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Aim To study the protective effects of Ganoderma lucidum polysaccharides peptide(GLPP)on ECV304 from oxidative injury.Methods Cultured ECV304 were injured by oxygen free radicals derived from tBOOH.Various concentrations of GLPP(12.5,25,50,100 mg?L-1)were added into culture medium.The survival rate of cells was measured by MTT assay.The morphological change of cells and injury of mitochondria were examined under the light and electron microscopes.The percentage of apoptosis of ECV304,labeled with AnnexinV/PI,was measured by flow cytometry.Results GLPP(12.5,25,50,100 mg?L-1)could reduce oxidative injury induced by tBOOH in ECV304 cells.The survival rate of cells treated with GLPP increased.The light microscopic examination showed that the injured cells decreased in GLPP-treated groups.Under the electron microscope it was found that GLPP(50 mg?L-1,incubated for 24 h)could protect the organelle such as mitochondria from oxidative injury and cells from apoptosis by tBOOH.The result of flow cytometry showed that the total percentage of apoptosis in control,GLPP and injury treated group was 2.24%?0.43%,24?6.4%(P
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Aim To study the effects of Ganoderma polysaccharide peptide (GLPP) on the production of nitric oxide (NO) in mice peritoneal macrophages induced by LPS and its mechanism. Methods The effects of GLPP on the NO releasing from mice peritoneal macrophages induced by LPS were detected by using Griess reagent and the expression of iNOS by GLPP in mice peritoneal macrophages was detected by immunohistochemical method.Result The production of NO was increased by GLPP.(25～200 mg?kg -1) ig for five days or by GLPP(3.125～200 mg?L -1) in vitro but the effects of LPS on the production of NO was not influenced significantly. The expression of iNOS was increased by GLPP(25～200 mg?kg -1) ig for five days or GLPP(3.125～200 mg?L -1) in vitro.Conclusion GPP in vivo or in vitro could increase the production of NO in mice peritoneal macrophages. It might take effects by enhancing the synthesis of iNOS.
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AIM: To investigate the free radical scavenging activity of Ganoderma lucidum polysaccharides peptide (GLPP) on peritoneal macrophages in mice. METHODS: Alloxan and tert butylhydroperoxide (tBOOH) were used as an oxidant to injury peritoneal macrophages in vivo in mice or in vitro, respectively. 2', 7' dichlorodihydrofluorescein diacetate (DCHF DA) was used as fluorescent probe. The fluorescence from cells was observed under the laser confocal microscope. Time series scan of conforcal microscope was used to observe the changes of fluorescence by GLPP in mice peritoneal macrophages over time. RESULTS: The results of confocal microscopy showed that GLPP (100 mg?kg -1 , ig for 5 d ) lowed fluorescence in the mice macrophages injured by alloxan (75 mg?kg -1 iv). GLPP (10 mg?L -1 ) also lowed fluorescence in the mice macrophages injured by tBOOH ( 7.76 ?10 -5 mol?L -1 ) in vitro as well. Time series scan showed that GLPP (10 mg?L -1 ) lowed fluorescence in the mice macrophages at rest state or during the respiratory burst induced by PMA (50 nmol?L -1 ). CONCLUSION: GLPP shows antioxidant effects and might have free radical scavenging effects on peritoneal macrophages in mice.
ABSTRACT
Aim To study the protective effects of Ganoderma lucidum polysaccharides peptide(GLPP)on the human umbilical vein endothelial cells(HUVECs)injured by reactive oxygen species(ROS),derived from tert-butylhydroperoxide(t-BOOH).Methods Vascular endothelial cells were obtained from infant umbilical cord for primary cultivation,identified by endothelial cell adhesion molecule-1(CD31)by fluorescence microscope.HUVECs were injured by ROS,derived from t-BOOH.The survival rate of cells was measured by Cell Counting Kit-8 assay.Hoechst 33258 nucleus staining was used to observe the early apoptosis injury by ROS.And the activation of Caspase-3 was detected by spectrophotometry.Electron microscopy was used to show the morphological changes in HUVECs.Results GLPP(6.125,12.5,25,50,100 mg?L-1)could inhibit the apoptosis of HUVECs by ROS.The survival rate of HUVECs was increased by GLPP and the percentage of cell apoptosis was decreased by it too.GLPP decreased the activation of Caspase-3 of HUVECs up-regulated by ROS.Electron microscope showed that the organelle such as mitochondria injury by ROS could be relieved by GLPP.Conclusion GLPP can prevent human umbilical vein endothelial cells from oxidative injury.