2014Tropical Journal of Pharmaceutical ResearchOpen access

Anti-oxidative Effect of Ligustrazine on Treatment and Prevention of Atherosclerosis

GD Huang, Jiang‐Gao Mao, ZW Ji

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Abstract

Purpose: To investigate the protective effects of ligustrazine on oxidative stress-induced atherosclerosis.Methods: The indicators related to oxidative stress were determined using commercially available assay kits. MTT assay was used to assess the survival rate of human umbilical vein endothelial cells (HUVECs). HUVECs apoptosis was analyzed using fluorescence staining and flow cytometry. mRNA expression level and activity of caspases 3, 8, and 9 were determined via quantitative real-time polymerase chain reaction (PCR) and caspase 3, 8, and 9 assay kits.Results: Ligustrazine concentration of < 80 μmol/L had negligible inhibitory effect on HUVECs viability and protected HUVECs against oxygen stress damage by regulating the indicators related to oxidative stress. Flow cytometry results show that ligustrazine ameliorated H2O2-induced apoptosis, while the proportion of cells that stepped into early apoptosis and late apoptosis or necrosis were 52.7 and 0.6 %, respectively, in the H2O2 group, and 38.2 and 1.3 %, respectively, in the ligustrazine group. In addition, ligustrazine attenuated the up-regulation of caspase 3, 8, and 9 mRNA expression levels and activity.Conclusion: Ligustrazine can protect HUVECs against H2O2-induced injuries by regulating the indicators related to oxidative stress and suppressing the overexpression of caspases 3, 8, and 9. The protective mechanism of ligustrazine on H2O2-induced injury in HUVECs may be a caspase-dependent anti-apoptotic mechanism which provide important information for treating and preventing oxidative stress-induced atherosclerosis.Keywords: Ligustrazine, Oxidative stress, Umbilical vein, Endothelial cells, Atherosclerosis

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Purpose: To investigate the protective effects of ligustrazine on oxidative stress-induced atherosclerosis.Methods: The indicators related to oxidative stress were determined using commercially available assay kits. MTT assay was used to assess the survival rate of human umbilical vein endothelial cells (HUVECs). HUVECs apoptosis was analyzed using fluorescence staining and flow cytometry. mRNA expression level and activity of caspases 3, 8, and 9 were determined via quantitative real-time polymerase chain reaction (PCR) and caspase 3, 8, and 9 assay kits.Results: Ligustrazine concentration of < 80 μmol/L had negligible inhibitory effect on HUVECs viability and protected HUVECs against oxygen stress damage by regulating the indicators related to oxidative stress. Flow cytometry results show that ligustrazine ameliorated H2O2-induced apoptosis, while the proportion of cells that stepped into early apoptosis and late apoptosis or necrosis were 52.7 and 0.6 %, respectively, in the H2O2 group, and 38.2 and 1.3 %, respectively, in the ligustrazine group. In addition, ligustrazine attenuated the up-regulation of caspase 3, 8, and 9 mRNA expression levels and activity.Conclusion: Ligustrazine can protect HUVECs against H2O2-induced injuries by regulating the indicators related to oxidative stress and suppressing the overexpression of caspases 3, 8, and 9. The protective mechanism of ligustrazine on H2O2-induced injury in HUVECs may be a caspase-dependent anti-apoptotic mechanism which provide important information for treating and preventing oxidative stress-induced atherosclerosis.Keywords: Ligustrazine, Oxidative stress, Umbilical vein, Endothelial cells, Atherosclerosis

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Available abstract

Purpose: To investigate the protective effects of ligustrazine on oxidative stress-induced atherosclerosis.Methods: The indicators related to oxidative stress were determined using commercially available assay kits. MTT assay was used to assess the survival rate of human umbilical vein endothelial cells (HUVECs). HUVECs apoptosis was analyzed using fluorescence staining and flow cytometry. mRNA expression level and activity of caspases 3, 8, and 9 were determined via quantitative real-time polymerase chain reaction (PCR) and caspase 3, 8, and 9 assay kits.Results: Ligustrazine concentration of < 80 μmol/L had negligible inhibitory effect on HUVECs viability and protected HUVECs against oxygen stress damage by regulating the indicators related to oxidative stress. Flow cytometry results show that ligustrazine ameliorated H2O2-induced apoptosis, while the proportion of cells that stepped into early apoptosis and late apoptosis or necrosis were 52.7 and 0.6 %, respectively, in the H2O2 group, and 38.2 and 1.3 %, respectively, in the ligustrazine group. In addition, ligustrazine attenuated the up-regulation of caspase 3, 8, and 9 mRNA expression levels and activity.Conclusion: Ligustrazine can protect HUVECs against H2O2-induced injuries by regulating the indicators related to oxidative stress and suppressing the overexpression of caspases 3, 8, and 9. The protective mechanism of ligustrazine on H2O2-induced injury in HUVECs may be a caspase-dependent anti-apoptotic mechanism which provide important information for treating and preventing oxidative stress-induced atherosclerosis.Keywords: Ligustrazine, Oxidative stress, Umbilical vein, Endothelial cells, Atherosclerosis

Key concepts: Oxidative stress, Apoptosis, Umbilical vein, Flow cytometry, Chemistry, MTT assay, Pharmacology, Viability assay

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