Involvement of CaSR in Hyperglycemia-induced Macroangiopathy and Related Mechanism  

作　　者：卢金萍 任江华 陈玲 李夏 陈慧莉 

机构地区：[1]Department of Internal Medicine & Geriatrics, Zhongnan Hospital of Wuhan University

出　　处：《Journal of Huazhong University of Science and Technology(Medical Sciences)》2015年第1期42-47,共6页华中科技大学学报（医学英德文版）

基　　金：supported by General Project of Health Department of Hubei Province,China(No.2013-JX6B75)

摘　　要：In order to clarify the potential role of calcium sensing receptor（Ca SR）, a typical G protein coupled receptor（GPCR）, in hyperglacemia-induced macroangiopathy, experimental hyperglycemia models in vivo and in vitro were prepared. Firstly, SD rats were divided into control group（n=10） and diabetes group（n=10）, and diabetic model was induced via high-fat diet feeding and streptozotocin（STZ, 30 mg/kg） injection. Hydroxyproline level, determined via Choramnie T oxidation method, in vessel wall in diabetic rats was 30% more than that in control group. The gene transcription and expression levels were detected by real-time PCR and Western blotting, respectively. Both of collagen Ⅰ and Ⅲ mR NA levels in diabetic aorta were nearly twice those in normal aorta. The cleaved caspase-3 and-9 were elevated 1.5 and 2.5 times respectively in diabetic vascular cells. As compared with controls, m RNA and protein levels of CaS R in aorta were increased by 3 and 1.5 times in diabetes group. The expression levels of Bax as well as pro-apoptotic kinases（phospho-p38 and phosphor-JNK） were also increased 2, 0.5 and 0.5 times respectively in diabetic rats. To further validate the involvement of Ca SR in cell apoptosis and explore the potential mechanism, the endothelial cell line（human umbilical vascular endothelial cells, HUVECs） was stimulated with high concentration of glucose（33 mmol/L） to mimic hyperglycemia in vitro. Cell-based assays also showed that the Ca SR level and key apoptotic proteins（cleaved caspase-3 and-9, Bax, phospho-p38 and phosphor-JNK） were elevated in response to stimulation, and inhibition of Ca SR by using specific inhibitor（NPS-2143, 10 μmol/L） could protect cells against apoptosis. Our results demonstrated that Ca SR might take important part in the development of diabetic macroangiopathy through promoting cell apoptosis induced by hyperglycemia.In order to clarify the potential role of calcium sensing receptor（Ca SR）, a typical G protein coupled receptor（GPCR）, in hyperglacemia-induced macroangiopathy, experimental hyperglycemia models in vivo and in vitro were prepared. Firstly, SD rats were divided into control group（n=10） and diabetes group（n=10）, and diabetic model was induced via high-fat diet feeding and streptozotocin（STZ, 30 mg/kg） injection. Hydroxyproline level, determined via Choramnie T oxidation method, in vessel wall in diabetic rats was 30% more than that in control group. The gene transcription and expression levels were detected by real-time PCR and Western blotting, respectively. Both of collagen Ⅰ and Ⅲ mR NA levels in diabetic aorta were nearly twice those in normal aorta. The cleaved caspase-3 and-9 were elevated 1.5 and 2.5 times respectively in diabetic vascular cells. As compared with controls, m RNA and protein levels of CaS R in aorta were increased by 3 and 1.5 times in diabetes group. The expression levels of Bax as well as pro-apoptotic kinases（phospho-p38 and phosphor-JNK） were also increased 2, 0.5 and 0.5 times respectively in diabetic rats. To further validate the involvement of Ca SR in cell apoptosis and explore the potential mechanism, the endothelial cell line（human umbilical vascular endothelial cells, HUVECs） was stimulated with high concentration of glucose（33 mmol/L） to mimic hyperglycemia in vitro. Cell-based assays also showed that the Ca SR level and key apoptotic proteins（cleaved caspase-3 and-9, Bax, phospho-p38 and phosphor-JNK） were elevated in response to stimulation, and inhibition of Ca SR by using specific inhibitor（NPS-2143, 10 μmol/L） could protect cells against apoptosis. Our results demonstrated that Ca SR might take important part in the development of diabetic macroangiopathy through promoting cell apoptosis induced by hyperglycemia.

关 键 词：calcium sensing receptor DIABETES HYPERGLYCEMIA MACROANGIOPATHY APOPTOSIS 

分 类 号：R543[医药卫生—心血管疾病]