rhMG53蛋白联合 hUC－MSCs 移植对阿尔茨海默鼠的治疗作用

摘要

目的：研究 rhMG53蛋白联合人脐带间充质干细胞（hUC－MSCs）移植对阿尔茨海默模型鼠（APP 转基因小鼠）的治疗作用。方法：40只 APP＋小鼠随机分为 APP＋组、hUC－MSCs 组、rhMG53组和 rhMG53联合 hUC－MSCs组（联合组），分组处理1周后采用免疫组化法检测小鼠脑内 hUC－MSCs 的迁移率，3周后采用 Morris 水迷宫测试各组小鼠的学习和记忆能力，同时检测小鼠血清中 SOD 的活性和 MDA 的含量，并采用 qRT－PCR 和 Western blot 检测小鼠脑组织中衰老相关基因（sirt2、pCNA、p16、p53）的表达。结果：APP＋组和 rhMG53组未见 hUC－MSCs 迁移，hUC－MSCs 组和联合组存在 hUC－MSCs 迁移，且联合组迁移细胞更多（P ＜0．05）。 hUC－MSCs 移植可以使 APP＋小鼠的逃避潜伏期缩短，穿越平台次数增加以及平台所在象限停留时间增加（P ＜0．05）；hUC－MSCs 移植和 rhMG53均可使APP＋小鼠血清 SOD 活性增加，MDA 含量下降，且二者联用对 MDA 的影响有协同作用（P ＜0．05）；hUC－MSCs 移植和 rhMG53均可使 APP＋小鼠脑组织 sirt2和 pCNA mRNA 和蛋白的相对表达量升高，且二者联用对多数衰老相关基因表达的影响有交互作用（P ＜0．05）。结论：外源 rhMG53蛋白可以提高小鼠脑内 hUC－MSCs 的迁移，促进 hUC－MSCs 对阿尔茨海默鼠的治疗作用。%Aim: To research the effects of rhMG53 combination with hUC-MSCs transplantation on Alzheimer′s dis-ease mice.Methods: Forty APP+ mice were randomly allocated into four groups : APP+ group, hUC-MSCs group, rh-MG53 group and rhMG53 combined with hUC-MSCs group.The migration of hUC-MSCs was quantified by immunohisto-chemistry at one week after transplantation .And the learning and memory ability was measured by Morris water maze test at three weeks after transplantation .The activity of superoxide dismutase (SOD) and the concentration of malonaldehyde (MDA) in the serum were also quantified.qRT-PCR and Western blot were used to detect the expressions of senescence related factors(sirt2,pCNA,p16,p53).Results: No migration of hUC-MSCs was found in APP + group or rhMG53 group, while it was observed in the hUC-MSCs group and the combined group , and the amount of migration cells was higher in the combined group(P <0.05).hUC-MSCs transplantation decreased the escaping latency , increased the times of crossing platform and the time spent in the platform quadrant (P <0.05).hUC-MSCs transplantation and rhMG53 could both in-crease the activity of SOD and decrease the content of MDA in the serum of APP+ mice, and rhMG53 combined with hUC-MSCs transplantation had synergistic effect on MDA (P <0.05).hUC-MSCs transplantation and rhMG53 could both in-crease the expressions of sirt2 and pCNA mRNA and protein in the brain tissue of APP+ mice, and rhMG53 combined with hUC-MSCs transplantation had synergistic effect on the expression of most aging genes (P <0.05).Conclusion: rhMG53 can improve the migration of hUC-MSCs in brain tissue of APP+ mice, and promote the therapeutic effect of hUC -MSCs on APP+ mice.