Objective To investigate the improvement of exosomes(Exo)derived from umbilical cord mesenchymal stem cells(MSCs)carrying miR-296-3p on blood lipid metabolism and immune function in gestational diabetes melli-tus rats.Methods After transfection of MSCs with miR-296-3p-NC and miR-296-3p mimics,Exo was extracted to obtain MSCs-Exo-miR-NC and MSCs-Exo-miR-296-3p,respectively.RT-qPCR was used to detect the expression of miR-296-3p;Pregnant mice were prepared and induced to establish a GDM model by tail vein injection of strep-tozotocin.Then they were randomly grouped into a model group,MSCs-Exo-miR-NC group,and MSCs-Exo-miR-296-3p group with 10 each.In addition,10 pregnant mice were prepared as the control group.The weight of mother and fetal mice,four items of blood lipids and immune organ index in each group were measured after intervention with MSCs-Exo-miR-NC and MSCs-Exo-miR-296-3p.ELISA was applied to measure the level of serum immuno-globulin and immune inflammatory factors.Flow cytometry technology was applied to detect the immune function of peripheral blood T lymphocytes of rats in various groups,and their CD4+and CD8+values were compared.Results Compared with the control group,the maternal rat weight and fetal rat weight,serum triglycerides(TG),total cho-lesterol(TC),low-density lipoprotein(LDL-C),IL-6 and IL-8 levels and CD8+ratio of rats in model group in-creased(P＜0.05),the spleen and thymus index,serum high-density lipoprotein(HDL-C),IgA,IgG,IgM,IL-2 and IL-10 levels and CD4+ratio decreased(P＜0.05).Compared with the model group,the maternal rat weight and fetal rat weight,serum TG,TC,LDL-C,IL-6 and IL-8 level,and CD8+ratio of rats from MSCs-Exo-miR-NC group and MSCs-Exo-miR-296-3p group decreased(P＜0.05),the spleen and thymus index,serum HDL-C,IgA,IgG,IgM,IL-2 and IL-10 levels and CD4+ratio all increased(P＜0.05);MSCs-Exo-miR-296-3p had a stronger effect on various indicators of GDM rats(P＜0.05).Conclusions Exo derived from MSCs carrying miR-296-3p improves blood lipid metabolism and immune function of GDM rats.

Once pulp necrosis or apical periodontitis occurs on immature teeth, the weak root and open root apex are challenging to clinicians. Berberine (BBR) is a potential medicine for bone disorders, therefore, we proposed to apply BBR in root canals to enhance root repair in immature teeth. An in vivo model of immature teeth with apical periodontitis was established in rats, and root canals were filled with BBR, calcium hydroxide or sterilized saline for 3 weeks. The shape of the roots was analyzed by micro-computed tomography and histological staining. In vitro, BBR was introduced into stem cells from apical papilla (SCAPs). Osteogenic differentiation of stem cells from apical papilla was investigated by alkaline phosphatase activity, mineralization ability, and gene expression of osteogenic makers. The signaling pathway, which regulated the osteogenesis of SCAPs was evaluated by quantitative real time PCR, Western blot analysis, and immunofluorescence. In rats treated with BBR, more tissue was formed, with longer roots, thicker root walls, and smaller apex diameters. In addition, we found that BBR promoted SCAPs osteogenesis in a time-dependent and concentration-dependent manner. BBR induced the expression of β-catenin and enhanced β-catenin entering into the nucleus, to up-regulate more runt-related nuclear factor 2 downstream. BBR enhanced root repair in immature teeth with apical periodontitis by activating the canonical Wnt/β-catenin pathway in SCAPs.
Animals ; Berberine ; pharmacology ; Cell Differentiation ; drug effects ; Dental Papilla ; Male ; Osteogenesis ; drug effects ; Periapical Periodontitis ; therapy ; Rats ; Stem Cells ; cytology ; drug effects ; metabolism ; Wnt Signaling Pathway ; drug effects ; Wnt3A Protein ; genetics ; metabolism ; X-Ray Microtomography