Purpose: To investigate the inhibitory effect of diclofenac sodium on rabbit corneal epithelial cells (RCECs) in vitro and explore its pharmacological mechanism.
Methods: The fresh rabbit cornea was cultured to get the primary RCECs, and RCECs of passage 2 were used for the research. The cells were divided into experimental groups, in which the cells were incubated with different concentrations (18.18, 27.27, 36.36, 45.45, 54.55 μg/ml) of diclofenac sodium, and a control group. The effect of diclofenac sodium on the proliferation of cells was measured by methyl thiazolyl tetrazolium (MTT) assay 24, 48, and 72 h after incubation. While the RCECs were divided into experimental groups, the cells in which were incubated with 9 and 12.5 μg / ml diclofenac sodium, and a control group. The cell cycle and apoptotic rate were observed by flow cytometer.
Results: MTT assay showed that diclofenac sodium had an obvious inhibitory effect on RCECs, and the inhibition rate was increasing along with the increase of the concentration of diclofenac sodium and the incubation time (P < 0.05). Flow cytometer showed that after incubation with diclofenac sodium, the cells in G0/G1 phase were obviously increased, and the apoptosis cusp and apoptotic rate were increased.
Conclusion: Diclofenac sodium has an obvious inhibitory effect on RCECs, which was dosage-dependent, and it may function by inducing cell apoptosis and ceasing cell cycles
Purpose: To investigate the inhibitory effect of diclofenac sodium on rabbit corneal epithelial cells (RCECs) in vitro and explore its pharmacological mechanism.
Methods: The fresh rabbit cornea was cultured to get the primary RCECs, and RCECs of passage 2 were used for the research. The cells were divided into experimental groups, in which the cells were incubated with different concentrations (18.18, 27.27, 36.36, 45.45, 54.55 μg/ml) of diclofenac sodium, and a control group. The effect of diclofenac sodium on the proliferation of cells was measured by methyl thiazolyl tetrazolium (MTT) assay 24, 48, and 72 h after incubation. While the RCECs were divided into experimental groups, the cells in which were incubated with 9 and 12.5 μg / ml diclofenac sodium, and a control group. The cell cycle and apoptotic rate were observed by flow cytometer.
Results: MTT assay showed that diclofenac sodium had an obvious inhibitory effect on RCECs, and the inhibition rate was increasing along with the increase of the concentration of diclofenac sodium and the incubation time (P < 0.05). Flow cytometer showed that after incubation with diclofenac sodium, the cells in G0/G1 phase were obviously increased, and the apoptosis cusp and apoptotic rate were increased.
Conclusion: Diclofenac sodium has an obvious inhibitory effect on RCECs, which was dosage-dependent, and it may function by inducing cell apoptosis and ceasing cell cycles
目的:通过观察对金黄色葡萄球菌性角膜溃疡的疗效,筛选克拉霉案眼用凝胶的合适浓度。方法:角膜实质层接种法建立40只家兔右眼金黄色葡萄球菌性角膜溃疡模型,将模型随机分成5组,每组8只免(8只眼),各组分别给予空白基质、0.1%克拉素眼用凝胶、0.25%克拉霉素眼用凝胶、左氧氟沙星凝胶、0.25%克拉霉素眼用凝胶联合重组牛碱性成纤维细胞生长因子(Recombinant bovine basic fibroblast growth factor, Rb-bFGF)，每天4次，每次2滴，分别在第1、3、5、7、10、14 天观察角膜病变情况及溃疡面积大小。结果:在相同的给药方法下，0.1%克拉霉素眼用凝胶、0.25%克拉素眼用凝胶、左氧沙星凝胶、0.25%克拉霉素眼用凝胶联合Rb-bFCF均能使金黄色葡萄球菌性角膜溃疡面积缩小角膜病变好转，与空白基质组相比有统计学差异(P < 0.05)：0.25%克拉素眼用凝胶组疗效明显优于0.1%克拉荐素眼用凝胶组(P < 0.05)。结论:制备的 0.25%克拉霉素眼用凝胶对金黄色葡萄球菌性角膜溃疡疗效肯定，可以进一步开发应用于临床。
Purpose:To screen proper concentration of clarithromycin ophthalmic gel by observingthe efficacy of different concertrations of clarithromycin ophthalmic gel for treatingstaphylococcal corneal ulcers.
