| Background:Fungal keratitis is a serious blinding eye disease caused by fungal infections,which often occurs after plant trauma.If the disease is not treated in a timely and effective manner at the early stage of the infection,it will lead to serious complications,hypha growth into the eye can lead to corneal perforation and fungal endophthalmitis,and 15-27%of patients with fungal keratitis eventually receive corneal transplantation.Removal of ocular contents is an important factor for blindness caused by fungal keratitisRapamycin is a new macrolide immunosuppressant and widely used autophagy-specific inducer,which can promote autophagy by inhibiting mammalian rapamycin target protein(mTOR)expression.Studies have shown that the fusion of mature autophagosomes with lysosomes or vacuoles can promote the degradation of Acanthamoeba trophozoite and exert its anti-Acanthamoeba effect.Cellular autophagy is a self-stable state coexisting in eukaryotes,and plays an important role in the process of cell development,self-protection and growth.Under the action of the external environment,the autophagy pathway is activated,thereby removing the abnormal substances in the cell and maintaining the normal physiological function of the cell.In this process,the autophagy marker microtubule-associated protein light chain 3(LC-3 ?)can be attached to the autophagosome membrane,thereby acting as a structural protein of the autophagosome.Therefore,by detecting the expression of LC-3? in the experiment to reflect the degree of autophagy.Recent studies have shown that autophagy can effectively inhibit the occurrence of fibrotic diseases and reduce the degree of scarring.In this study we explored the effect of the mTOR pathway inhibitor rapamycin on corneal fibrosis of fungal keratitis.This is not only important for preventing and treating blindness,but also helps to understand the mechanism of cellular autophagy in fungal infections,and provides a new program for the drug treatment of fungal keratitis..ObjectTo investigate the effects of mTOR pathway inhibitor rapamycin on corneal fibrosis of fungal keratitis in miceMethods:96 healthy C57BL/6J male mice were selected and randomly divided into a rapamycin group and a control group.The mice were injected intraperitoneally at a concentration of 6.0 mg.kg-1 the day before the rapamycin model of fungal keratitis was created.After pretreatment,5 ?l was injected subconjunctivally at a concentration of 0.2 g·L-1 once a day for 3 days.The control group was controlled with PBS solution.The basic conditions and clinical scores of the cornea were observed under the slit lamp microscope at 1d,2d,3d,4d,5d,6d,7d,9d,and 14d after the model was made.The percentage of corneal area occupied by corneal lesions after infection.The expression levels of LC-3?,?-SMA and TGF-?1 in the cornea were detected by Western Blot and real-time fluorescence quantitative PCR 3d,6d,9d and 14d after modelingResults:The clinical scores of the rapamycin group were lower than those of the control group at 1d,2d,3d,4d,5d,6d and 14d after model.And The difference was statistically significant(all P<0.05).The perforation rate was 47.72%in the control group and 27.78%in the rapamycin group.The difference was statistically significant(P=0.0381).The proportion of lesion area in the rapamycin group was significantly smaller than that in the model control group.The difference was statistically significant(P<0.05).After the injection of rapamycin,the expression of LC-3?increased,and the difference was statistically significant at 6d and 9d,The expression of fibrosis-related factors decreased,and the difference of ?-SMA at 6d and 9d was statistically significant(all P<0.05),and the difference of TGF-?1 at 6d and 14d was Statistically significant(all P<0.05).ConclusionRapamycin can activate the production of autophagy,thereby reducing the perforation rate and the expression of fungal keratitis-fibrosis related factors,and reducing the degree of fibrosis left by fungal keratitis... |
PDF Full Text Request