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白藜三醇对人脐静脉内皮细胞增殖及NO表达的影响
引用本文:史彤彤,;程明月,;张超群,;张卓琦,;王志荣.白藜三醇对人脐静脉内皮细胞增殖及NO表达的影响[J].徐州医学院学报,2014(7):477-480.
作者姓名:史彤彤  ;程明月  ;张超群  ;张卓琦  ;王志荣
作者单位:[1]徐州医学院研究生学院2011级,江苏徐州221002; [2]徐州医学院附属医院心内科,江苏徐州221004
基金项目:江苏省兴卫工程医学重点人才资助项目(RC2007089)
摘    要:目的观察白藜三醇对正常状态下人脐静脉内皮细胞(human umbilical vein endothelial cells,HU—VECs)磷酸化蛋白激酶B(P—Akt)、磷酸化内皮型一氧化氮合酶(P-NOS)及一氧化氮(NO)表达的调节作用,探讨白藜三醇对HUVECs增殖的影响及可能机制。方法实验分组:DMEM高糖(〈4.5mg/L)培养基培养组(正常组)、白藜三醇组(分为0.2、1、5、10、20μmol/L5个剂量亚组)、白藜三醇+LY294002组、LY294002组(阻断剂组)。其中LY294002为磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,P13-K)抑制剂。应用CCK-8法检测细胞增殖情况;Westernblot检测各组细胞内蛋白激酶B(Akt)、P—Akt、内皮型一氧化氮合酶(eNOS)、P-eNOS蛋白的表达;硝酸还原酶法检测细胞培养液中NO的浓度。结果①白藜三醇组1、5μmol/L2个剂量亚组与正常组相比细胞增殖增加(P〈0.01),5μmol/L亚组与0.2、10、20μmol/L亚组相比细胞增殖增加(P〈0.01);白藜三醇20μmol/L对细胞增殖有抑制作用(P〈0.01)。②与对照组比较,白藜三醇组P-Akt(P〈0.01)、P-eNOS(P〈0.05)蛋白表达增加;与白藜三醇组比较,白藜三醇+LY294002组P-Akt、P-eNOS蛋白表达降低(P〈0.01)。③与对照组比较,白藜三醇组NO表达增加(P〈0.05);与白藜三醇组比较,白藜三醇+LY294002组NO表达降低(P〈0.01)。结论白藜三醇可能通过P13-K/Akt/eNOS信号通路诱导正常状态下内皮细胞NO表达的增加,从而促进内皮细胞增殖。

关 键 词:白藜三醇  人脐静脉内皮细胞  一氧化氮  增殖

Effects of resveratrol on the proliferation of HUVECs and expression of NO
Institution:SHI Tongtong, CHENG Mingyue, ZHANG Chaoqun, ZHANG Zhuoqi, WANG Zhirong (1. Grade 2011, Graduate School, Xuzhou Medical College, Xuzhou, Jiangsu 221002, China; 2. Department of Cardiology, the Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangsu 221004)
Abstract:Objective To investigate the regulating effects of resveratrol on the expressions of phospho - Akt ( p - Akt), phospho- eNOS (p -eNOS) and nitric oxide (NO) in human umbilical vein endothelial cells (HUVECs) . The effects of resveratrol on the proliferation of HUVECs and its possible mechanisms were also investigated. Methods Grouping: high glucose ( 〈4.5 mg/L) DMEM medium group (normal group), resveratrol group (including 0.2, 1,5, 10, 20μmol/L subgroups) , resveratrol -LY294002 group, LY294002 group (blocker group). LY294002 is the bloc- ker of phosphatidylinositol 3 - kinase ( PI3 - K). CCK8 assay was used to detect the effects of reaveratrol on the prolifer- ation of HUVECs. Western blot was used to detect the p - Akt and the p - eNOS protein level respectively. NO concen- trations in conditioned media were determined using the nitrate reductase methods. Results (1) CCK8 : resveratrol group (1, 5 Ixmol/L) increased cell proliferation compared with normal group (P 〈 0.01 ) ; resveratrol (5μmol/L) increased cell proliferation compared with the drug group (0.2, 10, 20 μmol/L) (P 〈 0.01 ) ; resveratrol (20μmol/L) decreased cell proliferation compared with the normal group (P 〈 0.01 ). (2) Western blot: p- Akt (P 〈 0.01 ), p- eNOS (P 〈 0.05) expressions were increased significantly in resveratrol group compared with normal group; and p -Akt (P 〈 0.01 ) , p - eNOS (P 〈0. 01 ) expressions were decreased in resveratrol - blocker group compared with resveratrol group. (3) Nitric oxide assays: the culture media NO content of HUVECs was increased significantly in resveratrol group compared with normal group ( P 〈 0.05 ) , and it was decreased in resveratrol - blocker group compared with resveratrol group (P 〈 0.01 ). Conclusion Resveratrol can increase of expression of NO in HUVECs via PI3 - K/Akt/eNOS pathway, and further promote the proliferation of HUVECs.
Keywords:resveratrol  human umbilical vein endothelial cells  nitric oxide  proliferation
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