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复方银杏颗粒对人脐静脉内皮细胞氧化应激损伤的保护作用及机制

[Protective effect and mechanism of compound Ginkgo biloba granules on oxidative stress injury of HUVEC].

作者信息

Li Qi, Chen Xi, Kan Xiao-Xi, Li Yu-Jie, Yang Qing, Wang Ya-Jie, Chen Ying, Weng Xiao-Gang, Cai Wei-Yan, Huang He-Fei, Zhu Xiao-Xin

机构信息

Institute of Chinese Materia Medica, China Academy of Chinese Medicinal Sciences, Beijing 100700, China.

School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2016 Feb;41(4):722-727. doi: 10.4268/cjcmm20160430.

Abstract

To reveal the protective and anti-apoptosis effect of compound Ginkgo biloba granules on oxidative stress injury of human umbilical vein endothelial cells (HUVEC). Negative control group, H2O2 model group and 4 drug pretreatment groups (80, 160, 320, 640 mg• L⁻¹) were established. The cell proliferation, morphological changes in each group after oxidative stress injury was detected by MTT assay and through microscope observation respectively. The content of LDH, MDA, SOD and NO and SOD activity in supernatant were detected to judge the protection effect of the drugs on endothelial cells. The protective effect on HUVEC apoptosis was analyzed by Caspase-3 activity test and Annexin V-FITC/PI staining. Western blot was used to observe the expression of apoptosis-related proteins Bcl-2 and Bax. Results showed that 1 200 μmol• L⁻¹ H2O2 can induce oxidative stress injury in endothelial cells and reduce the cell survival rate; cell proliferation inhibition degree is positively correlated with the effect time of H2O2. Besides, 80, 160, 320 640 mg•L⁻¹ compound Ginkgo biloba granules can protect HUVEC from oxidative stress injury, recover the normal proliferation level of cells, improve their state, prohibit cell apoptosis, and can up-regulate and down-regulate the expression level of Bcl-2 and Bax respectively. In conclusion, compound G. biloba granules can protect HUVEC from the oxidative stress injury induced by H2O2, its mechanism may be correlated with inhibition of the mitochondrial apoptotic pathway in HUVEC.

摘要

探讨复方银杏颗粒对人脐静脉内皮细胞(HUVEC)氧化应激损伤的保护及抗凋亡作用。设立阴性对照组、H2O2模型组和4个药物预处理组(80、160、320、640 mg•L⁻¹)。分别采用MTT法检测氧化应激损伤后各组细胞增殖情况,通过显微镜观察各组细胞形态变化。检测上清液中乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、一氧化氮(NO)含量及SOD活性,判断药物对内皮细胞的保护作用。通过Caspase-3活性检测及Annexin V-FITC/PI染色分析对HUVEC凋亡的保护作用。采用蛋白质免疫印迹法观察凋亡相关蛋白Bcl-2和Bax的表达。结果显示,1 200 μmol•L⁻¹ H2O2可诱导内皮细胞氧化应激损伤,降低细胞存活率;细胞增殖抑制程度与H2O2作用时间呈正相关。此外,80、160、320、640 mg•L⁻¹复方银杏颗粒可保护HUVEC免受氧化应激损伤,恢复细胞正常增殖水平,改善细胞状态,抑制细胞凋亡,并可分别上调和下调Bcl-2和Bax的表达水平。综上所述,复方银杏颗粒可保护HUVEC免受H2O2诱导的氧化应激损伤,其机制可能与抑制HUVEC线粒体凋亡途径有关。

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