Evaluation Of Therapeutic Effect Of MitoQ Pretreated Adipose Derived Mesenchymal Stem Cells On Canine Diabetes Mellitus

Currently,the diagnosis of Diabetes mellitus（DM）is no longer limited to humans,but has also become a more common chronic metabolic disease in companion animals such as canines,characterized by persistent hyperglycemia and insulin deficiency.As the disease progresses,it mostly leads to complications,which will greatly increase the disability and mortality rates.However,the current treatment for canine diabetes is relatively single,mostly insulin injections with oral hypoglycemic drugs,as well as attention to diet and exercise,which has certain drawbacks,such as inability to accurately grasp the amount of insulin injections,the difficulty of oral administration of drugs in canines with some hepatotoxicity and nephrotoxicity,the high cost of treatment and the inability of canines to give effective feedback,etc.Therefore,in clinical treatment,there is an urgent need for an effective,inexpensive,and safe method to provide a new direction for the treatment of canine diabetes.Mesenchymal stem cells（MSCs）are the most widely used adult stem cells in chinical applications in recent years.In addition to the mutal feature of stem cells such as multi-differentiation into bone,cartilage,adipose,nerve tissue cells and self-renewal,they also have characteristics such as hematopoietic support and immune regulation,which tremendously accelerate the advancement of MSCs in clinical applications.MSCs have also shown good therapeutic potential in diabetes and related diseases.Compared with other types of MSCs,Adipose-derived mesenchymal stem cells（ADMSCs）have become the seed cells with enormous capacity for treatments because of their convenient gain to materials and absence of moral and ethical controversies,however,ADMSCs still have some shortcomings,such as easy senescence during in vitro culture and short duration of action after cell transplantation into the body.However,ADMSCs have some shortcomings,such as easy aging during in vitro culture and short duration of action after cell transplantation into vivo.Accordingly,it is crucial to discover a methodology to scale up the curing quality of ADMSCs on diabetes.Mitoquionl Mesylate（Mito Q）is a powerful antioxidant that has been proven in numerous clinical trials to delay cellular senescence and reduce cellular damage.Formerly studies have also make clear that Mito Q can play a good role in the treatment of neurodegenerative and metabolic diseases by reducing oxidative stress and mitochondrial dysfunction.Consequently,in this research,we performed pretreatment of ADMSCs with Mito Q,and conducted experiments on the effects of Mito Q on the growth,proliferation,migration,antioxidant and anti-aging abilities of ADMSCs,as well as the enhancement of the therapeutic effects of Mito Q pretreatment of ADMSCs on diabetes in mice and canines,obtained the following results:（1）Pretreatment of ADMSCs with Mito Q did not change the cell morphology,but remarkable facilitated the growth acceleration,proliferation ability and migration capability of ADMSCs.In addition,Mito Q also promoted the enhancement of antioxidant capacity of ADMSCs,as well as delayed cell senescence and apoptosis.（2）Under the combined effect of high-fat diet（HFD）and streptozotocin（STZ）injection,the clinical indexes of the animal models were changed,both mice and canines showed symptoms such as lighter body weight,elevated blood glucose level,depression,coarse coat,and extremely heavy urine taste,dilute stool and blood stool.The fasting blood glucose of mice and random blood glucose of canines were higher than 11.1 mmol/L.The diabetes mellitus models of mice and canines were successfully established.（3）Cell transplantation was performed in mice and canines,and the number of cells transplanted in mice was 2×10⁶ cells and in dogs was 1×10⁷cells,and the results showed that ADMSCs effectively reduced diabetes induced hyperglycemia,and Mito Q could further promote the recovery of blood glucose level in the body.In addition,Mito Q was able to promote more significant recovery of ADMSCs from other clinical symptoms induced by diabetes,such as reduced body weight,increased feeding and water intake,and depression.（4）At the serological level,ADMSCs could improve liver and lipid metabolism indexes,and promote the reduction of liver and lipid metabolism disorders in diabetic animal models,and Mito Q could further promote the therapeutic effects of ADMSCs;Metabolomic analysis of canine serum showed that Mito Q was able to greatly promote the therapeutic effect of ADMSCs on canine diabetes by upregulating the expression of antioxidant-related metabolites and restoring them to normal levels.（5）From the histological level,the levels of SOD,H₂O₂,MDA and CAT in pancreatic tissues were examined for the related indexes of oxidative stress,and it was seen that Mito Q-ADMSCs could effectively promote the improvement of antioxidant-related functions in diabetic model animals and promote the repair of oxidative damage in tissues;after the related staining of pancreas and liver,it was seen that ADMSCs had a beneficial effect on pancreatic and Mito Q pretreatment of ADMSCs can further promote this therapeutic effect and restore them to the level of the initial stage of diabetes or even to the normal state,indicating that Mito Q pretreatment of ADMSCs can further improve the therapeutic effect of diabetes.Thus,the use of MSCs in clinical treatment is greatly facilitated.In the present study,a high-fat diet（HFD）combined with STZ was used to induce diabetes in mice and canines,and the therapeutic effects of Mito Q pretreated ADMSCs on diabetes were evaluated by monitoring clinical parameters,serological levels and histological levels.The results showed that when diabetes occurs,the animal organism will experience disruptions in the endocrine system due to elevated blood glucose,which in turn leads to the occurrence of tissue damage,disorders of liver and lipid metabolism,and deepening of oxidative stress,etc.By pretreating ADMSCs with Mito Q can well improve clinical indices,reduce the occurrence of oxidative stress and metabolic disorders,and repair tissue damage,thus providing a better opportunity for MSCs to be applied to diabetes and its related diseases.