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Alzheimer's disease (Alzheimer's disease, AD) is a progressive memory loss, cognitive impairment, and personality changes as the main clinical characteristics of the central nervous system degenerative diseases, has become the following after heart disease, cancer and stroke No. 4 killer, its main pathological features of brain senile plaque deposition, neurofibrillary tangles and neuronal degeneration. AD has a complex pathophysiological process, affected by many factors, including oxidative stress plays an important role in the pathogenesis of AD process. Lentinan (lentinan, LNT) is extracted from mushrooms daughter, separated, purified polysaccharide fractions, improve immune, anti-tumor, anti-viral, liver detoxification, antioxidant, hypoglycemic, lipid-lowering, anti- inflammation of the role of anti-infection. The lentinan clinical adjuvant therapy for cancer, hepatitis, etc., but its role in AD remains to be further studied. The subject using hydrogen peroxide (hydrogen peroxide, H2O2) as an exogenous free radical generating system, SH-SY5Y cells instead of nerve cells as an experimental object analog neural cells from oxidative damage vitro nerve cells from oxidative stress injury model, oxidative damage observed lentinan protective effect of SH-SY5Y cells and its possible mechanism, to provide the basis for its treatment for AD. Lentinan H2O2 injury SH-SY5Y cell morphology cells treated after inversion under the microscope and photographed. The experimental results show that: the group of cells in normal cell body fusiform, irregular triangle or polygon, shorter axons and dendritic-like protrusions; the H2O2 model group reduced cell number, cell body shrinkage, and some cells were round; Compared with the model group, lentinan concentration protection group increased cell number, cell body shrinkage phenomenon significantly improved. Lentinan H2O2 injury in SH-SY5Y cell viability by MTT assay cell viability. The experimental results show that: the SH-SY5Y cells after H2O2 injury, cell viability was significantly lower (P lt; 0.01); compared with the model group, 0.1 mg / L lentinan protection group cell viability enhanced (P lt; 0.05), 1 10 mg / L lentinan protection group, cell viability was significantly enhanced (P lt; 0.01). Lentinan LDH leakage rate of H2O2 injury in SH-SY5Y cells collected SH-SY5Y cells in the treatment of oxidative damage to the culture supernatant and cell lysate, referring to the kit instructions were measured in the culture supernatant and cell lysate lactate dehydrogenase (lactate dehydrogenase, LDH) activity of cells LDH leakage rate. Experimental results show that: of H2O2 model group SH-SY5Y cell membrane permeability increased LDH leakage rate was significantly higher (P lt; 0.01); compared with the model group, given 0.1,1,10 mg / L lentinan, inhibit cell the release of LDH was significantly decreased cell LDH leakage rate (P lt; 0.01). 4 of Lentinan of H2O2 injury in SH-SY5Y cells MDA content and SOD collect cell lysates, the amount of total protein with Coomassie Brilliant Blue statutory intracellular reference malondialdehyde (malondialdehyde, MDA), superoxide dismutase (superoxidase dismutase, SOD) assay kit were measured intracellular content of MDA, SOD activity. Experimental results show that: MDA content in the cells by H202 oxidative damage after a significant increase (P lt; 0.01), SOD activity was significantly lower (P lt; 0.01); via lentinan 0.1,1,10 mg / L after treatment, the intracellular content of MDA a significant reduction (P lt; 0.01), SOD activity was significantly higher (P lt; 0.01). Lentinan H2O2 injury in SH-SY5Y nuclear morphology cells after treatment at room temperature to 4% paraformaldehyde fixed, Hoechst 33258 staining, observed and photographed under a fluorescence microscope. The experimental results show that: the cells after Hoechst 33258 staining, normal group uniform staining of cell nuclei; visible dense granular fluorescence the H2O2 model group within the cell nucleus, showing the performance of apoptotic cells typical nuclear pyknosis, nuclear fragmentation; lentinan The cell is the nucleus morphology of each concentration protection group tended to be normal, and significantly reduced the number of apoptotic cells. 6. The Lentinan of H2O2 injury in SH-SY5Y cell apoptosis rate cells treated to on the the Annexin V-FITC/PI after transfection by flow cytometry. The experimental results show that: Compared with the normal group, the the H2O2 model group, the apoptosis rate was significantly higher (P lt; 0.01); relative model group, lentinan concentration protection group cell apoptosis rate, there is a statistically significant difference significance (P lt; 0.01). Lentinan Caspase-3 protein expression H2O2 injury in SH-SY5Y cells cells after treatment, detection of Caspase-3 protein expression by Western blot method. Experimental results show that: the H2O2 model group cells Caspase-3 protein expression was significantly increased (P lt; 0.01); via lentinan 0.1,1,10 mg / L after treatment, cells Caspase-3 protein expression decreased significantly (P lt; 0.01 ).
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