| Objective: Cataract is first in the world of blinding eye disease. Despite years of domestic and foreign scholars through a variety of methods to study, its pathogenesis has not been possible to completely clear. Cataract surgery at the same time improve the treatment of cataracts in the research have also made great progress, but has been unable to find a good control of the occurrence and development of cataract drugs, so countries continued exploration and research scholars. In recent years, the study found that cataract patients blood, aqueous and lens tear of nitric oxide (nitric oxide, NO) and nitric oxide synthase (nitric oxide synthase,NOS) content are increased. NO is a highly unstable free radical biology, molecular small, simple structure, rapid proliferation through the biofilm, biological half-life of only 3-5s, their generation depends on nitric oxide synthase, NO exist due to the short time available, drug it is very difficult for its direct effect, mostly by inhibiting nitric oxide synthase pathway inhibitor to study its generation. This experiment to be prepared through the galactose cataract model in rats, and nitric oxide synthase inhibitor - nitro-L-arginine methyl ester (L-NAME) eyedrops to intervene, slit-lamp microscope to observe the morphological changes of lens , nitric acid reductase method spectrophotometer determined detection of nitric oxide and nitric oxide synthase content and technical analysis by flow cytometry detection of apoptotic gene caspase-3 expression to explore the nitric oxide synthase inhibitor in rats galactose cataract role.Methods: 50 three-week-old Wistar rats, were equally divided between male and female, weight 50g; mydriasis with compound tropicamide eye drops and examination, no eye diseases, eye lens transparency. Randomly divided into 3 groups: A group: blank group10, no special treatment, fed with ordinary particle feed, water and food freely perturbation; B: model group 20, with the blank group feeding, daily morning subcutaneous injection of the concentration of the back neck for 50% of D-galactose solution 30m1/kg (ie :15g/kg/d), continuous injection for 30 days; C: treatment group, 20 am to deal with the model group, while L-NAME eye drops given 3 / day to the end of the experiment. Experimental section 10,20,30 days to Wistar rats in each group are given compound tropicamide eye drops mydriasis full line and slit-lamp microscopic examination, and score; At the thirty day of the experiment ,three groups animals are killed after femoral artery bleeding and taking their lens, half of the lens mechanism broken up into a tissue homogenate measured NO content and NOS; half of the lens with fixative fixed, flow cytometry to detect apoptotic gene caspase-3 content. Spss13.0 experimental data with statistical software for analysis. Using the normal distribution test, variance analysis of statistical data, statistical results are mean±standard deviation expressed (P <0.05 The results show there is significant difference).Results:1 Slitlamp observation of changes in rat lens in each group:1.1 Under the slit lamp camera lens, see the attached map;1.2 Degree of lens opacity score such as Yang Bin reported in accordance with the turbidity standard lens on the extent of lens opacification grade and score. Gaps in the course of the entire experiment the rats always maintained a transparent lens; experiment the first 10 days to check and found that the model group rats were all basically surrounding lens vacuoles, accounting for 1/3-2/3 of the cortex area, score at 0.1-1 points, the treatment group of individual rat lens is still transparent, and a few other surrounding vacuolated lens, score at 0.1-0.7 points; to experiment the first 20 days, the model group rats appeared punctate lens opacities or sheet, score at 2-4 points, the treatment group than in rat lens vacuoles increased, there was no obvious point of turbidity sheet, score at 0.7-1 points; to experiment the first 30 days, the model group rats were all nuclear lens opacities, the majority of rats nuclear lens opacities all, score at 4-5 points, while the treatment group, only two appear rat lens nuclear lens opacities, and the remaining rat lens opacification only see point sheet, score at 2-4 points; by the statistical test in each group of lens density 10,20,30 score the first day were significantly different (P <0.05).2 Lens of rats in each group of nitric oxide and nitric oxide synthase content.2.1The conten nitric oxide blank group's lens , content of nitric oxide is 0. 452±0.152; model group is 2.666±0.466; the treatment group is 1.683±0.344. The blank group was significantly less than model group and treatment group, model group than the treatment group, with statistical analysis, each group comparison are significantly different; (P <0.05);2.2 The content of nitric oxide synthase blank group's lens, content of nitric oxide synthase is 0. 016±0.002; model group is 0.037±0.004; the treatment group is 0.027±0.001. Nitric oxide synthase in the model group were higher than those of the treatment group, treatment group was higher than blank control group, each group comparison are significantly different; (P <0.05);3 In each group within the rat lens caspase-3 contentDetected by flow cytometry analysis of apoptotic gene caspase-3 ,in blank lens group , content of caspase-3 is 339. 438±37.937; model group is 697.722±46.453; treatment group is 650.748±53.074. Caspase-3 in blank group's lens is low ,in model group, caspase-3 in the highest levels, the treatment group between the two; each group comparison are significantly different; (P <0.05);Conclusion:1 Nitric oxide involved in the occurrence of cataract and the development process.2 Nitric oxide synthase inhibitor, by inhibiting NOS inhibiting NO generation, nhibits apoptosis of lens epithelial cells, a reduction of the lens with the expression of caspase-3,the effective may delayed the occurrence of cataract and development.3 Nitric oxide synthase inhibitor in delaying cataract development happen and has a good application prospects. |
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