Effects of etanercept on the apoptosis of ganglion cells and expression of Fas, TNF-α, caspase-8 in the retina of diabetic rats
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摘要
AIM: To evaluate the effects of etanercept on the expression of Fas, tumor necrosis factor-alpha(TNF-α) and caspase-8 in the early stage of the apoptotic pathway in diabetic rats, and to explore the therapeutic effect of etanercept on diabetic retinopathy.METHODS: A total of 60 Sprague-Dawley(SD) rats were randomly and evenly divided into 3 groups with 20 rats each, including control group, and diabetic groups with or without treatment. Streptozotocin(STZ)-induced diabetic rats were established for diabetic groups. Blood glucose and body weight were measured weekly. All the rats were sacrificed at the 12 wk after treatment. The expressions of Fas, TNF-α and caspase-8 in rat retina were quantitatively detected by PCR and Western blot. The leakage of Evan blue was adopted to measure the retinal vascular leakage quantitatively, and to compare it among different groups. TUNEL method was used to compare the amount of apoptotic bodies quantitatively in rat retina ganglion cells under electron microscope.RESULTS: The expressions of Fas, TNF-α and caspase-8 in each group were compared via PCR and Western blot, in which the diabetic group with treatment was lower than those without treatment(P<0.01), but all the diabetic groups were higher than the control group(P<0.01). Evans blue leakage in the diabetic treatment group was lower than those without treatment(P<0.01), but those in the control group was the lowest compared with the other two groups(P<0.01). TUNEL method showed that the apoptoticbodies of retina in the diabetic treatment group was lower than those without treatment(P<0.01), while those in the control group was the lowest compared with the other two groups(P<0.01). CONCLUSION: Etanercept can effectively reduce the expression of Fas, TNF-α and caspase-8, as well as the retinal leakage and retinal cell apoptosis in diabetic rats.
AIM: To evaluate the effects of etanercept on the expression of Fas, tumor necrosis factor-alpha(TNF-α) and caspase-8 in the early stage of the apoptotic pathway in diabetic rats, and to explore the therapeutic effect of etanercept on diabetic retinopathy.METHODS: A total of 60 Sprague-Dawley(SD) rats were randomly and evenly divided into 3 groups with 20 rats each, including control group, and diabetic groups with or without treatment. Streptozotocin(STZ)-induced diabetic rats were established for diabetic groups. Blood glucose and body weight were measured weekly. All the rats were sacrificed at the 12 wk after treatment. The expressions of Fas, TNF-α and caspase-8 in rat retina were quantitatively detected by PCR and Western blot. The leakage of Evan blue was adopted to measure the retinal vascular leakage quantitatively, and to compare it among different groups. TUNEL method was used to compare the amount of apoptotic bodies quantitatively in rat retina ganglion cells under electron microscope.RESULTS: The expressions of Fas, TNF-α and caspase-8 in each group were compared via PCR and Western blot, in which the diabetic group with treatment was lower than those without treatment(P<0.01), but all the diabetic groups were higher than the control group(P<0.01). Evans blue leakage in the diabetic treatment group was lower than those without treatment(P<0.01), but those in the control group was the lowest compared with the other two groups(P<0.01). TUNEL method showed that the apoptoticbodies of retina in the diabetic treatment group was lower than those without treatment(P<0.01), while those in the control group was the lowest compared with the other two groups(P<0.01). CONCLUSION: Etanercept can effectively reduce the expression of Fas, TNF-α and caspase-8, as well as the retinal leakage and retinal cell apoptosis in diabetic rats.
