Aim: This study aimed to investigate the protective effects of flavonoids from the stem and leaves of Scutellaria baicalensis Georgi (SSFs) against Aβ<sub>1-42</sub>-induced oligodendrocytes (OL) damage. ...Aim: This study aimed to investigate the protective effects of flavonoids from the stem and leaves of Scutellaria baicalensis Georgi (SSFs) against Aβ<sub>1-42</sub>-induced oligodendrocytes (OL) damage. Methods: Immunofluorescence was used for the detection of myelin-associated glycoprotein (MAG), a characteristic protein of rat oligodendrocytes (OLN-93 cells). To evaluate the potential protective effects of SSFs on OLN-93 cells injured by Aβ<sub>1-42</sub>, an injury model was established by subjecting OLN-93 cells to Aβ<sub>1-42</sub> exposed. Cell morphology was examined using an inverted microscope, while cell viability was assessed using the colorimetric method of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Additionally, lactate dehydrogenase (LDH) was measured using the pyruvic acid reduction assay. The Ginkgo biloba leaf extract (GBE) injection was used as a positive control. Results: A total of >95% of the MAG immunofluorescence-positive cells were identified as oligodendrocytes. Gradually increasing concentrations of SSFs impaired the cells, and the maximum nondetrimental dose for OLN-93 cells was 75 mg/L. This study assessed the effects of SSFs on OLN-93 cells damaged by Aβ<sub>1-42</sub>. The results indicated that SSFs significantly improved OLN-93 cell morphological abnormal changes, increased the OLN-93 cell survival rate, and reduced LDH release. Conclusion: SSFs can alleviate Aβ<sub>1-42</sub>-induced damage of OL.展开更多
BACKGROUND: L-3-n-butylphthalide (L-NBP) can inhibit phosphorylation of tau protein and reduce the neurotoxicity of beta-amyloid peptide 1-42 (Aβ1-42). OBJECTIVE: To observe the neuroprotective effects of L-NBP...BACKGROUND: L-3-n-butylphthalide (L-NBP) can inhibit phosphorylation of tau protein and reduce the neurotoxicity of beta-amyloid peptide 1-42 (Aβ1-42). OBJECTIVE: To observe the neuroprotective effects of L-NBP on caspase-3 and nuclear factor kappa-B (NF- K B) expression in a rat model of Alzheimer's disease. DESIGN, TIME AND SETTING: A cell experiment was performed at the Central Laboratory of Provincial Hospital affiliated to Shandong University between January 2008 and August 2008. MATERIALS: L-NBP (purity 〉 98%) was provided by Shijiazhuang Pharma Group NBP Pharmaceutical Company Limited. Aβ1-42, 3-[4,5-dimethylthiazolo-2]-2,5 iphenyltetrazolium bromide (MTT), and rabbit anti-Caspase-3 polyclonal antibody were provided by Cell Signaling, USA; goat anti-choactase and rabbit anti-NF- kB antibodies were provided by Santa Cruz, USA. METHODS: Primary cultures were generated from rat basal forebrain and hippocampal neurons at 17 or 19 days of gestation. The cells were assigned into five groups: the control group, the Aβ1-42 group (2 μmol/L), the Aβ1-42 + 0.1 μmol/L L-NBP group, the Aβ1-42 + 1 μ mol/L L-NBP group, and the Aβ1-42 + 10μmol/L L-NBP group. The neurons were treated with Aβ1-42 (2 μmol/L) alone or in combination with L-NBP (0.1, 1, 10 μmol/L) for 48 hours. Cells in the control group were incubated in PBS. MAIN OUTCOME MEASURES: Morphologic changes were evaluated using inverted microscopy, viability using the M-I-I- method, and the changes in caspase-3 and NF- k B expression using Western blot. RESULTS: Induction with Aβ1-42 for 48 hours caused cell death and soma atrophy, and increased caspase-3 and NF- K B expression (P 〈 0.05). L-NBP blocked these changes in cell morphology, decreased caspase-3 and NF- k B expression (P 〈 0.05), and improved cell viability, especially at the high dose (P 〈 0.05). CONCLUSION: AI3^-42 is toxic to basal forebrain and hippocampal primary neurons; L-NBP protects against this toxicity and inhibits the induction of caspase-3 and NF- K B expression.展开更多
