人脐带间充质干细胞对1型糖尿病小鼠皮肤组织AGEs/RAGE/NF-κB信号通路影响的研究
|
摘要
目的观察人脐带间充质干细胞(hUCMSCs)对NOD小鼠皮肤组织AGEs/RAGE/NF-κB信号通路的影响,探讨其对于改善糖尿病皮肤组织"隐性损害"的机制。方法 NOD小鼠43只,饲养9周后分为正常对照组(NC组,饲养9周后未发病的NOD小鼠,n=12)和模型组(DM组,饲养后糖尿病发病的NOD小鼠,n=31);DM组再分为干细胞组(MSC,n=11)、胰岛素组(INS,n=10)及空白对照组(BC,n=10)。各组予相应干预,8周后检测随机血糖,并取皮肤组织,光镜下观察结构;ELISA检测AGEs水平;RT-qPCR检测RAGE、NF-κB p65、白介素6(IL-6)、TNF-αmRNA表达水平。结果镜下观察,BC组皮肤组织出现结构紊乱,厚度变薄,细胞肿胀,胶原稀疏等病理改变,MSC组对比前者则有明显改善。与BC组比较,其余3组随机血糖、皮肤AGEs水平及上述4种因子mRNA的表达水平均降低(P<0. 05);与MSC组比较,INS组均增高(P<0. 05)。结论 h UCMSCs可一定程度上抑制AGEs/RAGE/NF-κB信号通路,该作用可能是改善糖尿病皮肤组织"隐性损害"的机制之一。
ObjectiveTo observe the effects of human umbilical cord mesenchymal stem cells(h UCMSCs)on AGEs/RAGE/NF-κB signaling pathway in NOD mice skin tissue,and to explore its effect on improving the"hidden damage"of diabetic skin tissue. mechanism.MethodsA total of 43 NOD mice were divided into normal control group(NC,n=12)and diabetic model group(DM,n=31)after 9 weeks of feeding. The model group was further divided into stem cell treatment group(MSC,n=11),insulin treatment group(INS,n=10)and blank control group(BC,n=10). Random blood glucose(Glu)was measured after 8 weeks of intervention. The biopsy skin tissue was observed under a light microscope. The concentration of AGEs was detected by ELISA. The mRNA expression levels of RAGE,NF-κB p65,IL-6 and TNF-α were detected by RT-qPCR.ResultsUnder the microscope,the skin tissue of the BC group showed structural disorder,thinner thickness,swollen cells,sparse collagen and other obvious pathological changes. The MSC group showed significant improvement compared with the former. Compared with BC group,the levels of Glu,skin AGEs and mRNA expression of the above four factors were decreased in the other three groups(P<0. 05). Compared with MSC group,the above indicators of INS group increased(P<0. 05).ConclusionhUCMSCs can inhibit AGEs/RAGE/NF-κB signaling pathway to a certain ex-tent,which may be one of the mechanisms to improve the"hidden damage"of diabetic skin tissue.
ObjectiveTo observe the effects of human umbilical cord mesenchymal stem cells(h UCMSCs)on AGEs/RAGE/NF-κB signaling pathway in NOD mice skin tissue,and to explore its effect on improving the"hidden damage"of diabetic skin tissue. mechanism.MethodsA total of 43 NOD mice were divided into normal control group(NC,n=12)and diabetic model group(DM,n=31)after 9 weeks of feeding. The model group was further divided into stem cell treatment group(MSC,n=11),insulin treatment group(INS,n=10)and blank control group(BC,n=10). Random blood glucose(Glu)was measured after 8 weeks of intervention. The biopsy skin tissue was observed under a light microscope. The concentration of AGEs was detected by ELISA. The mRNA expression levels of RAGE,NF-κB p65,IL-6 and TNF-α were detected by RT-qPCR.ResultsUnder the microscope,the skin tissue of the BC group showed structural disorder,thinner thickness,swollen cells,sparse collagen and other obvious pathological changes. The MSC group showed significant improvement compared with the former. Compared with BC group,the levels of Glu,skin AGEs and mRNA expression of the above four factors were decreased in the other three groups(P<0. 05). Compared with MSC group,the above indicators of INS group increased(P<0. 05).ConclusionhUCMSCs can inhibit AGEs/RAGE/NF-κB signaling pathway to a certain ex-tent,which may be one of the mechanisms to improve the"hidden damage"of diabetic skin tissue.
引文
[1]林炜栋,钱雄,江万里,等.糖尿病皮肤病理生理改变及其机制的研究.同济大学学报医学版,2012,33:714.
[2]陈向芳,林炜栋,陆树良,等.糖尿病大鼠深二度烫伤创面中成纤维细胞生物学功能的改变.中华医学杂志,2007,87:18121816.
[3]陆树良,青春,谢挺,等.糖尿病皮肤“隐性损害”的机制研究.中华创伤杂志,2004,20:468473.
[4] Brett J,Schmidt AM,Yan S D,et al. Survey of the distribution of a newly characterized receptor for advanced glycation end products in tissues. Am J Pathol,1993,143:16991712.
[5] Clifton DR,Rydkina E,Freeman RS,et al. NFkappa B activation during rickettsia rickettsii infection of endothelial cells involves the activation of catalyticκb kinases ikk alpha and ikk beita and phosphorylationproteolysis of the inhibitor protein i kappa b alpha. Infection&Immunity,2005,73:155165.
[6] Bierhaus A,Humpert PM,Morcos M,et al. Understanding RAGE,the receptor for advanced glycation end products. J Mol Med,2005,83:876886.