AIM: To evaluate the effects of etanercept on the expression of Fas, tumor necrosis factor-alpha(TNF-α) and caspase-8 in the early stage of the apoptotic pathway in diabetic rats, and to explore the therapeutic effect of etanercept on diabetic retinopathy.METHODS: A total of 60 Sprague-Dawley(SD) rats were randomly and evenly divided into 3 groups with 20 rats each, including control group, and diabetic groups with or without treatment. Streptozotocin(STZ)-induced diabetic rats were established for diabetic groups. Blood glucose and body weight were measured weekly. All the rats were sacrificed at the 12 wk after treatment. The expressions of Fas, TNF-α and caspase-8 in rat retina were quantitatively detected by PCR and Western blot. The leakage of Evan blue was adopted to measure the retinal vascular leakage quantitatively, and to compare it among different groups. TUNEL method was used to compare the amount of apoptotic bodies quantitatively in rat retina ganglion cells under electron microscope.RESULTS: The expressions of Fas, TNF-α and caspase-8 in each group were compared via PCR and Western blot, in which the diabetic group with treatment was lower than those without treatment(P<0.01), but all the diabetic groups were higher than the control group(P<0.01). Evans blue leakage in the diabetic treatment group was lower than those without treatment(P<0.01), but those in the control group was the lowest compared with the other two groups(P<0.01). TUNEL method showed that the apoptoticbodies of retina in the diabetic treatment group was lower than those without treatment(P<0.01), while those in the control group was the lowest compared with the other two groups(P<0.01). CONCLUSION: Etanercept can effectively reduce the expression of Fas, TNF-α and caspase-8, as well as the retinal leakage and retinal cell apoptosis in diabetic rats.
引文
1 Wiwanitkit V.IL-2 and IFN-gamma and diabetic retinopathy.Graefes Arch Clin Exp Ophthalmol 2011;249(3):459.
2 Ferrari D,Speciale A,Cristani M,Fratantonio D,Molonia MS,Ranaldi G,Saija A,Cimino F.Cyanidin-3-O-glucoside inhibits NF-κB signalling in intestinal epithelial cells exposed to TNF-αand exerts protective effects via Nrf2 pathway activation.Toxicol Lett 2016;264:51-58.
3 Stitt AW,Curtis TM,Chen M,Medina RJ,McKay GJ,Jenkins A,Gardiner TA,Lyons TJ,Hammes HP,SimóR,Lois N.The progress in understanding and treatment of diabetic retinopathy.Prog Retin Eye Res2016;51:156-186.
4 Lee YJ,Jung SH,Kim SH,Kim MS,Lee S,Hwang J,Kim SY,Kim YM,Ha KS.Essential role of transglutaminase 2 in vascular endothelial growth factor-induced vascular leakage in the retina of diabetic mice.Diabetes 2016;65(8):2414-2428.
5 Akash MSH,Rehman K,Liaqat A.Tumor necrosis factor-alpha:role in development of insulin resistance and pathogenesis of type 2 diabetes mellitus.J Cell Biochem 2018;119(1):105-110.
6 Tavares Ferreira J,Alves M,Dias-Santos A,Costa L,Santos BO,Cunha JP,Papoila AL,Abeg?o Pinto L.Retinal neurodegeneration in diabetic patients without diabetic retinopathy.Invest Ophthalmol Vis Sci2016;57(14):6455.
7 Zhang Y,Bi HS,Pan HW,Pan XM,Feng YQ,Liu L,Wang SM.Research of serum total and high-molecular-weight adiponectin levels in patients with type 2 diabetic retinopathy.Guoji Yanke Zazhi(Int Eye Sci)2016;16(3):403-408.
8 Yang QH,Zhang Y,Zhang XM,Li XR.Prevalence of diabetic retinopathy,proliferative diabetic retinopathy and non-proliferative diabetic retinopathy in Asian T2DM patients:a systematic review and Meta-analysis.Int J Ophthalmol 2019;12(2):302-311
9 Heitmar R,Vekria P,Cubbidge RP.Regulation of oxygen saturation in retinal blood vessels in response to dynamic exercise.Acta Ophthalmol2018;96(3):e298-e303.
