Therapeutic Effects Of RhMG53 Combination With HUC-MSCs In Alzheimer’s Mice

Alzheimer’s disease is a neurodegenerative disease closely related to age. AD patients showed an increasing trend with the aging of our society in recent years. At present, the elucidation of the pathogenesis and the improvement of the treatment methods are still difficult problems in medical field. Stem cell transplantation is a new therapy for disease, which brings hope for the treatment of AD. Improving the survival ability of transplanted stem cells and promoting more stem cell migration into the brain tissue play an important role in improving the effect of stem cell transplantation. Protein Mitsugumin53 is a newly discovered TRIM family muscle specific protein, which plays a key role in cell membrane damage repair and multiple acute organ injury. But the protective effect of human Mitsugumin53 protein recombinant was not reported on umbilical cord mesenchymal stem human cells oxidative damage and AD chronic pathological state. Aim:In vitro,the protective effect of rh MG53 on oxidative damage of h UC-MSCs were studied by H2O2 oxidative damage h UC-MSCs model.In vivo, the effects of rh MG53 combined with h UC-MSCs transplantation on the treatment of APP+ transgenic model mice were studied in order to understanding the function of MG53 protein and provide a scientific basis for the therapeutic effect of AD. Methods:1. Protective effects of rh MG53 on oxidative damage in h UC-MSCs in vitroh UC-MSCs were isolated from healthy human umbilical cord by tissue mass cultural method and H2O2 damage model were established using P3 h UC-MSCs. Effect of rh MG53 protein on cell proliferation were detected by CCK-8 method, and the experiment were divided into NC group, MG53 group, H2O2 group, pretreatment MG53 group(Pre group).Cell migration were observed by Transwell, cell cycle and apoptosis were analyzed by flow cytometry, cell senescence were detected by β-galactosidase staining, cell SOD activity and MDA content were detected by spectrophotometric.2. Therapeutic effects of rh MG53 combination with h UC-MSCs in Alzheimer’s mice4 month old APP+ transgenic Alzheimer’s disease model mouse were divided into APP+ group, h UC-MSCs group, rh MG53 group, rh MG53 combined with h UC-MSCs group(combined group). After a week, the migration of h UC-MSCs in the mouse brain were detected by immunohistochemistry. After two weeks, mouse exercise capacity were detected by treadmill and after four weeks, the learning and memory ability were measured by morris water maze test. Activity of superoxide dismutase(SOD) and concentration of Malonaldehyde(MDA) in the serum were quantified by microplate spectrophotometer and expression of senescence related factors(sirt2、PCNA、P16、P53) were quantified by q RT-PCR and Western blot, neural markers(DCX, Nestin, NSE) were detected by immunohistochemistry.Results:1. P3 h UC-MSCs adherent growth, the cells morphology were spindle shaped spiral, smooth and clear, but cell adhesion were decreased significantly and the cells surface were not smooth after H2O2 treatment. it has concentration and time dependent damage effect on h UC-MSCs, so we use these conditions that h UC-MSCs were cultured 16 h with 200μM H2O2 as the optimum condition for the oxidative damage model. rh MG53 has a concentration dependent enhancement effect on cell proliferation and 30μg/m L were used as experimental concentration. Compared with NC group, MG53 group cell proliferation and migration ability were increased significantly, cells proportion in S phase had not significant difference, cell senescence and apoptosis rate were reduced significantly, cell SOD activity were increased significantly, MDA content decreased significantly; H2O2 group cell proliferation and migration ability were reduced significantly, cell proportion in S phase were reduced significantly, cell senescence and apoptosis rate were increased significantly, cell SOD activity were increased significantly, MDA content decreased significantly.Compared with H2O2 group, Pre group cell proliferation and migration ability were increased significantly,cells proportion in S phase had not significant difference,cell senescence and apoptosis rate were reduced significantly, cell SOD activity were increased significantly, MDA content decreased significantly.2. There were 40 APP+ mice identified by PCR. Compared with h UC-MSCs group, the number of h UC-MSCs in combined group mouse brain were increased significantly after a week. After two weeks, movement ability of MG53 group and combined group were increased significantly. Four weeks after transplantation, compared with the APP+ group, in other groups the escaping latency were shorter significantly and the number of crossing platform were increased significantly and the time spent in the platform quadrant were longer significantly; SOD activity were increased and MDA content were decreased significantly in the serum; Sirt1, Sirt2, PCNA gene expression increased and P16, P21, P53 gene expression decreased significantly in the m RNA and protein level, h UC-MSCs group and combined group neural marker DCX and Nestin increased in mouse brain, NSE expression changes were not significant. Compared with h UC-MSCs group, combined group escaping latency were shorter significantly and SOD activity increased and MDA content decreased significantly in the serum, Sirt1, Sirt2, PCNA gene expression increased and P16, P21, P53 gene expression decreased significantly in the m RNA and protein level.Conclusion:1. rh MG53 increased the survival and migration and enhanced the antioxidant capacity of h UC-MSCs, protects cells against H2O2 oxidative damage.2. rh MG53 improve the survival and migration of h UC-MSCs in APP+ mice, improve the learning and cognitive ability, and promote the therapeutic effect of h UC-MSCs on APP+ mice.