野蔷薇根总黄酮对H_(2)O_(2)诱导损伤的HUVEC线粒体功能及凋亡通路的影响  

Effect of Total Flavonoids from Rosa multiflora Thunb.on Mitochondrial Function and Apoptin Pathway of HUVEC Induced by Hydrogen Peroxide

作　　者：李开言[1] 王军[1] 杨丹 张留记[1] 屠万倩[1] 吴冰帆 LI Kaiyan;WANG Jun;YANG Dan;ZHANG Liuji;TU Wanqian;WU Bingfan(Henan Provincial Academy of Traditional Chinese Medicine,Zhengzhou 450004 Henan,China;Henan University of Traditional Chinese Medicine,Zhengzhou 450008 Henan,China)

机构地区：[1]河南省中医药研究院,河南郑州450004 [2]河南中医药大学,河南郑州450008

出　　处：《中药新药与临床药理》2021年第10期1431-1436,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：河南省中医药科学研究专项课题(2019ZY2013)。

摘　　要：目的探讨野蔷薇根总黄酮(TF-RM)对H_(2)O_(2)诱导损伤的人脐静脉内皮细胞(HUVEC)线粒体功能及凋亡通路的影响。方法将SD雄性大鼠随机分正常对照组、维生素C组(0.08 g·kg^(-1))、TF-RM组(2.5 g·kg^(-1)),每组10只。灌胃给药,每天1次,连续7 d,制备含药血清。采用H_(2)O_(2)(950μmol·L^(-1))诱导HUVEC损伤,并分别与维生素C及TF-RM低、中、高剂量(5%、10%、15%)含药血清共培养24 h。采用DCFH-DA荧光定量法检测HUVEC中活性氧(ROS)的含量;JC-1染色法检测细胞线粒体膜电位的变化;Fluo-4染色法检测细胞内Ca^(2+)浓度;Western Blot法检测B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、半胱氨酸天冬氨酸特异性蛋白酶(Caspase)-9、Caspase-3蛋白的表达情况。结果与正常对照组比较,模型组HUVEC内的ROS水平、线粒体膜电位下降率、Ca^(2+)浓度以及Bax蛋白表达量显著升高,差异均有统计学意义(P<0.01)。与模型组比较,TF-RM各剂量组细胞内的ROS水平、Ca^(2+)浓度及Bax蛋白表达量均明显降低(P<0.05,P<0.01),Bcl-2蛋白表达量升高(P<0.05);TF-RM高剂量组的线粒体膜电位下降率明显降低(P<0.05);TF-RM中剂量组的Caspase 9蛋白表达量明显降低(P<0.05)。结论TF-RM能够改善H_(2)O_(2)诱导的HUVEC线粒体功能损伤,发挥对细胞凋亡的保护作用。Objective To study the effect of total flavonoids from Rosa multiflora Thunb.(TF-RM)on mitochondrial function and apptosis pathway of HUVEC induced by hydrogen peroxide.Methods Male SD rats were randomly divided into normal control group,Vitamin C group(0.08 g·kg^(-1))and TF-RM group(2.5 g·kg^(-1)),10 in each group.After intragastric administration once a day for seven consecutive days,rats were anesthetized to prepare drugcontaining serum.Hydrogen peroxide(950μmol·L^(-1))was used to induce damage in HUVEC;and Vitamin C,serum containing TF-RM of high,medium and low dosages(5%,10%,15%)were added to HUVEC respectively at the same time.Then intracellular reactive oxygen species(ROS)was detected by DCFH-DA fluorescence quantitative assay;mitochondrial membrane potential(JC-1)was detected by JC-1 staining;decrease rate of Ca^(2+)concentration was detected by Fluo-4 staining.The expressions of B-cell lymphoma-2(Bcl-2),Bcl-2 associated X Protein(Bax),Caspase-9 and Caspase-3 were detected by Western Blot.Results Compared with normal control group,the ROS,decrease rate of mitochondrial membrane potential,Ca^(2+)concentration and the expression of Bax were improved when treated with hydrogen peroxide,the differences were statistically significant(P<0.01).Compared with the model group,the ROS,Ca^(2+)concentration and the expression of Bax were decreased,the differences were statistically significant(P<0.05 or P<0.01)while the expression of Bcl-2 was increased,the differences were statistically significant(P<0.05)when treated with TF-RM low,medium and high dosage,respectively.The decrease rate of mitochondrial membrane potential was decreased when treated with TF-RM high dosage,the difference was statistically significant(P<0.05).The expression of Caspase 9 was decreased when treated with TF-RM medium dosage,the difference was statistically significant(P<0.05).Conclusion TF-RM could improve the H_(2)O_(2)-induced mitochondrial function damage of HUVEC and play a protective role in apoptosis.

关 键 词：野蔷薇根总黄酮 人脐静脉内皮细胞 凋亡 线粒体功能 H2O2 

分 类 号：R285.5[医药卫生—中药学]