4.1±2.0)箇,均明顯少于同齡C57BL/6J小鼠,差異均有統計學意義(P＜0.01).淚腺淋巴細胞浸潤及腺體破壞加重;掃描電子顯微鏡觀察顯示,角膜上皮細胞脫落增多,微絨毛減少,細胞緊密連接減少或中斷;至第16週齡時,中央角膜上皮細胞完全脫落,可見內層排列規則的基質層膠原纖維.兩種小鼠在第4、8、12、16、20週齡時的空腹血糖值均＜6.0 mmol/L,相同週齡時2箇組間比較差異均無統計學意義(P=0.637、0.610、0.163、0.086、0.938),而C57BL/6J小鼠的淚液分泌值、中央角膜上皮厚度值、結膜杯狀細胞數目及角膜上皮細胞錶麵的微絨毛密度等榦眼指標均未隨著小鼠週齡的增加而有所減少. 結論 C57BL/6.NOD-Aec1 Aec2榦燥綜閤徵模型小鼠可自髮地產生榦眼,且隨著週齡的增加,其榦眼體徵加重.該模型小鼠可以較好地模擬人的榦眼進程,可作為研究榦眼髮病機製的理想動物模型.
배경 C57B L/6.NOD-Aec1 Aec2소서시연구인간조종합정발병궤제교이상적동물모형,단기간안체정、안표조직적병이학특점상불청초. 목적 관찰C57 BL/6.NOD-Aec1 Aec2간조종합정모형소서안표조직적병이학변화,명학기시부능구자발지산생간안. 방법 실험분위모형조(45지C57BL/6.NOD-Aec1 Aec2간조종합정모형소서)화대조조(45지C57BL/6J소서).재량조소서제4、8、12、16、20주령시의차진행공복혈당검측、루액분비시험(SⅠt)、각막화결막려사알록염색급평분;5개시간점매조각취5지소서처사、취재,진행중앙각막상피후도측량、결막배상세포계수,루선림파세포침윤평고급각막상피세포표면미융모관찰. 결과 제4주령시,량충소서균미견명현간안체정.수착주령적증가,C57BL/6.NOD-Aec1 Aec2소서루액분비축점감소,제8、12、16、20주령분홍면선장도의차위(2.7±0.9)、(2.5±0.8)、(1.8±0.6)、(1.9±0.1)mm,균명현단우동령적C57BL/6J소서,차이균유통계학의의(P＜0.01);C57BL/6.NOD-Aec1 Aec2소서각막결막려사알록염색평분치수주령증가명현승고,12、16화20주령시기각막결막려사알록염색평분치균명현고우동령C57BL/6J소서,차이균유통계학의의(P＜0.01).C57BL/6.NOD-Aec1 Aec2소서각막상피후도치수주령적증가축점강저,8、12、16、20주령소서중앙각막상피후도치균명현저우동령C57BL/6J소서,차이균유통계학의의(P＜0.01).C57BL/6.NOD-Aec1 Aec2소서결막배상세포수수주령적증가축점감소,8、12、16、20주령시분별위(8.2±2.4)、(6.2±2.1)、(6.1±2.2)、(4.1±2.0)개,균명현소우동령C57BL/6J소서,차이균유통계학의의(P＜0.01).루선림파세포침윤급선체파배가중;소묘전자현미경관찰현시,각막상피세포탈락증다,미융모감소,세포긴밀련접감소혹중단;지제16주령시,중앙각막상피세포완전탈락,가견내층배렬규칙적기질층효원섬유.량충소서재제4、8、12、16、20주령시적공복혈당치균＜6.0 mmol/L,상동주령시2개조간비교차이균무통계학의의(P=0.637、0.610、0.163、0.086、0.938),이C57BL/6J소서적루액분비치、중앙각막상피후도치、결막배상세포수목급각막상피세포표면적미융모밀도등간안지표균미수착소서주령적증가이유소감소. 결론 C57BL/6.NOD-Aec1 Aec2간조종합정모형소서가자발지산생간안,차수착주령적증가,기간안체정가중.해모형소서가이교호지모의인적간안진정,가작위연구간안발병궤제적이상동물모형.
Background C57BL/6.NOD-Aec1Aec2 mouse is considered to be an idea model for the study of the pathogenesis of Sj(o)gren syndrome,but the cause of dry eye in these mice is unclear.Objective This study was to investigate the histopathological change of the ocular surfaces of C57BL/6.NOD-Aec1Aec2 mice,and to determine whether dry eye is developed spontaneously in these mice.Methods Forty-five clean C57BL/6.NOD-Aec1 Aec2 mice were used as the experiment group and forty-five C57BL/6J mice(both male and female)were used as the control group in this study.Detection of fasting blood-glucose,Schirmer' s test Ⅰ (S Ⅰ t),lissamine green staining and scoring of the corneal and conjunctival epithelium in the mice were performed at the age of 4,8,12,16 and 20weeks.Five mice from each group were sacrificed and their corneas were obtained to measure the central corneal epithelium thickness and to count the number of conjunctival goblet cells.In addition,lymphocyte infiltration in the lacrimal gland of the mice was examined with hematoxylin-eosin staining.The uhrastructure of the corneal epithelial cells and microvilli were assessed by scanning electron microscopy.The use and care of the mice were approved by the Experimental Animal Care Committee of the Third Military Medical University.Results No sign of dry eye was seen in both the 4-week-old C57BL/6.NOD-Aec1Aec2 mice and 4-week-old C57BL/6J mice.The S Ⅰ t values in 8-week-old,12-week-old,16-week-old and 20 week-old mice from the experiment group were (2.7 ±0.9) mm,(2.5 ±0.8) mm,(1.8±0.6) mm and (1.9± 0.1) mm,respectively,showing a significant reduction in comparison with those of the control mice of the same age(all P＜0.01).The amount of lissamine green staining in the C57BL/6.NOD-Aec1Aec2 mice gradually increased with age,showing elevated scores in 12-week-old,16-week-old and 20-week-old mice in the experiment group(all P＜0.01).The central corneal epithelium thicknesses were(20.18±3.75)μm,(17.01 ±5.25) μm,(14.19±5.72) μm and(12.00±3.25) μm in the 8-week-old,12-week-old,16-week-old and 20-week-old C57BL/6.NOD-Aec1Aec2 mice,respectively,which were significantly lower than those of the C57BL/6Jmice of the same age (all P＜0.01).The numbers of conjunctival goblet cells were (8.2±2.4),(6.2±2.1),(6.1 ±2.2) and (4.1 ± 2.0) in the 8-week-old,12-week-old,16-week-old and 20-week-old C57 BL/6.NOD-Aec 1Aec2 mice,respectively,showing a gradual decrease with age and a significant decline in comparison with those of the C57BL/6Jmice of the same age(all P＜0.01).Lymphocyte infiltration in the lacrimal gland and destruction of gland ducts were seen by hematoxylin-eosin staining,and acinar abnormality aggravated with aging.Reduction of corneal epithelial cells and the number of microvilli were distinguished with aging under the scanning electron microscope.The fasting bloodglucose levels of the two groups were both less than 6.0 mmol/L,and no significant difference was found between them at any age(P=0.637,0.610,0.163,0.086,0.938).Conclusions C57BL/6.NOD-Aec1Aec2 mice develop dry eye spontaneously with aging.The course of disease and characteristics of dry eye in C57BL/6.NOD-Aec1Aec2mice is similar to human dry eye.The C57BL/6NOD-Aec1 Aec2 mouse is the perfect model to study the pathogenesis of dry eye.
