大鼠青光眼滤过术后滤过泡形态与组织病理学动态观察

Dynamic changes of morphology and histopathology of conjunctival filtering bleb after anti-glaucoma filteringsurgery in rat

背景制作合适的青光眼滤过术后球结膜滤过泡的动物模型并观察滤过泡形态及组织病理学的动态变化，有助于了解青光眼术后滤过泡瘢痕化的病理变化，并掌握青光眼术后抗滤过泡瘢痕化治疗的时间窗和治疗靶点。目的动态观察大鼠滤过术后球结膜滤过泡生存时间、形态学及组织学改变，了解前房引流管植入术后滤过泡的形成及瘢痕化进程。方法选用健康成年雄性SD大鼠60只，采用前房引流管植入法建立大鼠球结膜滤过泡模型。分别于术后1、3、5、7、14、28d处死大鼠，获得术眼结膜滤过泡组织，并在裂隙灯显微镜下动态观察滤过泡的变化。参照Kronfeld滤过泡形态及功能分型方法对滤过泡进行分型，标尺测量滤过泡的长度和宽度，观察术后不同时间滤过泡面积的变化。分别于上述时间点获取滤过区组织，采用苏木精一伊红染色动态观察术后滤过泡的组织病理学变化。结果所有术眼在术后第1天均形成不同程度隆起的滤过泡。滤过泡生存时间为7～17d，平均生存时间为（11．93±2．23）d；术后1～5d，滤过泡的生存率均为100％，术后7d降至77％，术后14d降至20％，术后28d功能性滤过泡全部消失。术后5d，均为功能性滤过泡，均为Ⅰ型；术后7d，23眼为功能性滤过泡，其中Ⅰ型13眼，Ⅱ型10眼；术后14d，4眼为功能性滤过泡，其中Ⅰ型2眼，Ⅱ型2眼。滤过泡面积由术后第1天的（7．50±1．08）iTlm。逐渐缩小为术后第14天的（1．83±0．35）mm。。术后不同时间点滤过泡面积整体比较，差异有统计学意义（F=180．056，P=0．000），不同时间点间两两比较差异均有统计学意义（均P=0．000）。组织病理学检查表明，术后1～3d滤过泡组织的病理改变以组织水肿、炎性细胞浸润、充血等炎症改变为主，自术后第5天，滤过泡组织成纤维细胞增加，随着时间延长，滤过泡组织以成纤维细胞、血管形成及胶原纤维形成等增生性表现为主。结论前房引流管植入术可成功建立大鼠球结膜滤过泡模型，术后炎症、纤维增生和胶原纤维形成参与滤过泡的瘢痕化过程。

Background Dynamic observation in morphology and histology of conjunctival filtering bleb in rats can contribute to form ideal filtering blebs by interferencing the process of scarring. Objective This study was designed to observe the survival time,morphological and histopathological changes of conjunetival filtering bleb after filtering surgery. Methods Filtering surgeries were performed on 60 male adult Sprague-Dawley rats by implanting drainage tubes into the anterior chamber. The shape of filtering blebs was examined by slit lamp microscope 1 day,3, 5,7,14,28 days after surgery. The filtering was typed according to the criteria of Kronfeld. Areas of the fihering bleb were measured and compared at various time points after surgery. Conjunctival and subconjunctival tissues at filtering zone were obtained 1 day,3,5,7,14,28 days after surgery, respectively, and hematoxylin and eosin staining was used for the histopathologieal evaluation. Results Well-elevated filtering blebs formed on the first day after surgery in 60 rat eyes. The survival time of filtering bleb was 7-17 days with the mean time of （11.93±2.23） days. Survivalrate of the filtering blebs was 100% in 1-5 days,77% in 7 days,20% in 14 days and 0 in 28 days after operation. The filtering blebs were functional （ Ⅰtype） in 1-5 days after operation, 23 eyes were functional in 7 days after operation （ 13 eyes wereⅠtype, 10 eyes were Ⅱ type） ,4 eyes were functional in 14 days after operation （2 eyes were I type,2 eyes were Ⅱ type）. The filtering area shrunken from （7.50± 1.08 ） mm2 to （ 1.83 ± 0.35 ） mm2 during the observational duration. The filtering blebs area of different days after surgery were statistically significant （F = 180. 056,P= 0. 000）, and significant differences were found between any two timepionts after surgery （all at P = 0. 000）. Histological examination showed that inflammatory finding was dominant in postoperative day 1-3, such as infiltration of inflammatory cells, vasodilatation and edema of tissue. However, proliferating manifestation became the main signs 5 days later,showing the increase of fibroblasts,neovascularization and collagen formation. Conclusions The model of conjunctival filtering blebs in rats can be successfully established by implanting silicone tube into anterior chamber. The inflammatory response, proliferation of flbroblasts and the accumulation of collagen fibers are involved in the scarring process of filtering blebs.