Methods:Corneal ulcer was induced in the right eye of 40 rabbits, 3.0 x 10°CFU/mlstaphylococcus aureus suspension was injected midstromally into the central cornel.These rabbits were divided randomly into $ groups ,each group received respectivelytopical blank matrix, clarithromycin ophthalmic gel 0.1%, clarithromycin ophthalmicgel 0.25%,levofloxacin ophthalmic gel, clarithromycin ophthalmic gel 0.25% andrecombinant bovine basic fibroblast growth factor (Rb-bFGF), 4 times every day, 2drops each time. The eyes were examined respectively with the slit lamp beforetreatment(day0), on day3, day5, day7, day 10, day 14 to observe theprogression of corneal ulceration. including the area of the corneal ulcer and mark of keratitis.Resuls:Under the same way of giving medicine, experimental coreal ulcer studiesshowed a statistically significant decrease in all tratement groups on measurements ofthe area of the comeal ulcer and mark of keratitis(P<0.05), and clarithromycinophthalmic gel 0.25% had a better action than clarithromycin ophthalmic gel 0.1%against staphylococcus aureus corneal ulcer.
Conclusion:Clarithromycin ophthalmic gel 0,25% was proved to be an effective ocularmedication for the therapy of gram-positive bacterial corneal ulcer.
目的: 研究 TGF-β1 短期眼部应用对兔角膜碱烧伤后整合素 β1 表达和角膜上皮愈合的影响,探求其对角膜碱烧伤的治疗作用。方法: 制备大耳白家兔角膜碱烧伤模型, 一组给予 TGF-β1 (浓度为 200 ng /ml) 局部滴眼, 每日 3 次, 连续 7 日; 另一组给予 PBS 溶液代替, 处理相同。于角膜碱烧伤后每日观察角膜上皮愈合面积, 并于烧伤后 6 h、1 d、3 d、7 d 和 14 d 5 个时间点应用免疫组化方法检测 TGF-β1 实验组与 PBS 组角膜整合素 β1 表达情况。结果: 烧伤后 4 d、10 d、11 d、12 d 和 14 d 实验组和对照组上皮愈合率比较, 差异有统计学意义(P < 0.05) , 两组随着上皮修复过程的进行, 整合素 β1 的表达均逐渐增加, 烧伤后 7 d、14 d两个时间点实验组和对照组整合素 β1 平均灰度值比较, 差异有显著性(P < 0.05) 。结论: TGF-β1 在活体实验中能促进整合素 β1 的表达, 而后者的增加可以促进角膜上皮细胞向损伤区域的移行和粘附, 从而减少碱烧伤愈合过程中上皮再次脱落现象, 有利于创伤愈合。
Purpose: To observe the effect of TGF-β1 applied topically to the alkali-injured rabbit eye on corneal epithelial wound healing and expression of integrin β1 and its therapeutic action on corneal alkali burns.
Methods: Alkali burn was produced in 60 corneas from 30 rabbits. Two groups were randomly divided. One group was treated with TGF-β1 solution (200 ng /ml) topically 3 times one day within the first 7 days, the other group was treated with phosphate-buffered saline (PBS) solution. The injured eyes were photographed after the fluorescence staining with a digital camera and the pictures were analyzed with computer-aided picture analysis system to calculate the rate of corneal epithelial healing. The expression of integrin β1 was investigated in the point 6 h, 1 d, 3 d, 7 d, 14 d after the injury by means of immunohistochemical analysis.
Results: On the 4th, 10th, 11st, 12nd and 14th days after the burning, the rate of corneal epithelial healing of TGF-β1 groups was markedly higher than that of the PBS group (P < 0.05) . The expression of integrin β1 in the cornea epithelial cells gradually increased during the wound healing. On the 7th and 14th days after the burning, the expression of integrin β1 in the cornea epithelial cells of TGF-β1 group was remarkably higher than that of the PBS group(P < 0.05) .Conclusions: TGF-β1 could up-regulate integrin β1 in vivo corneal alkali burn model, which could stimulate the cornea epithelial cells to migrate and adhere to the cornea stroma, that can reduce the cases of the epithelial cells_detachment from the cornea stroma and sustain the corneal reepithelization.