10 Silverman SM,Wong WT.Microglia in the retina:roles in development,maturity,and disease.Annu Rev Vis Sci 2018;4:45-77.
11 Li Q,Li Z,Zhang SW.Regulation of VEGF of puerarin on STZdiabetic rats retinal and the promoting apoptosis study of puerarin on STZ diabetic rats retinal.Journal of Clinical and Experimental Medicine2017;17:1679-1682.
12 Phyu HE,Irwin JC,Vella RK,Fenning AS.Resveratrol shows neuronal and vascular-protective effects in older,obese,streptozotocininduced diabetic rats.Br J Nutr 2016;115(11):1911-1918.
13 Ye FX,Kaneko H,Hayashi Y,Takayama K,Hwang SJ,Nishizawa Y,Kimoto R,Nagasaka Y,Tsunekawa T,Matsuura T,Yasukawa T,Kondo T,Terasaki H.Malondialdehyde induces autophagy dysfunction and VEGF secretion in the retinal pigment epithelium in age-related macular degeneration.Free Radic Biol Med 2016;94:121-134.
14 Zhu K,Zhang ML,Liu ST,Li XY,Zhong SM,Li F,Xu GZ,Wang ZF,Miao YY.Ghrelin attenuates retinal neuronal autophagy and apoptosis in an experimental rat glaucoma model.Invest Ophthalmol Vis Sci2017;58(14):6113-6122.
15 Nita M,Grzybowski A.The role of the reactive oxygen species and oxidative stress in the pathomechanism of the age-related ocular diseases and other pathologies of the anterior and posterior eye segments in adults.Oxid Med Cell Longev 2016;2016:3164734.
16 Yin XM.Bid,a critical mediator for apoptosis induced by the activation of Fas/TNF-R1 death receptors in hepatocytes.J Mol Med2000;78(4):203-211.
17 Sartorius U,Schmitz I,Krammer PH.Molecular mechanisms of deathreceptor-mediated apoptosis.Chembiochem 2001;2(1):20-29.
18 Hu YY,Li QM.Research research progress in the pathogenesis of diabetic peripheral neuropathy.Medical Recapitulate 2016;20:598-602.
19 Behl Y,Krothapalli P,Desta T,DiPiazza A,Roy S,Graves DT.Diabetes-enhanced tumor necrosis factor-alpha production promotes apoptosis and the loss of retinal microvascular cells in type 1 and type 2models of diabetic retinopathy.Am J Pathol 2008;172(5):1411-1418.
20 Murdaca G,Negrini S,Magnani O,Penza E,Pellecchio M,Gulli R,Mandich P,Puppo F.Update upon efficacy and safety of etanercept for the treatment of spondyloarthritis and juvenile idiopathic arthritis.Mod Rheumatol 2018;28(3):417-431.
2 Ferrari D,Speciale A,Cristani M,Fratantonio D,Molonia MS,Ranaldi G,Saija A,Cimino F.Cyanidin-3-O-glucoside inhibits NF-κB signalling in intestinal epithelial cells exposed to TNF-αand exerts protective effects via Nrf2 pathway activation.Toxicol Lett 2016;264:51-58.
3 Stitt AW,Curtis TM,Chen M,Medina RJ,McKay GJ,Jenkins A,Gardiner TA,Lyons TJ,Hammes HP,SimóR,Lois N.The progress in understanding and treatment of diabetic retinopathy.Prog Retin Eye Res2016;51:156-186.
4 Lee YJ,Jung SH,Kim SH,Kim MS,Lee S,Hwang J,Kim SY,Kim YM,Ha KS.Essential role of transglutaminase 2 in vascular endothelial growth factor-induced vascular leakage in the retina of diabetic mice.Diabetes 2016;65(8):2414-2428.
5 Akash MSH,Rehman K,Liaqat A.Tumor necrosis factor-alpha:role in development of insulin resistance and pathogenesis of type 2 diabetes mellitus.J Cell Biochem 2018;119(1):105-110.
