Please wait a minute...
浙江大学学报（医学版）
浙江大学学报（医学版）  2011, Vol. 40 Issue (6): 630-640    DOI: 10.3785/j.issn.1008-9292.2011.06.010
论著     
Wnt/β-catenin信号通路对间充质干细胞衰老的影响及其作用机制
项晓霞，陈律，王骏浩，张余斌，张大勇
浙江大学城市学院医学与生命科学学院，浙江 杭州 310015
Role of Wnt/β-catenin signaling in aging of mesenchymal stem cells of rats
XIANG Xiao-xia,CHEN Lü,WANG Jun-hao,ZHANG Yu-bin,ZHANG Da-yong
School of Medicine and Life Science,Zhejiang University City College,Hangzhou 310015,China
 全文: PDF(4665 KB)   HTML ( ) 
摘要： 目的：研究不同年龄血清对Wnt/β-catenin信号通路激活水平的影响，探讨Wnt/β-catenin信号通路对间充质干细胞（MSCs）衰老、增殖、存活能力的作用及其机制。
方法：提取年轻（8~12 周）及老年（64～72 周）SD大鼠血清，实验分为4组，分别为年轻血清（Young rat serum,YRS）组，YRS+Wnt 3a组、老年血清（Old rat serum,ORS）组和ORS+DKK1组。免疫细胞化学染色和Western blotting检测胞浆、胞核β-catenin的表达。SA-β-半乳糖苷酶染色检测各组细胞衰老，MTT法检测细胞增殖，AO/EB染色法检测MSCs存活和凋亡。为研究Wnt/β-catenin信号通路导致MSCs衰老的作用机制，免疫细胞化学染色和Western blotting检测γ-H2A.X和p53蛋白表达，RT-PCR法检测p53、p21 mRNA表达。
结果：ORS组β-catenin表达较YRS组明显增强，尤其是胞核，出现β-catenin聚集现象，而ORS+DKK1组β-catenin表达减弱。与YRS组相比，YRS+Wnt 3a组细胞SA-β-半乳糖苷酶染色阳性细胞数显著增多（P<0.01），细胞增殖和存活能力减弱。而ORS+DKK1组与ORS组相比，SA-β-半乳糖苷酶染色阳性细胞数显著减少（P<0.01），细胞增殖和存活能力增强。免疫细胞化学染色、Western blotting和RT-PCR检测结果显示，ORS组内γ-H2A.X、p53、p21表达较YRS组明显增强，而在ORS内加入DKK1后，γ-H2A.X、p53、p21表达较ORS组明显减弱。
结论：ORS培养可以激活MSCs内Wnt/β-catenin信号通路。Wnt/β-catenin信号通路对MSCs衰老发挥重要调控作用。DNA损伤反应和p53/p21途径是Wnt/β-catenin信号通路导致MSCs衰老的重要介导因素。
关键词： 间质干细胞/病理学;  细胞增殖;  信号传导;  Wnt/&beta;  -catenin信号通路;  骨髓间充质干细胞;  血清;  衰老;  DNA损伤反应;  p53/p21途径    
Abstract: Objective： To investigate the role of Wnt/β-catenin signaling in aging of mesenchymal stem cells (MSCs) of rats.
Methods： Serum samples were collected from young (8~12 w) and aged (64~72 w) SD rat.Four experiment groups were assigned:young rat serum (YRS),YRS+Wnt 3a,old rat serum (ORS) and ORS + DKK1 groups.Immunofluorescence and Western blotting were used to detect the expression of intracellular β-catenin.The senescence-associated changes were examined with SA-β-galactosidase staining.The proliferation ability was tested by MTT assays.The survived and apoptotic cells were determined by AO/EB staining.The expressions of γ-H2A.X and p53 protein were detected by immunofluorescence and Western blotting.RT-PCR was used to detect the expression of p53 and p21 mRNA.
Results： Compared with the YRS group,the intracellular expression of β-catenin in the ORS group was significantly increased,especially in the nuclei of MSCs.After treatment of DKK1 in ORS,the β-catenin expression was reduced.The number of SA-β-galactosidase positive MSCs was significantly higher in the YRS+Wnt 3a group than that in the YRS group (P<0.01),and the proliferative and survival ability of MSCs was significantly decreased in the YRS+Wnt 3a group.The number of SA-β-galactosidase positive MSCs in the ORS+DKK1 group was significantly decreased compared with that in ORS group (P<0.01),and the proliferative and survival ability of MSCs was significantly increased in the ORS+DKK1 group.The expression of γ-H2A.X,p53 and p21 was markedly increased in the ORS group than that in YRS group,however,after treatment with Wnt/β-catenin signaling inhibitor DKK1,the expression of γ-H2A.X,p53 and p21 was significantly decreased compared with that in the ORS group.
Conclusions： Results suggest that the Wnt/β-catenin signaling is activated in the MSCs cultured with ORS and excessive activation of Wnt/β-catenin signaling can promote MSCs aging.The DNA damage response and p53/p21 pathway may be main mediators of MSC aging induced by excessive activation of Wnt/β-catenin signaling.
Key words: Mesenchymal stem cells/pathol    Cell proliferation    Signal transduction    Wnt/β-catenin signaling    Mesenchymal stem cells (MSCs)    Aging    serum    DNA damage response    p53/p21 pathway
出版日期: 2011-11-25
服务　　
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章　　

