1型糖尿病大鼠不同时期腰髓线粒体动力学变化的研究
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摘要
目的观察T1DM大鼠不同时期腰髓中线粒体形态、数量及动力学变化。方法 50只雄性SD大鼠腹腔注射STZ建立T1DM大鼠模型,成模后随机分为糖尿病4周组(DM 4W)和糖尿病12周组(DM 12W)。另取24只雄性SD大鼠随机分为正常4周组(NC 4W,n=12)和正常12周组(NC 12W,n=12)。观察线粒体形态、数量并检测线粒体动力蛋白1(Drp1)、线粒体融合蛋白1(Mfn1)和线粒体标志物(VDAC1)的表达。结果随着时间增加,DM 4W组和DM 12W组体重减少[(259. 08±12. 41)vs(243. 92±9. 84)g,(260. 17±10. 65)vs(194. 17±8. 17)g,P<0. 01]。与NC 4W比较,DM 4W组线粒体形态破坏,数量增加(P<0. 01),Drp1和VDAC1表达升高(P<0. 01),Mfn1表达降低(P<0. 01)。与NC 12W组比较,DM 12W组线粒体形态破坏,数量减少(P<0. 01),Drp1、Mfn1和VDAC1表达降低(P<0. 01)。结论随着病程延长,线粒体形态严重损害,数量由多变少和动力学失衡,这些改变可能成为影响神经系统功能正常发挥的因素之一。
Objective To observe the changes of mitochondria morphology,quantity and dynamics in the lumbar spinal cord of type 1 diabetic(T1DM)rats at different stages.Methods Male SD rats were injected intraperitoneally with streptozotocin to establish T1DM model. After modeling,they were randomly divided into the diabetic 4-week(DM 4W)group and the diabetic 12-week(DM 12 W)group. A normal control (NC) group was also provided. Morphology and quantity of mitochondria were observed and the expressions of mitochondrial motor protein 1(Drp1),mitochondrial fusion protein 1(Mfn1)and mitochondrial marker(VDAC1)were examined.Results As time goes on,weight loss increases. Compared with base-line,the body weight of DM 4W group[(259. 08±12. 41)vs(243. 92±9. 84)g,P<0. 01]and DM 12W group[(260. 17±10. 65)vs(194. 17±8. 17)g,P<0. 01]decreased. Compared with NC 4W group,the morphological damage of mitochondria in DM 4W group was obvious,the number increased(P<0. 01),the expression of Drp1 and VDAC1 increased(P<0. 01),and the expression of Mfn1 decreased(P<0. 01).Compared with NC 12W group,the morphological damage of mitochondria in DM 12W group was more obvious,the number decreased(P<0. 01),and the expression of Drp1,Mfn1 and VDAC1 decreased(P<0. 01).Conclusion As the course of T1DM prolongs,the mitochondrial morphology is gradually aggravated,the number is reduced and the kinetics is out of balance. These changes may be factors in the pathogenesis of the nervous system.
Objective To observe the changes of mitochondria morphology,quantity and dynamics in the lumbar spinal cord of type 1 diabetic(T1DM)rats at different stages.Methods Male SD rats were injected intraperitoneally with streptozotocin to establish T1DM model. After modeling,they were randomly divided into the diabetic 4-week(DM 4W)group and the diabetic 12-week(DM 12 W)group. A normal control (NC) group was also provided. Morphology and quantity of mitochondria were observed and the expressions of mitochondrial motor protein 1(Drp1),mitochondrial fusion protein 1(Mfn1)and mitochondrial marker(VDAC1)were examined.Results As time goes on,weight loss increases. Compared with base-line,the body weight of DM 4W group[(259. 08±12. 41)vs(243. 92±9. 84)g,P<0. 01]and DM 12W group[(260. 17±10. 65)vs(194. 17±8. 17)g,P<0. 01]decreased. Compared with NC 4W group,the morphological damage of mitochondria in DM 4W group was obvious,the number increased(P<0. 01),the expression of Drp1 and VDAC1 increased(P<0. 01),and the expression of Mfn1 decreased(P<0. 01).Compared with NC 12W group,the morphological damage of mitochondria in DM 12W group was more obvious,the number decreased(P<0. 01),and the expression of Drp1,Mfn1 and VDAC1 decreased(P<0. 01).Conclusion As the course of T1DM prolongs,the mitochondrial morphology is gradually aggravated,the number is reduced and the kinetics is out of balance. These changes may be factors in the pathogenesis of the nervous system.
