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Part1Ovariectomized Rat Model Making and Ocular Surface EvaluationObjective To make ovariectomized rat model and to observe the influence of ocular surface in rat when the level of sex hormones change.Methods Twenty female healthy SD rats were divided into the contol group(n=10) and the experiment group(n=10) randomly. Ovariectomy was performed in the experiment group,while in the control group,the sham operation was done. At the3rd month,the concentration of estrogen and androgen were measured by radiation immunoassay detection method. Schirmer I test and corneal fluorescence staining examination were done in all rats before operation and at1,2, and3months after ovariectomized.Result Compared with the control group, the concentration of estrogen in the experiment group was decreased,showing statistically difference (t=-35.37,P<0.05). The concentration of androgen in the experiment group was decreased,showing statistically difference (t=-12.13,P<0.05).Compared with the control group, the experiment group at the1m,2m and3m time points didn’t show difference either in Schirmer I test or in corneal fluorescence staining examination (P>0.05).Conclusion The level of estrogen and androgen in the experiment group dropped after three months of surgey while no obvious clinical damage signs of ocular surface were found. Part2The Expression of Inflammatory Factors in Conjunctiva in Ovariectomized RatObjective To ovserve the expression of inflammatory factors:IL-17A, IL-1β, IL-6and TNF-α in conjunctiva in ovariectomized rat.Methods Twenty female healthy SD rats were divided into the contol group(n=10) and the experiment group(n=10) randomly. Ovariectomy was performed in the experiment group,while in the control group,the sham operation was done. The bulbar conjunctivas under the upper palpebral fissure were obtained at the3rd month after ovariectomized.Conjunctival epithelial cell morphology was examined by HE staining.The exprssions of IL-17A, IL-1β, IL-6and TNF-α were analyzed by immunohistochemical method and Western blotting analysis.Result The layer numbers of the conjunctival epithelial cell increasing, cell arrangement disorder and angle trend were observed in the experiment group by HE staining.By immunohistochemical method,the expression of inflammatory factors in conjunctivas in the experiment group were found all higher than those in the control group (A value:IL-17A:t=8.22,P<0.01、IL-1β:t=16.43,P<0.01、IL-6:t=13.44,P<0.01、 TNF-α:t=16.26,P<0.01).By Western blot method,the expression levels of inflammatory factors in conjunctivas in the experiment group were found all higher than those in the control group (A value:IL-17A:t=19.41,P<0.01、IL-1β:t=12.63,P<0.01、 IL-6:t=17.17,P<0.01、TNF-α:t=15.19,P<0.01).Conclusion In ovariectomized rat, conjunctival histologic change appeared and the expression levels of inflammatory factors,such as IL-17A, IL-1β, IL-6and TNF-α were increased. Part3The Expression of Inflammatory Factors in Lachrymal Gland in Ovariectomized RatObjective To ovserve the expression of inflammatory factors:IL-17A, IL-1β, IL-6and TNF-α in lachrymal gland in ovariectomized rat.Methods Twenty female healthy SD rats were divided into the contol group(n=10) and the experiment group(n=10) randomly. Ovariectomy was performed in the experiment group,while in the control group,the sham operation was done. Lachrymal glands were obtained at the3rd month after ovariectomized. Acinar cells of the lacrimal glands was examined by HE staining.The exprssions of IL-17A, IL-1β, IL-6and TNF-α were analyzed by immunohistochemical method and Western blotting analysis.Result By HE staining observation, the enzyme original particle in lachrymal cell significantly reduced while acinar atrophy and cell arrangement disorder were observed in the experiment group. By immunohistochemical method,the expression of inflammatory factors in lachrymal gland in the experiment group were found all higher than those in the control group (A value:IL-17A:t=11.71,P<0.01、IL-1β:t=15.02,P<0.01IL-6:t=25.59,P<0.01、TNF-α:t=14.44,P<0.01). By Western blot method,the expression levels of inflammatory factors in lachrymal gland in the experiment group were found all higher than those in the control group (A value:IL-17A:t=25.82,P<0.01、 IL-1β:t=25.87,P<0.01、IL-6:t=22.95,P<0.01、TNF-α:t=20.90,P<0.01).Conclusion In ovariectomized rat, lacrimal histologic change appeared and the expression levels of inflammatory factors,such as IL-17A, IL-1β, IL-6and TNF-α were increased.
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