文摘Aim: This study aimed to investigate the protective effects of flavonoids from the stem and leaves of Scutellaria baicalensis Georgi (SSFs) against Aβ<sub>1-42</sub>-induced oligodendrocytes (OL) damage. Methods: Immunofluorescence was used for the detection of myelin-associated glycoprotein (MAG), a characteristic protein of rat oligodendrocytes (OLN-93 cells). To evaluate the potential protective effects of SSFs on OLN-93 cells injured by Aβ<sub>1-42</sub>, an injury model was established by subjecting OLN-93 cells to Aβ<sub>1-42</sub> exposed. Cell morphology was examined using an inverted microscope, while cell viability was assessed using the colorimetric method of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Additionally, lactate dehydrogenase (LDH) was measured using the pyruvic acid reduction assay. The Ginkgo biloba leaf extract (GBE) injection was used as a positive control. Results: A total of >95% of the MAG immunofluorescence-positive cells were identified as oligodendrocytes. Gradually increasing concentrations of SSFs impaired the cells, and the maximum nondetrimental dose for OLN-93 cells was 75 mg/L. This study assessed the effects of SSFs on OLN-93 cells damaged by Aβ<sub>1-42</sub>. The results indicated that SSFs significantly improved OLN-93 cell morphological abnormal changes, increased the OLN-93 cell survival rate, and reduced LDH release. Conclusion: SSFs can alleviate Aβ<sub>1-42</sub>-induced damage of OL.
基金Supported by:the Medicine and Health Scientific Research Projects of Shandong Province,No. 2007HZ065
文摘BACKGROUND: L-3-n-butylphthalide (L-NBP) can inhibit phosphorylation of tau protein and reduce the neurotoxicity of beta-amyloid peptide 1-42 (Aβ1-42). OBJECTIVE: To observe the neuroprotective effects of L-NBP on caspase-3 and nuclear factor kappa-B (NF- K B) expression in a rat model of Alzheimer's disease. DESIGN, TIME AND SETTING: A cell experiment was performed at the Central Laboratory of Provincial Hospital affiliated to Shandong University between January 2008 and August 2008. MATERIALS: L-NBP (purity 〉 98%) was provided by Shijiazhuang Pharma Group NBP Pharmaceutical Company Limited. Aβ1-42, 3-[4,5-dimethylthiazolo-2]-2,5 iphenyltetrazolium bromide (MTT), and rabbit anti-Caspase-3 polyclonal antibody were provided by Cell Signaling, USA; goat anti-choactase and rabbit anti-NF- kB antibodies were provided by Santa Cruz, USA. METHODS: Primary cultures were generated from rat basal forebrain and hippocampal neurons at 17 or 19 days of gestation. The cells were assigned into five groups: the control group, the Aβ1-42 group (2 μmol/L), the Aβ1-42 + 0.1 μmol/L L-NBP group, the Aβ1-42 + 1 μ mol/L L-NBP group, and the Aβ1-42 + 10μmol/L L-NBP group. The neurons were treated with Aβ1-42 (2 μmol/L) alone or in combination with L-NBP (0.1, 1, 10 μmol/L) for 48 hours. Cells in the control group were incubated in PBS. MAIN OUTCOME MEASURES: Morphologic changes were evaluated using inverted microscopy, viability using the M-I-I- method, and the changes in caspase-3 and NF- k B expression using Western blot. RESULTS: Induction with Aβ1-42 for 48 hours caused cell death and soma atrophy, and increased caspase-3 and NF- K B expression (P 〈 0.05). L-NBP blocked these changes in cell morphology, decreased caspase-3 and NF- k B expression (P 〈 0.05), and improved cell viability, especially at the high dose (P 〈 0.05). CONCLUSION: AI3^-42 is toxic to basal forebrain and hippocampal primary neurons; L-NBP protects against this toxicity and inhibits the induction of caspase-3 and NF- K B expression.