[7] Tobonvelasco JC,Cuevas E,Torresramos MA,et al. Receptor for AGEs(RAGE)as mediator of NFkB pathway activation in neuroinflammation and oxidative stress. CNS&Neurological DisordersDrug Targets,2014,13:16151626.
[8] Li H,Fu X. Mechanisms of action of mesenchymal stem cells in cutaneous wound repair and regeneration. Cell Tissue Res,2012,348:371377.
[9] Kwon DS,Gao X,Bo LY,et al. Treatment with bone marrowderived stromal cells accelerates wound healing in diabetic rats.Int Wound J,2008,5:453463.
[10]Khosravi MM,Bagheri M,Solhjou ZH,et al. Stem cell and regenerative medicine:O41:The effect of adipose tissue derived mesenchymal stem cells(admscs)on wound healing of diabetic rat model. 2011.
[11]Tark KC,Hong JW,Kim YS,et al. Effects of human cord blood mesenchymal stem cells on cutaneous wound healing in leprdb mice. Ann Plast Surg,2010,65:565572.
[12]Harrington C,Zagari MJ,Corea J,et al. A cost analysis of diabetic lowerextremity ulcers. Diabetes Care,2000,23:13331338.
[13]Hogan P,Dall T,Nikolov P. Economic costs of diabetes in the US in 2002. Diabetes Care,2008,26:917932.
[14]Gordois A,Scuffham P,Shearer A,et al. The health care costs of diabetic peripheral neuropathy in the US. Diabetes Care,2003,26:17901795.
[15]Shearer A,Scuffham P,Gordois A,et al. Predicted costs and outcomes from reduced vibration detection in people with diabetes in the U. S. Diabetes Care,2003,26:23052310.
[16]陈宾,牛轶雯,谢挺,等.糖尿病大鼠皮肤组织糖代谢紊乱与皮肤神经病变相关性研究.中华烧伤杂志,2011,27:139144.
[17]陈娟,师彦平.糖尿病相关的主要酶靶及其治疗药.中国新药杂志,2003,12:509512.
[18]Goova MT,Li J,Kislinger T,et al. Blockade of receptor for advanced glycation endproducts restores effective wound healing in diabetic mice. Am J Pathol,2001,159:513.
[19]孟庆元,林炜栋,陈向芳. AGERAGE系统与糖尿病足综合征的发病机制及治疗进展.药学服务与研究,2009,9:118121.
[2]陈向芳,林炜栋,陆树良,等.糖尿病大鼠深二度烫伤创面中成纤维细胞生物学功能的改变.中华医学杂志,2007,87:18121816.
[3]陆树良,青春,谢挺,等.糖尿病皮肤“隐性损害”的机制研究.中华创伤杂志,2004,20:468473.
[4] Brett J,Schmidt AM,Yan S D,et al. Survey of the distribution of a newly characterized receptor for advanced glycation end products in tissues. Am J Pathol,1993,143:16991712.
[5] Clifton DR,Rydkina E,Freeman RS,et al. NFkappa B activation during rickettsia rickettsii infection of endothelial cells involves the activation of catalyticκb kinases ikk alpha and ikk beita and phosphorylationproteolysis of the inhibitor protein i kappa b alpha. Infection&Immunity,2005,73:155165.
[6] Bierhaus A,Humpert PM,Morcos M,et al. Understanding RAGE,the receptor for advanced glycation end products. J Mol Med,2005,83:876886.
[7] Tobonvelasco JC,Cuevas E,Torresramos MA,et al. Receptor for AGEs(RAGE)as mediator of NFkB pathway activation in neuroinflammation and oxidative stress. CNS&Neurological DisordersDrug Targets,2014,13:16151626.
[8] Li H,Fu X. Mechanisms of action of mesenchymal stem cells in cutaneous wound repair and regeneration. Cell Tissue Res,2012,348:371377.
[9] Kwon DS,Gao X,Bo LY,et al. Treatment with bone marrowderived stromal cells accelerates wound healing in diabetic rats.Int Wound J,2008,5:453463.
[10]Khosravi MM,Bagheri M,Solhjou ZH,et al. Stem cell and regenerative medicine:O41:The effect of adipose tissue derived mesenchymal stem cells(admscs)on wound healing of diabetic rat model. 2011.
[11]Tark KC,Hong JW,Kim YS,et al. Effects of human cord blood mesenchymal stem cells on cutaneous wound healing in leprdb mice. Ann Plast Surg,2010,65:565572.
[12]Harrington C,Zagari MJ,Corea J,et al. A cost analysis of diabetic lowerextremity ulcers. Diabetes Care,2000,23:13331338.
[13]Hogan P,Dall T,Nikolov P. Economic costs of diabetes in the US in 2002. Diabetes Care,2008,26:917932.
[14]Gordois A,Scuffham P,Shearer A,et al. The health care costs of diabetic peripheral neuropathy in the US. Diabetes Care,2003,26:17901795.
[15]Shearer A,Scuffham P,Gordois A,et al. Predicted costs and outcomes from reduced vibration detection in people with diabetes in the U. S. Diabetes Care,2003,26:23052310.
[16]陈宾,牛轶雯,谢挺,等.糖尿病大鼠皮肤组织糖代谢紊乱与皮肤神经病变相关性研究.中华烧伤杂志,2011,27:139144.
[17]陈娟,师彦平.糖尿病相关的主要酶靶及其治疗药.中国新药杂志,2003,12:509512.
[18]Goova MT,Li J,Kislinger T,et al. Blockade of receptor for advanced glycation endproducts restores effective wound healing in diabetic mice. Am J Pathol,2001,159:513.
[19]孟庆元,林炜栋,陈向芳. AGERAGE系统与糖尿病足综合征的发病机制及治疗进展.药学服务与研究,2009,9:118121.