The Mechanism Of Velvet Cold Extract Regulate Cell Growth And Apoptosis In Hypoxic Model

Objective: This study the establishment of cobalt chloride-induced rat pheochromocytoma PC12 cell tumor hypoxia model, by detecting velvet cold extract to normal training and hypoxia and hypoxia PC12 cell activity and apoptosis-related gene expression in, Cold extract of velvet on the protection of neurons in the role of hypoxia cellular and molecular mechanism. For the clinical application of velvet cold extract the treatment of ischemic disease hypoxic provide theoretical and experimental basis.Method: PC12 cells in culture, by adding the final concentration of 125 mmol / L of cobalt chloride, the establishment of Cocl2 mediated PC12 cells hypoxic damage model; MTT detected velvet extract on PC12 cell proliferation activity of cell morphology Observation of cell differentiation; flow cytometry of the cell cycle and apoptosis; RT-PCR detected gene bax, bcl-2, HIF1, GADD45 mRNA expression.Results:1. Successful application of this study to establish the PC12 cells Cocl2 hypoxia model. MTT was detected velvet cold extract of the PC12 cell proliferation hypoxia before and after the impact, the experiment shows that, compared with hypoxia, the velvet cold extract of the role of hypoxia significantly enhance the ability of cell proliferation. With statistical significance (P <0.05)2. Application flow cytometry test velvet cold extract of hypoxic damage model of cell cycle and apoptosis. The results showed that the G0/G1 phase of the control group for 55.83%, S-phase cells to 20.3 percent, to 0.40 percent rate of apoptosis, cell hypoxic injury group for the G0/G1 phase of 73.77%, S-phase cells to 9.15 percent, Diao Death rate was 10.60 percent; velvet cold extract role for the group G0/G1 phase of 67.79%, S-phase cells to 19.82 percent, 5.40 percent rate of apoptosis.3. RT-PCR method of detection PC12 model by hypoxia velvet extract role before and after the Cold bcl-2 and bax expression changes. The results showed that the extract of velvet cold, hypoxic injury of the bcl-2 expression increased bax expression decreased.4. By RT-PCR method to detect through the cold of velvet, the PC12 cells hypoxia model HIF1 and GADD45 expression. The results showed that, compared with hypoxia, velvet cold after extract, HIF1 gene expression markedly improved; GADD45 expression lower.5. Extract of the velvet cold after, PC12 cells from the former role of the radio, to differentiate into spindle that velvet cold extract can promote the differentiation of PC12 cells.Conclusion:1. Velvet cold extract can promote cell proliferation.2. Velvet cold extract can promote the proliferation of hypoxic injury.3. Velvet cold extract can promote nerve cell differentiation.4. Velvet cold to the role of hypoxic damage cells can inhibit apoptosis; can restrain apoptosis gene expression of bcl-2mRNA increased, and will promote apoptosis gene expression baxmRNA weakened. Velvet cold extract of hypoxia can inhibit apoptosis.5. Velvet cold extract can increase HIF1 expression of hypoxia, decreased GADD45 expression, improve the ability of hypoxic cells resistant to oxygen.