引用本文:

项晓霞;陈律;王骏浩;张余斌;张大勇. Wnt/β-catenin信号通路对间充质干细胞衰老的影响及其作用机制[J]. 浙江大学学报（医学版）, 2011, 40(6): 630-640.

链接本文:

https://www.zjujournals.com/med/CN/Y2011/V40/I6/630

[1] 严敏,汪旭,郭锡汉. 早老蛋白1功能缺失性突变导致家族性阿尔茨海默病的研究进展[J]. 浙江大学学报（医学版）, 2020, 49(4): 487-499.
[2] 黄耀凭,杨凤,周天华,谢珊珊. Hippo信号通路及其在消化系统肿瘤中的作用研究进展[J]. 浙江大学学报（医学版）, 2020, 49(1): 35-43.
[3] 徐亦鸣,应可净. 中性粒细胞胞外诱捕网与肿瘤相关研究进展[J]. 浙江大学学报（医学版）, 2020, 49(1): 107-112.
[4] 方娟,潘志成,郭晓纲. INK4基因座中反义非编码RNA调控细胞增殖与凋亡影响动脉粥样硬化的研究进展[J]. 浙江大学学报（医学版）, 2020, 49(1): 113-117.
[5] 王燚锋, 王志萍. 内源性信号通路在神经元轴突再生中的功能和机制研究[J]. 浙江大学学报（医学版）, 2020, 49(1): 82-89.
[6] 张大勇,林九洲,王雅艳,许珊,罗成专,蔡佳烨,蒋学范,潘建平. 白藜芦醇通过保护线粒体功能延缓骨髓间充质干细胞衰老[J]. 浙江大学学报（医学版）, 2019, 48(6): 617-624.
[7] 徐美娇,王一枫. 二苯乙烯苷可抑制中波紫外线诱导的人皮肤成纤维细胞光老化[J]. 浙江大学学报（医学版）, 2019, 48(6): 625-630.
[8] 刘婧雯,杨兴莲,沈凯莉,曾玲晖,孙燕. 氯氧喹通过下调Rho/Rho激酶信号通路抑制乳腺癌细胞转移[J]. 浙江大学学报（医学版）, 2019, 48(6): 631-637.
[9] 王青梅, 徐千姿, 魏安怡, 陈世硕, 张翀, 曾玲晖. 大剂量维生素C通过减少糖酵解和蛋白质合成抑制乳腺癌细胞增殖[J]. 浙江大学学报（医学版）, 2019, 48(3): 296-302.
[10] 朱紫菱, 谈静, 邓红. 肿瘤细胞膜/质蛋白转位入核研究进展[J]. 浙江大学学报（医学版）, 2019, 48(3): 318-325.
[11] 邵佳乐,李志忠,周建,李凯,秦荣,陈克明. 低频脉冲电磁场通过IGF-1R/NO信号通路促进大鼠颅骨成骨细胞成熟及矿化[J]. 浙江大学学报（医学版）, 2019, 48(2): 158-164.
[12] 赵世浩,张雪,柯越海. 细胞衰老与特发性肺纤维化的相关性研究进展[J]. 浙江大学学报（医学版）, 2019, 48(1): 111-115.
[13] 梁刚, 牛育苗, 李一涵, 魏安怡, 董静尹, 曾玲晖. 雷帕霉素在大鼠局灶性脑缺血再灌注后24 h给药对脑损伤的保护作用[J]. 浙江大学学报（医学版）, 2018, 47(5): 443-449.
[14] 朱锋,范苗,徐孜惟,蔡依廷,陈益臻,余双,曾玲晖. 雷帕霉素对帕金森病小鼠的保护作用[J]. 浙江大学学报（医学版）, 2018, 47(5): 465-472.
[15] 钱波,张彦玲,莫绪明. 先天性食管闭锁相关转录因子及信号通路研究进展[J]. 浙江大学学报（医学版）, 2018, 47(3): 239-243.