当归甙调控FOXO1干预高血压模型大鼠的血管内皮功能障碍  

Sweroside intervenes with vascular endothelial dysfunction in hypertensive rats by regulating FOXO1 expression

作　　者：张希倩 谭高峰[1] 孙晓泽[1] 庞欣[2] 韩宇 Zhang Xiqian;Tan Gaofeng;Sun Xiaoze;Pang Xin;Han Yu(Department of Geriatrics,Henan Provincial Hospital of Traditional Chinese Medicine,Zhengzhou 450000,Henan Province,China;Department of Nephrology,Henan Provincial Hospital of Traditional Chinese Medicine,Zhengzhou 450000,Henan Province,China;Department of Gastroenterology,Henan Provincial Hospital of Traditional Chinese Medicine,Zhengzhou 450000,Henan Province,China)

机构地区：[1]河南省中医院老年病科,河南省郑州市450000 [2]河南省中医院肾病科,河南省郑州市450000 [3]河南省中医院消化内科,河南省郑州市450000

出　　处：《中国组织工程研究》2023年第23期3628-3634,共7页Chinese Journal of Tissue Engineering Research

基　　金：河南省中医药科学研究专项(20-21ZY2015),项目负责人:韩宇。

摘　　要：背景:当归甙可调控细胞氧化应激水平,但其在高血压诱导的内皮细胞氧化损伤中的作用尚不清楚。目的:探讨当归甙是否通过调控叉头框转录因子O1(Forkhead box O1,FOXO1)表达调控高血压诱导的血管内皮功能障碍。方法:(1)体内使用肾主动脉缩窄法建立高血压大鼠模型,40只雄性SD大鼠随机分为4组:假手术组;高血压组;高血压+25 mg/kg当归甙组;高血压+50 mg/kg当归甙组。采用TUNEL染色检测大鼠胸主动脉内皮细胞凋亡情况。(2)体外利用50μmol/L H_(2)O_(2)刺激人脐静脉血管内皮细胞建立细胞氧化损伤模型,分为:对照组;H_(2)O_(2)组;H_(2)O_(2)+当归甙组(10,20,40μmol/L当归甙);H_(2)O_(2)+Vector(阴性对照载体)组;H_(2)O_(2)+pcDNA-FOXO1组;H_(2)O_(2)+当归甙(40μmol/L)+Scramble(阴性对照载体)组;H_(2)O_(2)+当归甙(40μmol/L)+si-FOXO1组。采用MTT和流式细胞术检测细胞增殖和凋亡情况;采用ELISA检测血清和人脐静脉血管内皮细胞中血管紧张素Ⅱ、丙二醛和一氧化氮水平,以及胸主动脉组织和人脐静脉血管内皮细胞中血管细胞黏附分子1、细胞间黏附分子1和E-选择素水平;采用RT-qPCR和Western blot检测胸主动脉组织和人脐静脉血管内皮细胞中FOXO1 mRNA和蛋白表达。结果与结论:(1)在体内,与假手术组比较,高血压模型大鼠血压和细胞凋亡水平明显升高(P<0.05),FOXO1 mRNA和蛋白水平明显降低(P<0.05),血管紧张素Ⅱ、丙二醛、血管细胞黏附分子1、细胞间黏附分子1和E-选择素水平明显升高(P<0.05),一氧化氮水平明显降低(P<0.05)。当归甙以剂量依赖方式改善高血压大鼠血管功能障碍。(2)在体外,与对照组比较,H_(2)O_(2)组细胞增殖、FOXO1表达水平、一氧化氮水平明显降低(P<0.05),细胞凋亡及丙二醛、血管细胞黏附分子1、细胞间黏附分子1和E-选择素水平明显升高(P<0.05)。当归甙或FOXO1过表达可改善H_(2)O_(2)诱导的人脐静脉血管内皮细胞功能�BACKGROUND: Sweroside can regulate cellular oxidative stress, but its role in hypertension-induced oxidative damage of endothelial cells remains unclear.OBJECTIVE: To investigate whether sweroside regulates hypertension-induced vascular endothelial dysfunction by regulating Forkhead box O1(FOXO1)expression.METHODS:(1) In vivo: The renal aorta constriction method was used to establish a hypertensive rat model. Forty male Sprague-Dawley rats were randomly divided into four groups: sham group, hypertension group, hypertension+25 mg/kg sweroside group, and hypertension+50 mg/kg sweroside group. TUNEL staining was used to detect the apoptosis of rat thoracic aorta endothelial cells.(2) In vitro: 50 μmol/L H_(2)O_(2)was used to stimulate human umbilical vein endothelial cells to establish a cell oxidative damage model. Damaged cells were divided into control group, H_(2)O_(2)group, H_(2)O_(2)+10, 20, or 40 μmol/L sweroside groups, H_(2)O_(2)+Vector group(negative control), H_(2)O_(2)+pcDNA-FOXO1 group, H_(2)O_(2)+40 μmol/L sweroside+Scramble group(negative control), and H_(2)O_(2)+40 μmol/L sweroside+si-FOXO1 group. MTT and flow cytometry were used to detect cell proliferation and apoptosis, respectively. ELISA was used to detect the levels of angiotensin Ⅱ, malondialdehyde, and nitric oxide levels in rat serum and human umbilical vein endothelial cells, as well as vascular cell adhesion molecule 1,intercellular adhesion molecule 1, and E-selection levels in rat thoracic aorta tissues and human umbilical vein endothelial cells. RT-qPCR and western blot were used to detect the mRNA and protein expression of FOXO1 in rat thoracic aorta tissues and human umbilical vein endothelial cells.RESULTS AND CONCLUSION:(1) In vivo: compared with the sham group, blood pressure and apoptosis were significantly increased(P < 0.05), the expression levels of FOXO1 mRNA and protein were significantly reduced(P < 0.05), the contents of angiotensin Ⅱ, malondialdehyde, vascular cell adhesion molecule 1,intercellular adhesion molecul

关 键 词：当归甙 FOXO1 高血压 氧化应激 内皮细胞功能障碍 

分 类 号：R446[医药卫生—诊断学]