MiR-98 Inhibits The Vascular Endothelial Cell Proliferation Of Diabetic Rats By Targeting Cyclin D2

This thesis is divided into two main parts:Part 1, The role of miR-98 in regulating vascular endothelial cell proliferation of type 2 diabetic SD rats; Part 2, miR-98 inhibits RAOEC proliferation by regulating Cyclin D2. Part ?. The role of miR-98 in regulating vascular endothelial cell proliferation of type 2 diabetic SD ratsFirstly, type 2 diabetic rat model was established. Then, we detected the expression of miR-98 in rat vascular tissue by Real-time PCR.The experiment results showed that the expression of miR-98 in diabetic rat vascular tissue was lower than that in the tissue of health control rats, which indicates that miR-98 plays important role in the diabetic vascular endothelial cell proliferation. Then, we detected the expression of Cyclin D2 by western blotting, which showed that the expression of Cyclin D2 in diabetic rat vascular tissue was significantly higher than that of control tissue. In addition, the expression of p-Rb was also higher in the diabetic rat vascular tissue. In conclusion, cyclin D2 and its related phosphorylation of Rb, play an important role in vascular endothelial cell proliferation.In vitro, RAOEC was treated by high glucose, we detected the cell proliferation of RAOEC by MTT, which showed that the cell proliferation rate of experimental groups was higher than normal control. Then we detected the cell cycle after treated by high glucose. The cell proliferation rate was higher than normal control.Our results further showed that the expression of miR-98 was much lower in high glucose-treated cells than that in control cells. The expression of Cyclin D2 and p-Rb in high glucose group was obviously higher than that of healthy control tissues. In conclusion, cyclin D2 and its related phosphorylation of Rb, play an important role in vascular endothelial cell proliferation. Part ?. miR-98 inhibits RAOEC proliferation by regulating Cyclin D2.We structured the evertor pcDNA-GFP-cyclin D2-3'UTR and synthesized miR-98 and ASO-98. RAOEC was co-transfected with miR-98 and pcDNA-GFP-cyclin D2-3'UTR. The expression of GFP was detected by fluorescence microscope and FCM. Our results showed that the expression of GFP was significantly suppressed in miR-98-treated cells than that of controls. The cell proliferation inhibition ratio was higher in miR-98-transfected cells than that of controls by MTT analysis. Then we found that miR-98 can markedly promote the apoptosis of vascular endothelial cells, inhibit the proliferation of vascular endothelial cell in G1 cell cycle block.In vitro study, after RAOEC was transfected with miR-98, the expression of miR-98 was higher in miR-98-treated cells than that in control cultures by Real-time PCR. After transfected miR-98, the expression of Cyclin D2 and p-Rb were lower in miR-98-transfected cells than those in control treatment group by western blotting. In addition, The expression of Bcl-2 and NF-KB were higher, while the expression of BAX and Caspase9 were lower in miR-98-transfected cells compared with control treatment. These results that the suppressive roles of miR-98 in cell proliferation are related to the expression of these genes.In summary, Cyclin D2 plays an important role in type 2 diabetic rat vascular endothelial cell proliferation. miR-98 can inhibit proliferation of vascular endothelial cells by regulating the expression of Cyclin D2. miR-98 also affects the proliferation of vascular endothelial cells by suppressing Bcl-2?NF-KB?BAX and Caspase9.