引文
[1]Behl T,Kotwani A.Proposed mechanisms of terminalia catappa in hyperglycaemia and associated diabetic complications.J Pharm Pharmacol,2017,69:123-134.
[2]Sung JY,Tani J,Chang TS,et al.Uncovering sensory axonal dysfunction in asymptomatic type 2 diabetic neuropathy.PLoSOne,2017,12:e0171223.
[3]Dahl R,Larsen S,Dohlmann TL,et al.Three-dimensional reconstruction of the human skeletal muscle mitochondrial network as a tool to assess mitochondrial content and structural organization.Acta Physiol(Oxf),2015,213:145-155.
[4]Hung HL,Cheng SY,Cheung YT,et al.A reciprocal relationship between reactive oxygen species and mitochondrial dynamics in neurodegeneration.Redox Biol,2018,14:7-19.
[5]Ding M,Dong Q,Liu Z.et al.Inhibition of dynamin-related protein 1 protects against myocardial ischemia-reperfusion injury in diabetic mice.Cardiovasc Diabetol,2017,16:19.
[6]Guilford BL,Ryals JM,Lezi E,et al.Dorsal root ganglia mito-chondrial biochemical changes in nondiabetic and streptozotocin-induced diabetic mice fed with a standard or high-fat diet.J Neu-rol Neurosci,2017,8:180.
[7]Sifuentes-Franco S,Pacheco-Moisés FP,Rodríguez-Carrizalez AD,et al.The role of oxidative stress,mitochondrial function,and autophagy in diabetic polyneuropathy.J Diabetes Res,2017,2017:1673081.
[8]Rüb C,Wilkening A,Voos W.Mitochondrial quality control by the Pink1/Parkin system.Cell Tissue Res,2017,367:111-123.
[9]Twig G,Elorza A,Molina AJ,et al.Fission and selective fusion govern mitochondrial segregation and elimination by autophagy.EMBO J,2008,27:433-446.
[10]Youle RJ,van der Bliek AM.Mitochondrial fission,fusion,and stress.Science,2012,337:1062-1065.
[11]Twig G,Graf SA,Wikstrom JD,et al.Tagging and tracking indi-vidual networks within a complex mitochondrialweb with photoac-tivatable GFP.Am J Physiol Cell Physiol,2006,291:C176-C184.
[12]Karbowski M,Arnoult D,Chen H,et al.Quantitation of mito-chondrial dynamics by photolabeling of individual organelles shows that mitochondrial fusion is blocked during the Bax activa-tion phase of apoptosis.J Cell Biol,2004,164:493-499.
[13]Lovy A,Molina AJ,Cerqueira FM,et al.A faster,high resolu-tion,mtPA-GFP-based mitochondrial fusion assay acquiring ki-netic data of multiple cells in parallel using confocal microscopy.J Vis Exp,2012,65:e3991.
[14]Meyer JN,Leuthner TC,Luz AL.Mitochondrial fusion,fis-sion,and mitochondrial toxicity.Toxicology,2017,391:42-53.
[15]Cipolat S,Martins de Brito O,Dal Zilio B,et al.OPA1 re-quires mitofusin 1 to promote mitochondrial fusion.Proc Natl Acad Sci USA,2004,101:15927-15932.
