Abstract
Objective
To observe the effect of sirt1 on retinal ganglion cells (RGC) with high glucose culture and to explore the role of sirt1 in the development of diabetic retinopathy. Method RGC was infected by sirt1 lentivirus overexpression vector pLV5-sirt1 and interference vector pLV3-si-sirt1. The normal control group and control virus vector group were set up at the same time. After 48 h of infection, the viability of RGC was detected by CCK8 kit, the apoptosis rate was detected by FCM analysis, and the protein expression of p53, FOXO3a, NF-κ B, caspase-3 was detected by Western blot.
Results
After RGC were infected with lentivirus, the cell viability of lentivirus overexpression vector pLV5-sirt1 was significantly higher than that of the high glucose group and the sirt1 overexpression control group, while the cell viability of interference vector pLV3-si-sirt1 was significantly lower than that of the high glucose group and the sirt1 interference control group (P < 0.05). At the same time, the apoptosis rate of RGC cells infected by lentivirus overexpression vector pLV5-sirt1 was lower than that of the high glucose group and the control virus vector group, while the apoptosis rate of the interference vector pLV3-si-sirt1 cells was significantly higher than that of the high glucose group and the control virus vector group (P < 0.05). The results of Western blotting showed that the expression of p53, FOXO3a, NF-κ B and caspase-3 in RGC cells decreased significantly after infection with pLV5-sirt1 compared with the high glucose group and the control virus vector group, while the expression of p53, FOXO3a, NF-κB and caspase-3 in RGC cells increased significantly after infection with pLV3-si-sirt1 (P < 0.05).
Conclusion
Sirt1 can inhibit the apoptosis of RGCs through regulating the expression of some apoptotic cytokinessome, and it can be used as a candidate gene for the biotherapy of retinal diseases.
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We acknowledge support from Natural Science Foundation of China (82060819) and Ningxia Key Research and Development Project (2020BEG03033), Innovation and entrepreneurship training program of Ningxia Medical University.
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Yue Zhou and Lianjun Cai performed laboratory testing and edited and checked the accuracy of the manuscript. Lihui Xu and Yu Guo performed laboratory testing. Nuo Chen contributed to the sample collection and data analysis. Qing Yao was responsible for study design and recruitment, contributed to the sample collection and data analysis, and reviewed and edited the manuscript. Qing Yao is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.
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Qing Yao is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.
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Yue Zhou and Lian-jun Cai: These authors contributed equally to this work
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Zhou, Y., Cai, Lj., Xu, Lh. et al. The role of sirt1 in the retinal ganglion cells cultured by high glucose. Int Ophthalmol 41, 845–852 (2021). https://doi.org/10.1007/s10792-020-01638-4
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DOI: https://doi.org/10.1007/s10792-020-01638-4