| Objective: To establish a rat model of diabetes,detect the expression of LOX in the retina of diabetes rats,and explore whether LOX is involved in the process of retinal neurodegeneration in diabetes rats.Methods: 1.The model of type II diabetes was established by intraperitoneal injection of STZ in SD rats fed with high fat and high sugar;2.The rats were divided into 4 experimental groups: normal rats(WT),diabetes rats(DM),diabetes rats injected with adenovirus(DM+AAV2-LOX sh RNA1-EGFP),diabetes rats injected with control virus(DM+AAV2-EGFP NC),and the rats were intervened by vitreous injection of AAV2-LOX sh RNA1-EGFP and AAV2-EGFP NC;3.Observe the retinal function of each group by flash ERG;4.The thickness of each layer of retina and the number of RGCs were observed by HE staining of paraffin sections;5.The expression of retinal LOX protein in each group was observed by immunohistochemistry;6.The m RNA and protein expressions of LOX,cleaved Caspase-3,Bax and Bcl-2 were detected by q RT PCR and Western blot.Results: 1.Three days after modeling,the measured fasting blood glucose was more than 16.7mmol/L,and the urine volume and drinking water increased significantly,indicating that the modeling was successful;2.After vitreous injection of adenovirus(DM+AAV2-LOX sh RNA1-EGFP)and control virus(DM+AAV2-EGFP NC)in diabetes rats,the expression of LOX in DM+AAV2-LOX sh RNA1-EGFP group was lower than that in DM group by q RT PCR and Western blot(n=6;*P<0.05).There was no statistical difference between DM+AAV2-EGFP NC group and DM group;3.The results of flash ERG in DM group were lower than those in WT group,and the amplitudes of a wave and b wave in DM + AAV2-LOX sh RNA1-EGFP group were recovered than those in DM group(n=6;*P<0.05).There was no significant difference between DM + r AAV2 EGFP group and DM group;4.Compared with WT group,he staining of eyeball paraffin section in DM group had unclear retinal structure,thinner thickness and less RGCs.The retinal thickness in DM + AAV2-LOX sh RNA1-EGFP group was recovered compared with DM group(n=6;*P< 0.05).There was no significant difference between DM + r AAV2 EGFP group and DM group;5.The expression of LOX protein in DM group was higher than that in WT group,and the expression of LOX protein in DM + AAV2-LOX sh RNA1-EGFP group was lower than that in DM group(n=6;*P<0.05).There was no significant difference between DM + r AAV2 EGFP group and DM group;6.The expressions of LOX m RNA,cleaved caspase-3 m RNA and Bax / Bcl-2 in DM group were higher than those in WT group,and the expressions of LOX m RNA,cleaved caspase-3m RNA and Bax / Bcl-2 in DM + AAV2-LOX sh RNA1-EGFP group were lower than those in DM group(n=6;*P<0.05).There was no significant difference between DM+ AAV2-EGFP NC group and DM group.The expressions of LOX protein,cleaved caspase-3 protein and Bax / Bcl-2 in DM group were higher than those in WT group,and the expressions of LOX protein,cleaved caspase-3 protein and Bax / Bcl-2 in DM+ AAV2-LOX sh RNA1-EGFP group were lower than those in DM group(n=6;*P<0.05).There was no significant difference between DM + r AAV2 EGFP group and DM group.Conclusions:(1)The expression of LOX in the retina of diabetic rats increased.(2)LOX may be involved in diabetic retinal nerve cell apoptosis induced by diabetes.(3)Inhibition of LOX expression may play a protective role in diabetic retinopathy induced by diabetes. |
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