白术内酯Ⅲ对2型糖尿病大鼠血管内皮细胞损伤、炎症、氧化应激的影响研究OA

Objective To investigate the effects of atractylenolide Ⅲ on vascular endothelial cell injury,inflammation,and oxidative stress in type 2 diabetes mellitus(T2DM)rats.Methods A T2DM rat model was established by feeding a high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin(STZ).After successful modeling,the rats were divided into the model group,atractylenolide Ⅲ low-dose group(gavaged with 25 mg/kg atractylenolide Ⅲ daily),atractylenolide Ⅲ high-dose group(gavaged with 50 mg/kg atractylenolide Ⅲ daily),atractylenolide Ⅲ high-dose+H-89[cyclic adenosine monophosphate(cAMP)inhibitor]group(gavaged with 50 mg/kg atractylenolide Ⅲ and intraperitoneally injected with 2 mg/kg H-89 daily),with a control group set up additionally;each group had 10 rats.After 4 weeks of intervention,rat body weight was measured,and fasting blood glucose(FBG)was detected using a glucometer.Serum levels of fasting insulin(FINS),interleukin(IL)-1 β,IL-10,tumor necrosis factor-α(TNF-α),and cAMP were measured by enzyme-linked immunosorbent assay.Plasma levels of nitric oxide(NO),endothelin-1(ET-1),and von Willebrand factor(vWF)were detected by double-antibody sandwich method.Superoxide dismutase(SOD)activity in thoracic aortic tissue was measured by hydroxylamine method,and malondialdehyde(MDA)content was measured by BAT method.Hematoxylin-eosin staining was used to observe the morphology of thoracic aortic tissue.The ultrastructure of rat thoracic aortic endothelial cells was observed under transmission electron microscopy.Protein expressions of proliferating cell nuclear antigen(Ki-67),cleaved caspase-3,protein kinase A(PKA),and cAMP response element-binding protein(CREB)in thoracic aortic tissue were detected by Western blotting.Results The control group showed no significant damage to thoracic aortic tissue and endothelial cell morphology.Compared with the control group,the model group exhibited severe damage to thoracic aortic tissue and endothelial cells,as well as increased levels of FBG,HOMA-IR,serum FINS,IL-1 β and TNF-α,plasma ET-1 and vWF,thoracic aortic tissue MDA content,and cleaved caspase-3 protein expression(P＜0.05),while body weight,serum IL-10 and cAMP levels,plasma NO level,thoracic aortic tissue SOD activity,and protein expression levels of Ki-67,phosphorylation-PKA/PKA,phosphorylation-CREB/CREB were decreased(all P＜0.05).Compared with the model group,the atractylenolide Ⅲ low-and high-dose groups showed significantly alleviated damage to thoracic aortic tissue and endothelial cells,as well as decreased levels of FBG,HOMA-IR,serum FINS,IL-1 β and TNF-α,plasma ET-1 and vWF,thoracic aortic tissue MDA content,and cleaved caspase-3 protein expression(all P＜0.05),and increased body weight,serum IL-10 and cAMP levels,plasma NO level,thoracic aortic tissue SOD activity,and protein expression levels of Ki-67,phosphorylation-PKA/PKA,phosphorylation-CREB/CREB(all P＜0.05).The cAMP inhibitor H-89 partially reversed the protective effects of atractylenolide Ⅲ on vascular endothelial cells in T2DM rats(P＜0.05).Conclusion Atractylenolide Ⅲ can reduce inflammatory response and oxidative stress in T2DM rats,thereby alleviating vascular endothelial cell injury,which may be related to the activation of the cAMP/PKA/CREB signaling pathway.