目的：通过前房注射亚硒酸钠溶液建立新西兰兔硒性白内障模型，并评估验证其在药效试验中的应用。方法：采用前房注射10 mmol/L亚硒酸钠溶液(0.1 mL/只)对动物右眼进行造模，于造模第3天根据晶状体混浊度评分选取成模动物分为模型组、吡诺克辛滴眼液(pirenoxine sodium eye drops, PSED)0.05 mg/mL组，以模型组动物左眼作为空白组，每组8只眼；分组后对各组动物眼进行滴眼给药，50 µL/眼/次，3 次/天，连续给药17 d。于给药前、给药第7、17天进行裂隙灯检查拍照在体评估晶状体混浊度，末次给药后完整分离各组动物晶状体，体外评估晶状体透明度，然后将各组动物晶状体匀浆，用于过氧化物酶(peroxidase, POD)及谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-Px)活力测定。结果：与空白组相比，模型组给药前、给药第7、17天晶状体混浊度评分均升高，给药结束后晶状体透明度评分升高，晶状体中GSH-PX和POD活力均降低；与模型组相比，PSED 0.05 mg/mL组给药17d晶状体混浊度和透明度评分均降低，晶状体中GSH-PX活力升高，POD无变化。结论：前房注射亚硒酸钠溶液可诱导新西兰兔发展出稳定的白内障模型症状，适用于药物药效作用的评价。
Objective: To establish a selenium-induced cataract model in New Zealand rabbits by anterior chamber injection of sodium selenite solution and evaluate and verify its application in pharmacodynamic trials. Methods: The right eyes of the animals were modeled by anterior chamber injection of 10 mmol/L sodium selenite solution (0.1 mL per animal). On the third day of modeling, the modeled animals were selected and divided into the model group and the pirenoxine sodium eye drops (PSED) 0.05 mg/mL group based on the lens turbidity score. The left eyes of the animals in the model group were taken as the blank group, with 8 eyes in each group. After grouping, eye drops were administered to the eyes of each group of animals. The dosage was 50 µL per eye each time, three times a day, for 17 consecutive days. Slit lamp examination and photography were conducted on the 7th and 17th days of administration to evaluate the turbidity of the lenses in vivo. After the last administration, the lenses of each group of animals were completely isolated, and the transparency of the lenses was evaluated in vitro. Then, the lenses of each group of animals were homogenized. It is used for the determination of peroxidase (POD) and glutathione peroxidase (GSH-Px) activities. Results: Compared with the blank group, the lens turbidity scores of the model group were significantly increased before administration, on the 7th and 17th days of administration, the lens transparency scores were significantly increased after the end of administration, and the activities of GSH-PX and POD in the lens were significantly decreased. Compared with the model group, the scores of lens turbidity and transparency in the PSED 0.05 mg/mL group and 17 days after administration were significantly decreased, the activity of GSH-PX in the lens was significantly increased, and there was no change in POD. Conclusions: Anterior chamber injection of sodium selenite solution can induce the development of stable cataract model symptoms in New Zealand rabbits and is suitable for the screening and evaluation of drug efficacy.
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.