6 Tavares Ferreira J,Alves M,Dias-Santos A,Costa L,Santos BO,Cunha JP,Papoila AL,Abeg?o Pinto L.Retinal neurodegeneration in diabetic patients without diabetic retinopathy.Invest Ophthalmol Vis Sci2016;57(14):6455.
7 Zhang Y,Bi HS,Pan HW,Pan XM,Feng YQ,Liu L,Wang SM.Research of serum total and high-molecular-weight adiponectin levels in patients with type 2 diabetic retinopathy.Guoji Yanke Zazhi(Int Eye Sci)2016;16(3):403-408.
8 Yang QH,Zhang Y,Zhang XM,Li XR.Prevalence of diabetic retinopathy,proliferative diabetic retinopathy and non-proliferative diabetic retinopathy in Asian T2DM patients:a systematic review and Meta-analysis.Int J Ophthalmol 2019;12(2):302-311
9 Heitmar R,Vekria P,Cubbidge RP.Regulation of oxygen saturation in retinal blood vessels in response to dynamic exercise.Acta Ophthalmol2018;96(3):e298-e303.
10 Silverman SM,Wong WT.Microglia in the retina:roles in development,maturity,and disease.Annu Rev Vis Sci 2018;4:45-77.
11 Li Q,Li Z,Zhang SW.Regulation of VEGF of puerarin on STZdiabetic rats retinal and the promoting apoptosis study of puerarin on STZ diabetic rats retinal.Journal of Clinical and Experimental Medicine2017;17:1679-1682.
12 Phyu HE,Irwin JC,Vella RK,Fenning AS.Resveratrol shows neuronal and vascular-protective effects in older,obese,streptozotocininduced diabetic rats.Br J Nutr 2016;115(11):1911-1918.
13 Ye FX,Kaneko H,Hayashi Y,Takayama K,Hwang SJ,Nishizawa Y,Kimoto R,Nagasaka Y,Tsunekawa T,Matsuura T,Yasukawa T,Kondo T,Terasaki H.Malondialdehyde induces autophagy dysfunction and VEGF secretion in the retinal pigment epithelium in age-related macular degeneration.Free Radic Biol Med 2016;94:121-134.
14 Zhu K,Zhang ML,Liu ST,Li XY,Zhong SM,Li F,Xu GZ,Wang ZF,Miao YY.Ghrelin attenuates retinal neuronal autophagy and apoptosis in an experimental rat glaucoma model.Invest Ophthalmol Vis Sci2017;58(14):6113-6122.
15 Nita M,Grzybowski A.The role of the reactive oxygen species and oxidative stress in the pathomechanism of the age-related ocular diseases and other pathologies of the anterior and posterior eye segments in adults.Oxid Med Cell Longev 2016;2016:3164734.
16 Yin XM.Bid,a critical mediator for apoptosis induced by the activation of Fas/TNF-R1 death receptors in hepatocytes.J Mol Med2000;78(4):203-211.
17 Sartorius U,Schmitz I,Krammer PH.Molecular mechanisms of deathreceptor-mediated apoptosis.Chembiochem 2001;2(1):20-29.
18 Hu YY,Li QM.Research research progress in the pathogenesis of diabetic peripheral neuropathy.Medical Recapitulate 2016;20:598-602.
19 Behl Y,Krothapalli P,Desta T,DiPiazza A,Roy S,Graves DT.Diabetes-enhanced tumor necrosis factor-alpha production promotes apoptosis and the loss of retinal microvascular cells in type 1 and type 2models of diabetic retinopathy.Am J Pathol 2008;172(5):1411-1418.
20 Murdaca G,Negrini S,Magnani O,Penza E,Pellecchio M,Gulli R,Mandich P,Puppo F.Update upon efficacy and safety of etanercept for the treatment of spondyloarthritis and juvenile idiopathic arthritis.Mod Rheumatol 2018;28(3):417-431.