舒洛地特激活AMPK/SIRT1通路减轻高糖诱导的大鼠心肌微血管内皮细胞损伤  

Sulodexide alleviates high glucose-induced cardiac microvascular endothelial cell injury in rat by activating the AMPK/SIRT1 pathway

作　　者：高砚丽 任引刚 汪杰 康黎 GAO Yanli;REN Yingang;WANG Jie;KANG Li(Department of Geriatric Medicine,Tangdu Hospital of Air Force Military Medical University,Xi’an 710038,China)

机构地区：[1]空军军医大学第二附属医院老年医学科,西安710038

出　　处：《广西医科大学学报》2024年第7期1003-1008,共6页Journal of Guangxi Medical University

基　　金：空军军医大学第二附属医院社会人才资助项目(No.2021SHRC025)。

摘　　要：目的:探讨舒洛地特(SDX)减轻高糖(HG)诱导的大鼠心肌微血管内皮细胞(CMEC)氧化应激和凋亡的作用及其机制。方法:分离培养大鼠CMEC,随机分为对照组、HG组、HG+SDX组。CCK-8法检测细胞存活率。利用DCFH-DA作为荧光探针,测定各组DCF荧光强度以判定细胞内活性氧(ROS)水平。比较各组细胞上清液中超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。采用流式细胞术和TUNEL法检测细胞凋亡率,western blotting法检测AMPK、磷酸化(p-)AMPK、SIRT1蛋白表达。结果:与对照组相比,HG组CMEC细胞存活率降低,细胞上清液中MDA含量增高,SOD活性降低,ROS水平和细胞凋亡率升高,p-AMPK、SIRT1蛋白表达水平下降(均P<0.05)。与HG组比较,HG+SDX组CMEC的细胞存活率显著提升,MDA、ROS生成减少,SOD活性升高,CMEC凋亡率下降,p-AMPK、SIRT1蛋白表达水平升高(均P<0.05)。结论:SDX可能通过激活AMPK/SIRT1信号通路,减轻HG诱导的CMEC的氧化应激及细胞凋亡,从而发挥心血管保护作用。Objective:To study the effect and mechanism of sulodexide(SDX)on the oxidative stress and apop-tosis of cardiac microvascular endothelial cells(CMEC)in rats under high glucose(HG).Methods:CMEC of rats were isolated and cultured,and randomly divided into control group,HG group and HG+SDX group.Cell survival rates were measured by cell counting kit-8(CCK-8)assay.DCFH-DA was used as a fluorescence probe to determine the fluorescence intensity of DCF in each group to determine the level of intracellular reactive oxy-gen species(ROS).The activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)in the supernatant were compared.The apoptosis rate was detected by flow cytometry and TUNEL assay.The ex-pression of AMPK,phosphorylated(p-)AMPK and SIRT1 proteins was detected by western blotting.Results:Compared with the control group,the HG group showed decreased cell survival rate,increased cell supernatant level of MDA,reduced SOD activity,elevated ROS level,increased apoptosis rate,and decreased expression of p-AMPK、SIRT1 proteins(all P<0.05).Compared with the HG group,the survival rate of CMEC cells in the HG+SDX was significantly increased,the production of MDA and ROS was decreased,the activity of SOD was in-creased,the apoptosis rate of CMEC was decreased,and the expression of p-AMPK and SIRT1 proteins was sig-nificantly increased(all P<0.05).Conclusion:SDX may alleviate HG-induced oxidative stress and apoptosis of CMEC by activating AMPK/SIRT1 signaling pathway,thus playing a cardiovascular protective role.

关 键 词：舒洛地特 高糖 心肌微血管内皮细胞 氧化应激 凋亡 

分 类 号：R542.2[医药卫生—心血管疾病]