| Objective:Study on the influence of abnormal savda munziq is rich in polyphenols extract on the proliferation of PC 12 cells and prevent the protection effect of PC 12 cell H2O2 oxidative damage, and to explore its mechanism of action.Methods:Combination of polyamide resin adsorption ASM-PE was prepared by alcohol, expressed as gallic acid, Folin phenol (Folin-Ciocalteu, FC) determination of ASM ethanol extract and purify total phenolic content in separation and colorimetric method; through the influence of detection of ASM-PE MTT colorimetric method on the proliferation of PC 12 cells. At the same time induced by H2O2 to establish the oxidative damage model of PC 12 cells, the effects by MTT method and morphological detection of ASM-PE on H2O2 induced PC 12 cell damage and cell survival rate, further to determine the protective effect of ASM-PE PC 12 cells from H2O2 damage; prevent the changes in cell morphology characteristics and by Annexin-V/Pl double staining flow cytometry ASM-PE influence on PC 12 cell apoptosis by Hoechst 33342 staining for detection of ASM-PE. In the study of the protective mechanism of ASM-PE in detecting the activity of antioxidant enzymes, liquid SOD in PC 12 cell culture, the discussion between ASM-PE subjected to oxidative stress in PC12 cells oxidative and antioxidative system balance influence; Then by immunofluorescence detection and Western blot of ASM-PE protect PC12 cells against oxidative damage to H2O2, expression of related cell factor Caspase-3 and Bcl-2 cell apoptosis, thus a preliminary discussion on the mechanism of ASM-PE exerts a protective effect.Results:(1) By extraction and purification steps, the ASM-PE content in the extracts was 26.58%;(2) In a certain range of concentration, the detection results found that ASM-PE was cytotoxic effect on PC 12 cells; (3) Detected by MTT method and morphological method that ASM-PE can significantly reduce the oxidative damage resulting from PC 12 cells induced by H2O2.(4) By morphological studies that ASM-PE can enhance PC 12 cell induced by damage tolerance for H2O2, reduce the occurrence of nuclear condensation and marginalization and apoptotic bodies appeared as apoptosis of characterization of the phenomenon,’can reduce the apoptosis of PC 12 cells induced by H2O2; (5) By apoptosis detection of Hoechst 33342 staining that ASM-PE to apoptosis PC 12 cells was significantly less than the injury group, weaken the oxidative H2O2 damage in PC 12 cells; (6) Through the cell supernatant SOD levels determined that ASM-PE can significantly inhibit the antagonism of H2O2 on PCI2 cell activity of SOD; (7) The amount of expression of ASM-PE can inhibit the enzyme prototype of Caspace-3 apoptosis protein by immunofluorescence and Western blot detection of apoptosis related gene expression, and enhance the expression of anti apoptosis protein Bcl-2. Conclusion: This study concluded that, in a certain range of concentration, ASM-PE was not toxic to PC 12 cells, and can be attenuated H2O2 induced oxidative damage occurs in PC 12 cells, apoptosis occurs to improve the characterization of phenomena. The possible mechanisms for ASM-PE:by increasing the antioxidant enzymes such as SOD, decrease due to H2O2 induced ROS; in addition, the mechanism of ASM-PE play a protective role may also trigger factor by inhibiting apoptosis (such as ROS) to activate the expression of Caspase-3, apoptosis promoting factor, and enhanced Bcl-2 anti apoptosis factor expression, regulation of cell apoptosis signal pathway, play a protective cell survival effect. |
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