开放式压力控制培养系统对人眼小梁细胞超微结构及纤维连接蛋白合成的影响  

Effects of Pressure on Ultrastructure and Fibronectin Synthesis of Cultured Human Trabecular Meshwork Cells by Open Pressure Controlling Culture System

作　　者：张虹[1] 单长梅[1] 李贵刚[1] 陈莲一[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院眼科,武汉430030

出　　处：《华中科技大学学报（医学版）》2007年第4期510-513,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：湖北省自然科学基金资助项目(No.2003ABA150)

摘　　要：目的采用开放式压力控制培养系统,研究压力对人眼小梁细胞(HTMCs)超微结构及纤维连接蛋白(FN)合成的影响。方法组织块法体外培养HTMCs,选取第5代细胞分为实验组和对照组。对照组不施加压力,实验组采用开放式压力控制培养系统分别施加20、40、608、0 mmHg(1 mmHg=0.133 kPa)的压力,压力作用时间分别为12、24、36 h。在光学显微镜、透射电镜下观察HTMCs形态及超微结构的变化。采用免疫组化方法和图像分析法检测FN的表达变化。结果透射电镜显示:20 mmHg压力作用12、24、36 h后与对照组比较,HTMCs超微结构未见明显改变;40mmHg压力作用12、24、36 h后HTMCs胞质内出现大小不等的空泡;60、80 mmHg压力作用12、24、36 h后,HTMCs出现线粒体嵴变得短而少或者丧失、溶酶体髓样结构增多等不可逆性改变。免疫组化结果显示体外培养的HTMCs均阳性表达FN;图像分析显示:20 mmHg压力作用12、24、36 h后HTMCs合成FN与对照组比较差异均无显著性意义(均P>0.05);40 mmHg压力作用12、24、36 h后HTMCs合成FN明显增加,差异均有显著性意义(均P<0.01);而60、80 mmHg压力作用12、243、6 h后HTMCs合成FN则逐渐下降(均P<0.01),但与对照组比较仍有不同程度增高(均P<0.01)。结论环境压力升高可以引起小梁细胞形态、超微结构以及FN的表达发生变化,从而可能在原发性开角型青光眼的发生和发展过程中发挥重要作用。Objective To investigate the effects of pressure on ultrastructure and fibronectin （FN） synthesis of human trabecular meshwork cells （HTMCs） by open pressure controlling culture system. Methods By using tissue-culture method, HTMCs were cultured in vitro and those in 5th passage were randomly divided into control group and experimental groups. By open pressure controlling culture system the experimental groups were given 20, 40, 60 and 80 mmHg （1 mmHg=0. 133 kPa）, respectively, while no pressure was given in the control group. In each group, the pressure sustained for 12, 24 and 36 h, respectively. The morphologic and ultrastructural changes of HTMCs were observed under light/transmission electron microscopy. The FN expression was detected by immunocytochemistry and computer image analysis. Results Transmission electron microscopy revealed that there was no remarkable difference in the morphology and ultrastructure of HTMCs between the control group and 20 mmHg/12, 24, 36 h groups. Some HTMCs turned into vacuoles in cytoplasm, in 40 mmHg/12, 24, 36 h groups. Mitochondrial crista became shorter and fewer even lost, and some irreversible changes could be observed. In the 60 and 80 mmHg/12, 24, 36 h groups, immunocytochemistry showed the strong FN expression staining in HTMCs. The image analysis showed that there were no significant differences in the FN synthesis between the control group and 20 mmHg/12, 24, 36 h groups （P〉0.05）, while between the control group and 40 mmHg/12, 24, 36 h groups, the difference was significant （P 〈0.01）. In the 60 and 80 mmHg groups, the FN synthesized by HTMCs was decreased, but still higher than in the control group. Conclusion The beget pressure setting could result in the changes in the morphology, ultrastructure and FN expression of HTMCs, and may play an important role in the occurrence and development of the primary open angle glaucana.

关 键 词：压力 纤维连接蛋白 小梁细胞 青光眼 

分 类 号：R775.2[医药卫生—眼科]