[16]Taguchi N,Ishihara N,Jofuku A,et al.Mitotic phosphorylation of dynamin-related GTPase Drp1 participates in mitochondrial fis-sion.J Biol Chem,2007,282:11521-11529.
[17]Lackner LL.Determining the shape and cellular distribution of mitochondria:the integration of multiple activities.Curr Opin Cell Biol,2013,25:471-476.
[18]Liesa M,Palacín M,Zorzano A.Mitochondrial dynamics in mammalian health and disease.Physiol Rev,2009,89:799-845.
[19]Ingerman E,Perkins EM,Marino M,et al.Dnm1 forms spirals that are structurally tailored to fit mitochondria.J Cell Biol,2005,170:1021-1027.
[20]Vanstone JR,Smith AM,McBride S,et al.DNM1L-related mitochondrial fission defect presenting as refractoryepilepsy.Eur JHum Genet,2016,24:1084-1088.
[21]Rehni AK,Nautiyal N,Perez-Pinzon MA,et al.Hyperglycemia/hypoglycemiainduced mitochondrial dysfunction and cerebral isch-emic damage in diabetics.Metab Brain Dis,2015,30:437-447.
[22]Weng TY,Tsai SA,Su TP.Roles of sigma-1 receptors on mitochondrial functions relevant to neurodegenerative diseases.JBiomed Sci,2017,24:74.
[23]Kou ZZ,Li CY,Hu JC,et al.Alterations in the neural circuits from peripheral afferents to the spinal cord:possible implications for diabetic polyneuropathy in streptozotocin-induced type 1 dia-betic rats.Front Neural Circuits,2014,8:174-175.
[24]Waseem M,Kaushik P,Tabassum H,et al.Role of mitochondri-al mechanism in chemotherapy-induced peripheral neuropathy.Curr Drug Metab,2018,19:47-54.
[25]Russell JW,Zilliox LA.Diabetic neuropathies.Continuum(min-neap minn),2014,20:1226-1240.
[26]Fujimaki S,Kuwabara T,Diabetes-induced dysfunction of mito-chondria and stem cells in skeletal muscle and the nervous sys-tem.Int J Mol Sci,2017,18:2147.
[27]Sifuentes-Franco S,Padilla-Tejeda DE.Oxidative stress,apopto-sis,and mitochondrial function in diabetic nephropathy.Int J En-docrinol,2018,2018:1875870.
[2]Sung JY,Tani J,Chang TS,et al.Uncovering sensory axonal dysfunction in asymptomatic type 2 diabetic neuropathy.PLoSOne,2017,12:e0171223.
[3]Dahl R,Larsen S,Dohlmann TL,et al.Three-dimensional reconstruction of the human skeletal muscle mitochondrial network as a tool to assess mitochondrial content and structural organization.Acta Physiol(Oxf),2015,213:145-155.
[4]Hung HL,Cheng SY,Cheung YT,et al.A reciprocal relationship between reactive oxygen species and mitochondrial dynamics in neurodegeneration.Redox Biol,2018,14:7-19.
[5]Ding M,Dong Q,Liu Z.et al.Inhibition of dynamin-related protein 1 protects against myocardial ischemia-reperfusion injury in diabetic mice.Cardiovasc Diabetol,2017,16:19.
[6]Guilford BL,Ryals JM,Lezi E,et al.Dorsal root ganglia mito-chondrial biochemical changes in nondiabetic and streptozotocin-induced diabetic mice fed with a standard or high-fat diet.J Neu-rol Neurosci,2017,8:180.
[7]Sifuentes-Franco S,Pacheco-Moisés FP,Rodríguez-Carrizalez AD,et al.The role of oxidative stress,mitochondrial function,and autophagy in diabetic polyneuropathy.J Diabetes Res,2017,2017:1673081.
[8]Rüb C,Wilkening A,Voos W.Mitochondrial quality control by the Pink1/Parkin system.Cell Tissue Res,2017,367:111-123.
