郑州大学学报（医学版）
鄭州大學學報（醫學版）
정주대학학보（의학판）
JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES)
2015年
1期
25-29
,共5页
崔渊博%马珊珊%姚宁%渠瑞娜%王欣欣%邢衢%孟楠%杨波%关方霞
崔淵博%馬珊珊%姚寧%渠瑞娜%王訢訢%邢衢%孟楠%楊波%關方霞
최연박%마산산%요저%거서나%왕흔흔%형구%맹남%양파%관방하
干细胞移植%衰老%阿尔茨海默病%衰老相关基因
榦細胞移植%衰老%阿爾茨海默病%衰老相關基因
간세포이식%쇠로%아이자해묵병%쇠로상관기인
stem cell transplantation%aging%Alzheimer’s disease%aging related gene
目的：研究人脐带间充质干细胞（hUC－MSCs）静脉移植对APP＋转基因鼠衰老的调控作用，并从衰老相关基因角度探讨MSCs延缓衰老的机制。方法：采用PCR技术对老年痴呆APP＋转基因鼠进行分子鉴定；将传至第3代的hUC－MSCs细胞密度调整为1×106 mL －1后进行尾静脉移植。移植hUC－MSCs 3周后，通过Morris水迷宫实验从行为学水平检测干细胞移植对APP ＋转基因鼠的延缓衰老作用（检测小鼠的学习记忆能力），然后采用荧光定量PCR和Western blot法检测hUC－MSCs移植后小鼠脑及肝脏组织中与衰老相关的基因及蛋白[沉默信息调节因子2（Sirt2）、增殖细胞核抗原（PCNA）、 p16、p21、p53]表达的变化。结果：该实验共鉴定出12只APP＋转基因鼠。与对照组相比，移植组小鼠逃避潜伏期时间明显缩短、穿越平台次数增加、在平台所在象限停留的时间延长，差异具有统计学意义（P均＜0．05）。移植组APP＋转基因鼠的脑及肝脏组织中PCNA和Sirt2基因的mRNA和蛋白相对表达量升高， p21及p53基因相对表达量降低（P均＜0．05），而p16基因表达变化不明显（P＞0．05）。结论：APP＋转基因鼠经hUC－MSCs静脉移植治疗，学习记忆能力显著提高，延缓衰老作用明显，涉及衰老标志基因p21、p53、PCNA、Sirt2等表达调控改变，这些衰老标志基因之间相互联系，可能构成细胞衰老的基因调控链。
目的：研究人臍帶間充質榦細胞（hUC－MSCs）靜脈移植對APP＋轉基因鼠衰老的調控作用，併從衰老相關基因角度探討MSCs延緩衰老的機製。方法：採用PCR技術對老年癡呆APP＋轉基因鼠進行分子鑒定；將傳至第3代的hUC－MSCs細胞密度調整為1×106 mL －1後進行尾靜脈移植。移植hUC－MSCs 3週後，通過Morris水迷宮實驗從行為學水平檢測榦細胞移植對APP ＋轉基因鼠的延緩衰老作用（檢測小鼠的學習記憶能力），然後採用熒光定量PCR和Western blot法檢測hUC－MSCs移植後小鼠腦及肝髒組織中與衰老相關的基因及蛋白[沉默信息調節因子2（Sirt2）、增殖細胞覈抗原（PCNA）、 p16、p21、p53]錶達的變化。結果：該實驗共鑒定齣12隻APP＋轉基因鼠。與對照組相比，移植組小鼠逃避潛伏期時間明顯縮短、穿越平檯次數增加、在平檯所在象限停留的時間延長，差異具有統計學意義（P均＜0．05）。移植組APP＋轉基因鼠的腦及肝髒組織中PCNA和Sirt2基因的mRNA和蛋白相對錶達量升高， p21及p53基因相對錶達量降低（P均＜0．05），而p16基因錶達變化不明顯（P＞0．05）。結論：APP＋轉基因鼠經hUC－MSCs靜脈移植治療，學習記憶能力顯著提高，延緩衰老作用明顯，涉及衰老標誌基因p21、p53、PCNA、Sirt2等錶達調控改變，這些衰老標誌基因之間相互聯繫，可能構成細胞衰老的基因調控鏈。
목적：연구인제대간충질간세포（hUC－MSCs）정맥이식대APP＋전기인서쇠로적조공작용，병종쇠로상관기인각도탐토MSCs연완쇠로적궤제。방법：채용PCR기술대노년치태APP＋전기인서진행분자감정；장전지제3대적hUC－MSCs세포밀도조정위1×106 mL －1후진행미정맥이식。이식hUC－MSCs 3주후，통과Morris수미궁실험종행위학수평검측간세포이식대APP ＋전기인서적연완쇠로작용（검측소서적학습기억능력），연후채용형광정량PCR화Western blot법검측hUC－MSCs이식후소서뇌급간장조직중여쇠로상관적기인급단백[침묵신식조절인자2（Sirt2）、증식세포핵항원（PCNA）、 p16、p21、p53]표체적변화。결과：해실험공감정출12지APP＋전기인서。여대조조상비，이식조소서도피잠복기시간명현축단、천월평태차수증가、재평태소재상한정류적시간연장，차이구유통계학의의（P균＜0．05）。이식조APP＋전기인서적뇌급간장조직중PCNA화Sirt2기인적mRNA화단백상대표체량승고， p21급p53기인상대표체량강저（P균＜0．05），이p16기인표체변화불명현（P＞0．05）。결론：APP＋전기인서경hUC－MSCs정맥이식치료，학습기억능력현저제고，연완쇠로작용명현，섭급쇠로표지기인p21、p53、PCNA、Sirt2등표체조공개변，저사쇠로표지기인지간상호련계，가능구성세포쇠로적기인조공련。
Aim: To research the anti-aging effects of human umbilical cord mesenchymal stem cells ( hUC-MSCs ) transplantation on transgenic APP +mice and to investigate the anti-aging mechanisms in terms of age-related genes. Meth-ods:APP+transgenic mice were identified by PCR. hUC-MSCs were isolated in vitro sterilely. At the third passage , sin-gle cell suspension at 1 ×106 mL-1 was obtained and transplanted by tail venous pathway in the transplantation group. The anti-aging effects on transgenic APP +mice were detected by Morris water maze test at three weeks after transplantation to study the learning and memory ability of the mice. qRT-PCR and Western blot were performed to test the change of mRNA and protein expression of age-related gene in the brain and liver of the APP +mouse, including p16, p21, p53, silence in-formation regulator 2(Sirt2), and proliferating cell nuclear antigen (PCNA).Results:12 APP+mice were identified total-ly.Compared with control group, the escaping latency was significantly shorter (P<0.05), the number of crossing plat-form quadrant increased and the time of crossing platform quadrant was significantly longer in the transplantation group (P<0.05).What’s more, the expressions of Sirt2 and PCNA were increased, and p21, p53 but not p16 decreased in the brain and liver tissue of transplanted APP +mice(P<0.05).Conclusion:The learning and memory ability and anti-aging effect are increased significantly in APP +mice after hUC-MSCs tansplantation. The underlying mechanism is partly due to the change of aging related gene such as p 21, p53, PCNA and Sirt2, which may interact with each other and constitute the gene regulation chain of cell aging.
