兔房水HPCE检测及其对培养角膜基质细胞的作用  

Aqueous humor constitutes of rabbit detected with HPCE and their effect on the growth of keratocytes

作　　者：陈建苏[1,2] 李晓霞[1,2] 唐光霞[1,2] 麦志伟[3] 王升威[4] 洪嘉凡[3] 罗子源[4] 李泽键[4] 卢春婷[3] 

机构地区：[1]暨南大学医学院眼科研究室,广东广州510632 [2]暨南大学再生医学教育部重点实验室,广东广州510632 [3]暨南大学医学院口腔医学系,广东广州510632 [4]暨南大学医学院临床医学系,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2010年第4期374-377,共4页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金项目(30371519;30973244)

摘　　要：目的:采用高效毛细管电泳(HPCE)的毛细管区带电泳(CZE)分离与检测兔房水中各蛋白组成,观察房水对培养角膜基质细胞的作用。方法:CZE测定新西兰白兔房水蛋白成分:运行缓冲液硼酸钠-硼酸缓冲溶液(pH=8.7、9.1、9.4);毛细管柱温20℃;检测波长200 nm;压力进样0.5 psi,进样时间10s;运行电压20 kV。第1代培养角膜基质细胞进行DMEM/F12培养,加入体积分数为10%的房水,CCK-8检测24 h后细胞增殖情况,每天倒置显微镜下观察细胞生长情况。结果:应用pH=9.4的硼酸钠-硼酸缓冲溶液可获得分离效果较理想的兔房水毛细管电泳峰图。加入房水24 h后CCK8检测吸光度增高(P<0.05),角膜基质细胞生长良好。结论:HPCE可快速有效方便检测兔房水蛋白成分,体积分数为10%的兔房水可促进培养的兔角膜基质细胞生长与增殖。Aim：To detect the protein constitutes of rabbit aqueous humor with capillary zone electrophoresis（CZE） of high performance capillary electrophoresis（HPCE） and the effect of aqueous humor on the growth of keratocytes. Methods： Samples of rabbit aqueous humors were analyzed using CZE.The running buffers were borate-borax（pH=8.7,9.1 and 9.4）.Sample application was 10s,applied voltage 20kV,detection at 200nm,sample pressure was 0.5psi and column temperature was maintained at constant 20°C.Rabbit keratocytes were cultured in vitro and 10% rabbit aqueous humors were added in the culture medium DMEM/F12.The cell proliferations after 24 hours were examined by the CCK-8 assay.Phase contrast microscope examination was performed daily.Results： A number of resolved peaks were obtained from aqueous humor of rabbit in running buffers of borate-borax（pH= 9.4）.The absorbance values of the CCK8 assay for supplementary aqueous humors after 24 hours were significantly high（P0.05）.Rabbit keratocytes grew well.Conclusion：CZE is rapid,effective and convenient for the detection of the protein constitutes of rabbit aqueous humor.10% aqueous humors promote the growth and proliferation of keratocytes.

关 键 词：房水 高效毛细管电泳(HPCE) 毛细管区带电泳(CZE) 角膜基质细胞 分离 

分 类 号：R331[医药卫生—人体生理学]