| PurposeIn this study,a pregnant mouse model of GDM was established to investigate the induction of human placenta-derived mesenchymal stem cells(hPDMSCs)into islet-like cells(ILCs)under in vitro conditions and feasibilite and explore the relationship between adiponectin(APN),tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6).It is confirmed that human placenta-derived mesenchymal stem cells and their induced islet-like cells in vitro can improve insulin resistance(IR),lower blood sugar,and have a clear therapeutic effect on gestational diabetes.Treatment lays the theoretical foundation.MethodAfter explaining the placenta use to the mother and her family members,and seeking their consent,the healthy placental tissue of the full-term cesarean section was collected.Mesenchymal stem cells were extracted from placental tissues and an appropriate amount of hPDMSCs were induced into pancreatic islet cells in vitro.Diphenylthiocarbazone staining(DTZ)was used to specifically identify the induced islet-like cells.Ten female rats were fed normally,without any treatment,and were set as group A;40 GDM pregnant rat models established by the combination of high-fat and high-sugar feeding + streptozotocin(STZ)injection were set as B and C,D,E 4 experimental groups.After successful modeling,the settings were as follows:Group B:GDM mice without intervention group(10 mice),Group C:GDM mice saline intervention group(10 mice),Group D:GDM mice placental mesenchymal stem cell intervention group(10 mice)),E group:GDM rat islet-like cell intervention group(10 mice),and then treated accordingly.On the 6th,12 th and 18 th days of pregnancy,the fasting blood glucose of each group of pregnant rats was measured,and the pregnant rats were gestated to 18 days(D18),and 5 groups of pregnant rats were given cesarean section to obtain the placenta of each group of pregnant rats The relative expressions of adiponectin,tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)mRNA in the placenta were detected by RT-time PCR technology,and their statistical significance was analyzed using statistical software.Result1.The hPDMSCs were cultured and passaged until they were passed to the third generation hPDMSCs.It was observed under the microscope that the cells grew well and the cells were pure.The surface antigen expression of CD44(+)and CD105(+)was identified by flow cytometry and confirmed as hPDMSCs.2.It was observed under a microscope that the cells after orientation induction changed from circular to spindle-shaped,moved closer to the center and clustered together.After diphenylhydantoin staining,it was reddish brown under the microscope and confirmed to be islet-like cells.3.The criteria for successful GDM rat modeling were:blood glucose> 16.7 mmol / L.Fifty pregnant rats were divided into A,B,C,Dand E5 groups of 10 rats each,which were the normal pregnant rats group,the GDM rats without intervention group,the GDM rats saline intervention group,and the GDM rats human placenta mesenchymal cell intervention group and GDM mouse islet-like cell intervention group.On the 6th,12 th,and 18 th days of pregnancy,the fasting blood glucose of pregnant rats in each group was measured.At the same time point,the blood glucose of GDM pregnant rats in groups B,C,Dand E was significantly higher than that of normal pregnant rats in group A.The differences were:Statistical significance(P <0.05);there was no significant difference in blood glucose between group B(non-intervention group with GDM mice)and group C(normal saline intervention group with GDM mice)(P> 0.05);Group D(GDM rat placental mesenchymal stem cell intervention group),group E(GDM rat islet-like cell intervention group)compared with groups B and C,their blood glucose decreased significantly,the difference was statistically significant(P <0.05);Compared with group D,the blood glucose drop was more significant on the 12 th and 18 th day of pregnancy in group E,and the difference was statistically significant(P <0.05).Intra-group comparison of group D and E,pregnancy Blood glucose decreased on the 12 th day compared with the 6th day of pregnancy,and the difference was statistically significant(P <0.05).There was no significant difference in blood glucose on the 18 th and 12 th days of pregnancy(P> 0.05).Compared with group A,the relative expression of adiponectin mRNA in the placenta tissue of groups B and C was significantly decreased,and the relative expression of TNF-α and IL-6 mRNA was significantly increased,and the difference was statistically significant(P <0.05);There was no significant difference in the relative expression of adiponectin,TNF-α,and IL-6 mRNA in the placental tissues of group B and group C(P> 0.05);compared with group D,group E,group B and group C,The relative expression of adiponectin mRNA in the placenta increased,and the relative expression of TNF-α and IL-6 mRNA decreased.The difference was statistically significant(P <0.05).Compared with group D,group E had lipids in group E.The relative expression level of adiponectin mRNA was higher than that in group D,while the relative expression levels of TNF-α and IL-6 mRNA were lower than those in group D,and the difference was statistically significant(P <0.05).Conclusion1.Both human placenta-derived(hPDMSCs)mesenchymal stem cells and in vitro induced islet-like cell transplantation can improve IR,lower blood glucose,and treat GDM.2.Compared with the hPDMSCs intervention group,the islet-like cell intervention group had a more significant decrease in blood glucose,and the relative expression of adiponectin mRNA was higher in the placental tissue,the relative expression of TNF-α and IL-6 mRNA was also more significant decrease.The therapeutic effect on GDM is better than that of uninduced hPDMSCs. |
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