[9]Twig G,Elorza A,Molina AJ,et al.Fission and selective fusion govern mitochondrial segregation and elimination by autophagy.EMBO J,2008,27:433-446.
[10]Youle RJ,van der Bliek AM.Mitochondrial fission,fusion,and stress.Science,2012,337:1062-1065.
[11]Twig G,Graf SA,Wikstrom JD,et al.Tagging and tracking indi-vidual networks within a complex mitochondrialweb with photoac-tivatable GFP.Am J Physiol Cell Physiol,2006,291:C176-C184.
[12]Karbowski M,Arnoult D,Chen H,et al.Quantitation of mito-chondrial dynamics by photolabeling of individual organelles shows that mitochondrial fusion is blocked during the Bax activa-tion phase of apoptosis.J Cell Biol,2004,164:493-499.
[13]Lovy A,Molina AJ,Cerqueira FM,et al.A faster,high resolu-tion,mtPA-GFP-based mitochondrial fusion assay acquiring ki-netic data of multiple cells in parallel using confocal microscopy.J Vis Exp,2012,65:e3991.
[14]Meyer JN,Leuthner TC,Luz AL.Mitochondrial fusion,fis-sion,and mitochondrial toxicity.Toxicology,2017,391:42-53.
[15]Cipolat S,Martins de Brito O,Dal Zilio B,et al.OPA1 re-quires mitofusin 1 to promote mitochondrial fusion.Proc Natl Acad Sci USA,2004,101:15927-15932.
[16]Taguchi N,Ishihara N,Jofuku A,et al.Mitotic phosphorylation of dynamin-related GTPase Drp1 participates in mitochondrial fis-sion.J Biol Chem,2007,282:11521-11529.
[17]Lackner LL.Determining the shape and cellular distribution of mitochondria:the integration of multiple activities.Curr Opin Cell Biol,2013,25:471-476.
[18]Liesa M,Palacín M,Zorzano A.Mitochondrial dynamics in mammalian health and disease.Physiol Rev,2009,89:799-845.
[19]Ingerman E,Perkins EM,Marino M,et al.Dnm1 forms spirals that are structurally tailored to fit mitochondria.J Cell Biol,2005,170:1021-1027.
[20]Vanstone JR,Smith AM,McBride S,et al.DNM1L-related mitochondrial fission defect presenting as refractoryepilepsy.Eur JHum Genet,2016,24:1084-1088.
[21]Rehni AK,Nautiyal N,Perez-Pinzon MA,et al.Hyperglycemia/hypoglycemiainduced mitochondrial dysfunction and cerebral isch-emic damage in diabetics.Metab Brain Dis,2015,30:437-447.
[22]Weng TY,Tsai SA,Su TP.Roles of sigma-1 receptors on mitochondrial functions relevant to neurodegenerative diseases.JBiomed Sci,2017,24:74.
[23]Kou ZZ,Li CY,Hu JC,et al.Alterations in the neural circuits from peripheral afferents to the spinal cord:possible implications for diabetic polyneuropathy in streptozotocin-induced type 1 dia-betic rats.Front Neural Circuits,2014,8:174-175.
[24]Waseem M,Kaushik P,Tabassum H,et al.Role of mitochondri-al mechanism in chemotherapy-induced peripheral neuropathy.Curr Drug Metab,2018,19:47-54.
[25]Russell JW,Zilliox LA.Diabetic neuropathies.Continuum(min-neap minn),2014,20:1226-1240.
[26]Fujimaki S,Kuwabara T,Diabetes-induced dysfunction of mito-chondria and stem cells in skeletal muscle and the nervous sys-tem.Int J Mol Sci,2017,18:2147.
[27]Sifuentes-Franco S,Padilla-Tejeda DE.Oxidative stress,apopto-sis,and mitochondrial function in diabetic nephropathy.Int J En-docrinol,2018,2018:1875870.