Effects Of High Glucose And Insulin On MSCs Proliferation And The Possible Mechanism

ObjectiveTo investigate effects of high glucose and insulin on MSCs proliferation and then detect cell cycle and apoptosis for possible mechanisms.MethodsBone marrow cells were obtained centrifugally from femora of 8-week-aged mice and MSCs isolated by the method of percoll centrifugation ,then After the cells reached 70%-80% confluence, evaluated the purity coefficient of MSCs by cell cycle detection and morphology . established different groups of glucose concentrations(5.6mmol/L,11mmol/L and 25mmol/L), and below 11mmol/L and 25mmol/L glucose, we set four insulin concentration groups(0μU/mL I,15μU/mL ,30μU/mL and 200μU/mL). After 72h intervention, detected indexs as below: (1)proliferation : by MTT assay.(2) cell cycle: analyzed for cell cycle distribution by a flow cytometry system.(3) apoptosis : Annexin V/PI analysis. All data were analyzed by SPSS software system. Experimental groups were compared by ANOVA, and when appropriate, with LSD analysis for multiple comparisons.ResultsMSCs was obtained and the purity coefficient of them exceeded 90%. Single glucose affected MSCs proliferation in two-way(P=0.003), that is , middle glucose(11mmol/L) increased the rate of MSCs proliferation remarkably, but its exorbitant concentration(25mmol/L) let this effect disappear respectively;High glucose increased the percent of S phase in cell cycle(P=0.002)and the rate of apoptosis(P=0.000)significantly. Within the groups of MHG and EHG, Insulin also affected MSCs proliferation in two-way(P=0.000,P=0.004), that is , physiogical Insulin(15mmol/L) increased the rate of MSCs proliferation remarkably, but increased concentration let this effect disappear ;Insulin had no effects on cell cycle(P=0.066, P=0.840), but decreased the rate of apoptosis caused by high glucose obviously(P=0.021,P=0.001).Conclusions1. Both glucose and insulin have created a biphasic effect toward MSCs proliferation, and there is cooperation between them.2. Cell cycle and apoptosis are responsible for the alteration of MSCs proliferation partly. Maybe there are other coexistent mechanisms .