| Aim: The clinical trials results in our group showed safety and effective,most of patients revealed significant improvement in visual function,but there still some patients remain the same poor eyesight.In order to solve this clinical problem,we will complement an animal experiment to determine the optimal injection scheme,which will help the future clinical practice.Methods: 160 mice were selected and divided into experimental groups,negative control groups and blank control groups.The experimental groups 1-3 were respectively injected genetic drug(r AAV2-ND4)according to the following schemes: single vitreous injection in one eye;double vitreous injection in one eye;single vitreous injection in both eyes.The corresponding negative control group was injected with PBS according to the same protocol.Safety indicators: anterior segment;vitreous pressure;retinal HE staining;body weight;content of AAV2 in liver and kidney tissues.Effectiveness indicators: ND4 gene expression,NADH dehydrogenase activity,and green fluorescent protein expression.Safety: The intraocular pressure,the advert event in the anterior segment,and the retinal morphology between the experimental group and the control group were within the normal range;there were no significant difference in body weight changes and content of AAV2 in liver and kidney tissues between the experimental group and the control group.Effectivity: Compared with the normal group,the ND4 gene expression in the experimental groups 1-3 were increased by 67.08,70.04,and 15.23 times,respectively.Compared with the negative control group,the NADH dehydrogenase activity of the experimental groups 1-3 increased by 4.84%,38.91%,and 4.91%,respectively.The expression of green fluorescent protein in experimental group 2 was significantly higher than that either of the other two experimental groups.Conclusion: r AAV2-ND4 is safe in either way of three schemes.When single-eye was double-injected,there existed an optimum gene expression. |
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