The role of sirt1 in the retinal ganglion cells cultured by high glucose
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ORIGINAL PAPER
The role of sirt1 in the retinal ganglion cells cultured by high glucose Yue Zhou . Lian-jun Cai . Li-hui Xu . Yu Guo . Nuo Chen . Qing Yao
Received: 18 February 2020 / Accepted: 29 October 2020 Ó Springer Nature B.V. 2020
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-j B, caspase-3 was detected by Western blot.
Yue Zhou and Lian-jun Cai: These authors contributed equally to this work Y. Zhou L. Cai L. Xu Y. Guo Q. Yao (&) Department of Biochemistry and Molecular Biology, School of Basic Medical Science, Ningxia Medical University, Yinchuan, Ningxia, China e-mail: [email protected] Y. Zhou Department of Biochemistry and Molecular Biology, School of Basic Medical Science, Peking University, Beijing, China N. Chen Department of Clinical Medicine, Xiangya School of Medicine, Central South University, Changsha, Hunan, China
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-j 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-jB 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. Keywords Sirt1 Diabetic retinopathy Retinal ganglion cells
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Int Ophthalmol
Introduction
Materials and Methods
Diabetic retinopathy (DR) is the visual impairment disease caused by hyperglycemia [1]. It is one of the most common and serious complications of diabetic mellitus (DM) [2]. Meanwhile, it is a
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