中国实用眼科杂志
中國實用眼科雜誌
중국실용안과잡지
CHINESE JOURNAL OF PRACTICAL OPHTHALMOLOGY
2011年
11期
1203-1206
,共4页
JAM-1%组织培养保存%活性染色%透射电子显微镜
JAM-1%組織培養保存%活性染色%透射電子顯微鏡
JAM-1%조직배양보존%활성염색%투사전자현미경
JAM-1%Tissue culture preservation%Vital staining%Transmission electron microscopy
目的 通过阻断接合粘附分子-l引起角膜内皮细胞结构和活性的改变来研究JAM-1的作用.方法 由14只大鼠取下28片3.5mm直径角膜片,其中右眼角膜为实验组,用抗JAM-1单克隆抗体处理,左眼角膜为对照组,用磷酸缓冲液处理.处理后在高糖DMEM液中进行培养保存5d,之后用含5％葡聚糖T500的DMEM脱水24 h.脱水后每组有12片角膜做锥蓝-茜素红染色内皮细胞活性计数及厚度测量,2片角膜与另外2片新鲜角膜一同进行戊二醛固定,用于透射电子显微镜观察.结果 活性细胞计数对照组为(2008±505 )/mm2,实验组为(934±521 )/mm2,P＜0.0l.角膜厚度对照组为(375.02±83.33)μm,实验组为(461.81±39.14)μm,P＜0.01.超微结构显示,实验组内皮细胞损坏严重,有较多的自噬体形成.结论 作为细胞紧密连接构成成份的JAM-1,对维持角膜的内皮细胞结构和活性必不可少.
目的 通過阻斷接閤粘附分子-l引起角膜內皮細胞結構和活性的改變來研究JAM-1的作用.方法 由14隻大鼠取下28片3.5mm直徑角膜片,其中右眼角膜為實驗組,用抗JAM-1單剋隆抗體處理,左眼角膜為對照組,用燐痠緩遲液處理.處理後在高糖DMEM液中進行培養保存5d,之後用含5％葡聚糖T500的DMEM脫水24 h.脫水後每組有12片角膜做錐藍-茜素紅染色內皮細胞活性計數及厚度測量,2片角膜與另外2片新鮮角膜一同進行戊二醛固定,用于透射電子顯微鏡觀察.結果 活性細胞計數對照組為(2008±505 )/mm2,實驗組為(934±521 )/mm2,P＜0.0l.角膜厚度對照組為(375.02±83.33)μm,實驗組為(461.81±39.14)μm,P＜0.01.超微結構顯示,實驗組內皮細胞損壞嚴重,有較多的自噬體形成.結論 作為細胞緊密連接構成成份的JAM-1,對維持角膜的內皮細胞結構和活性必不可少.
목적 통과조단접합점부분자-l인기각막내피세포결구화활성적개변래연구JAM-1적작용.방법 유14지대서취하28편3.5mm직경각막편,기중우안각막위실험조,용항JAM-1단극륭항체처리,좌안각막위대조조,용린산완충액처리.처리후재고당DMEM액중진행배양보존5d,지후용함5％포취당T500적DMEM탈수24 h.탈수후매조유12편각막주추람-천소홍염색내피세포활성계수급후도측량,2편각막여령외2편신선각막일동진행무이철고정,용우투사전자현미경관찰.결과 활성세포계수대조조위(2008±505 )/mm2,실험조위(934±521 )/mm2,P＜0.0l.각막후도대조조위(375.02±83.33)μm,실험조위(461.81±39.14)μm,P＜0.01.초미결구현시,실험조내피세포손배엄중,유교다적자서체형성.결론 작위세포긴밀련접구성성빈적JAM-1,대유지각막적내피세포결구화활성필불가소.
Objective To approach the function of JAM-1 in corneal endothelium by observing cornea endothelium structure and activity change induced by antibody blockage.Methods Twenty-eight corneas were obtained from 14 rats and were divided into two groups.Corneas of the right eye were in experiment group and processed with monoclonal antibody of JAM-1 to block the function of JAM-1; corneas of the left eye were in control group and processed with PBS buffer.All corneas were cultured in DMEM with high glucose for 5 days and then dehydrated in DMEM containing 5％dextran-T500 for 24 hours.Twelve corneas of each group were underwent Trypan blue - alizarin vital staining and cornea thickness were measured.Two corneas of each group and other 2 fresh normal corneas were fixed with 4％ glutaraldehyde for transmission electron microscopy.Results The active cell counting of experiment group and control group were (2008± 505)/mm2 and (934± 521 )/mm2 respectively,P ＜0.01.The cornea thickness of the two groups were (375.02± 83.33)μ m and (461.81±39.14)μ m respectively,P ＜0.01.Transmission electron microscopy showed that in experiment group the endothelium cell appeared more ultrastructural damage and autophagosome formation.Conclusions JAM-l,as a component of cell junctions,its function is essential for corneal endothelium structure and activity in tissue culture preservation.
