论文部分内容阅读
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。
OBJECTIVE: To separate and detect the protein composition of rabbit aqueous humor by capillary zone electrophoresis (CZE) with high performance capillary electrophoresis (HPCE) and observe the effect of aqueous humor on cultured corneal stromal cells. METHODS: CZE was used to determine the protein content of New Zealand white rabbit aqueous humor: running buffer sodium borate buffer solution (pH = 8.7, 9.1, 9.4), capillary column temperature 20 ℃, detection wavelength 200 nm, Time 10s; operating voltage 20 kV. The first generation cultured corneal stromal cells were cultured in DMEM / F12, added with 10% aqueous humor. CCK-8 was used to detect the cell proliferation 24 hours later. The cell growth was observed under inverted microscope. Results: The capillary electrophoresis of rabbit aqueous humor with better separation efficiency was obtained by using sodium borate-borate solution with pH = 9.4. CCK8 absorbance increased (P <0.05) 24 h after adding aqueous humor, corneal stromal cells grew well. CONCLUSION: HPCE can rapidly and effectively detect rabbit aqueous humor protein. The rabbit aqueous humor with 10% volume fraction can promote the growth and proliferation of cultured rabbit corneal stromal cells.