白藜芦醇调节微小RNA-29b表达抑制高糖诱导的大鼠视网膜muller细胞凋亡
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摘要
目的研究白藜芦醇(RSV)防护糖尿病早期视损伤的分子机制。方法通过建立视网膜muller细胞(RMCs)体外高糖模型,采用免疫荧光双标、qRT-PCR、western blot、Annexin V/PI流式细胞技术检测高糖和RSV对微小RNA-29b(miR-29b)、特异蛋白1(SP1)表达及RMCs凋亡的影响。结果高糖培养后,miR-29b和SP1表达明显改变,RMCs凋亡率明显增加,RSV干预可明显抑制高糖诱导的miR-29b和SP1异常表达和RMCs凋亡;通过预先在RMCs中转染miR-29b类似物和miR-29b抑制剂研究发现,RSV抑制凋亡作用受明显影响(P<0.05)。结论 RSV通过调节miR-29b表达发挥抗糖尿病早期视损伤作用。
Objective To investigate the molecular mechanisms of protecting early diabetic retinopathy by resvertrol( RSV).Methods The effects of RSV on expressions of miRNA-29b( miR-29b) and specificity protein 1( SP1) as well as apoptosis of retinal muller cells( RMCs) were investigated through development of rat RMCs hyperglucose model in vito,and mothods of immunofluorescence,qRT-PCR,western blot and Annexin V/PI flow-cytometry.Results High glucose induced down-regulation of miR-29 b and upregulation of SP1 and increased the apoptosis of RMCs.The effects could be reversed by RSV in vitro.Moreover,the anti-apoptosis effect of RSV could be reversed by transfection of miR-29 b analogues to RMCs and miR-29 b inhibitor( P < 0. 05).Conclusion Resveratrol can inhibit diabetic retinopathy in early stage by regulation of miR-29 b expression.
Objective To investigate the molecular mechanisms of protecting early diabetic retinopathy by resvertrol( RSV).Methods The effects of RSV on expressions of miRNA-29b( miR-29b) and specificity protein 1( SP1) as well as apoptosis of retinal muller cells( RMCs) were investigated through development of rat RMCs hyperglucose model in vito,and mothods of immunofluorescence,qRT-PCR,western blot and Annexin V/PI flow-cytometry.Results High glucose induced down-regulation of miR-29 b and upregulation of SP1 and increased the apoptosis of RMCs.The effects could be reversed by RSV in vitro.Moreover,the anti-apoptosis effect of RSV could be reversed by transfection of miR-29 b analogues to RMCs and miR-29 b inhibitor( P < 0. 05).Conclusion Resveratrol can inhibit diabetic retinopathy in early stage by regulation of miR-29 b expression.
引文
[1]Zhou T,Che D,Lan Y,et al.Mesenchymal marker expression is elevated in Müller cells exposed to high glucose and in animal models of diabetic retinopathy[J].Oncotarget,2017,8(3):4582-4594.
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[7]Goutham G,Manikandan R,Beulaja M,et al.A focus on resveratrol and ocular problems,especially cataract:From chemistry to medical uses and clinical relevance[J].Biomed Pharmacother,2017,86:232-241.
[8]Ghadiri Soufi F,Arbabi-Aval E,Rezaei Kanavi M,et al.Anti-inflammatory properties of resveratrol in the retinas of type 2 diabetic rats[J].Clin Exp Pharmacol Physiol,2015,42(1):63-68.
[9]Bola C,Bartlett H,Eperjesi F.Resveratrol and the eye:activity and molecular mechanisms[J].Graefes Arch Clin Exp Ophthalmol,2014,252(5):699-713.
[2]赵帅,尹妮.激光光凝治疗糖尿病视网膜病变后的药物干预[J].实用医院临床杂志,2016,13(2):117-118.
[3]Zeng K,Yang N,Wang D,et al.Resveratrol prevents retinal dysfunction by regulating glutamate transporters,glutamine synthetase expression and activity in diabetic retina[J].Neurochem Res,2016,41(5):1050-1064.
[4]Zeng K,Ming J,Yang N,et al.Taurine prevents high glucose-induced angiopoietin-2/tie-2 system alterations and apoptosis in retinal microvascular pericytes[J].Mol Cell Biochem,2014,396(1-2):239-248.
[5]Zeng K,Xu H,Chen K,et al.Effects of taurine on glutamate uptake and degradation in Müller cells under diabetic conditions via antioxidant mechanism[J].Mol Cell Neurosci,2010,45(2):192-199.
[6]Silva VA,Polesskaya A,Sousa TA,et al.Expression and cellular localization of microRNA-29b and RAX,an activator of the RNA-dependent protein kinase(PKR),in the retina of streptozotocin-induced diabetic rats[J].Mol Vis,2011,17:2228-2240.
[7]Goutham G,Manikandan R,Beulaja M,et al.A focus on resveratrol and ocular problems,especially cataract:From chemistry to medical uses and clinical relevance[J].Biomed Pharmacother,2017,86:232-241.
[8]Ghadiri Soufi F,Arbabi-Aval E,Rezaei Kanavi M,et al.Anti-inflammatory properties of resveratrol in the retinas of type 2 diabetic rats[J].Clin Exp Pharmacol Physiol,2015,42(1):63-68.
[9]Bola C,Bartlett H,Eperjesi F.Resveratrol and the eye:activity and molecular mechanisms[J].Graefes Arch Clin Exp Ophthalmol,2014,252(5):699-713.