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[山核桃叶总黄酮对缺氧/复氧损伤培养的H9C2心肌细胞的保护作用]

[Protective Effect of Total Flavonoids from Carya cathayensis Leaf on Cultured H9C2 Cardiomyocytes During Hypoxia / Reoxygenation Injury].

作者信息

Jiang Rui-bin, Zhu Xue-xin, Xing Li-wan, Ding Zhi-shan, Lv Gui-yuan, Jin Bo

出版信息

Zhong Yao Cai. 2016 Feb;39(2):378-82.

PMID:30080372
Abstract

OBJECTIVE

To study the protective effect of total flavonoids from Carya cathayensis leaf on cultured H9C2 cardimyocytes during hypoxia / reoxygenation( H / R) injury.

METHODS

Co Cl2 was used to induce the H / R injury model in H9C2 cells at different concentrations,different H / R time. Total flavonoids from Carya cathayensis leaf was added into culture medium with final concentration of2. 5,5 and 10 μg / m L before H / R. Lactate dehydrogenase( LDH), Malondialdehyde( MDA) content, Superoxidedimutase( SOD) activity in the bathing medium were assayed for the evaluation of myocardial cell injury. Myocardial cell viability was detected by the MTS assay kit. Apoptotic changes in H9C2 cells were observed by using Hoechst-PI staining and flow cytometry analysis. The expression of HIF-1αprotein was detected by Western blot.

RESULTS

Using 1 200 μmol / L Co Cl2 to hypoxia the cells for 18 h and then reoxygenation for 2 h were the best condition of H / R injury model. Compared with model group,the cells treated with total flavonoids from Carya cathayensis leaf could decrease the activity of LDH and the content of MDA, while increased the activity of SOD. Moreover, they could regulate the expression of HIF-1α protein to normal level.

CONCLUSION

It is suggested that total flavonoids from Carya cathayensis leaf has a protective effect on H9C2 cardiomyocyte during H / R injury. The mechanism may be relate to enhancing the capability of the cell clearing the oxygen free radial, decreasing the production of lipid peroxidation and reducing apoptosis.

摘要

目的

研究山核桃叶总黄酮对缺氧/复氧(H/R)损伤的培养H9C2心肌细胞的保护作用。

方法

采用不同浓度、不同H/R时间的CoCl₂诱导H9C2细胞建立H/R损伤模型。在H/R之前,将山核桃叶总黄酮加入培养基中,终浓度分别为2.5、5和10μg/mL。检测培养液中乳酸脱氢酶(LDH)、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性,以评估心肌细胞损伤情况。采用MTS检测试剂盒检测心肌细胞活力。通过Hoechst-PI染色和流式细胞术分析观察H9C2细胞的凋亡变化。采用蛋白质免疫印迹法检测HIF-1α蛋白的表达。

结果

用1200μmol/L CoCl₂使细胞缺氧18 h,然后复氧2 h是建立H/R损伤模型的最佳条件。与模型组相比,山核桃叶总黄酮处理的细胞可降低LDH活性和MDA含量,同时提高SOD活性。此外,还可将HIF-1α蛋白的表达调节至正常水平。

结论

山核桃叶总黄酮对H/R损伤的H9C2心肌细胞具有保护作用。其机制可能与增强细胞清除氧自由基的能力、减少脂质过氧化产物的生成及减少细胞凋亡有关。

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