ABSTRACT
BACKGROUND: Amyloid β(Aβ) protein is the core of senile plaque.Being the toxic segment of Aβ, Aβ25-35 has been extensively applied in the experiments of recent years. The research in the past has verified that the self-prepared Chinese herb, fuzhisan can promote the survival of the cultured neural cells and probably acts on the treatment of Alzheimer disease (AD).OBJECTIVE: To study the resistance of fuzhisan to Aβ25-35 toxicity to cultured neural cells and the probable approaches.DESIGN: Repeated measurement based on the cells.SETTING: Department of neurology of a university hospital and key experimental room in brain aging in a university hospital.MATERIALS: The experiment was performed in Beijing Key Experimental Room in Brain Aging of Xuanwu Hospital from June 2002 to April 2003. Dopaminergic SH-SY5Y cell of neuroblastoma and Aβ25-35 were employed. Chinese herb, fuzhisan was decocted with mild fire and its upper clear solution was collected and prepared into storage solution at the concentration of 0. 5 g/mL. Antibody: Beijing Key Experimental Room in Brain Aging of Xuanwu Hospital prepared cAMP responsive element binding protein(CREB),Bcl-2 in B lymphatic leukaemia-2 genetic product and cytochrome C(CytC).METHODS: SH-SY5Y cell was incubated with Aβ25-35 of various doses alone or in combination with fuzhisan and was compared with blank control. MTF metabolic rate of cultured neural cells were determined under different incubation conditions. Western-blot method was used to measure the protein expression changes in incubation with fuzhisanalone, incubation with Aβ25-35 alone and the combination incubation, compared with the blank control.MAIN OUTCOME MEASURES: It was to study MTT metabolic rate in the comparison between each experimental group and the blank control and expressions of CREB, Bcl-2 and CytC relevant to survival/death of neural cells.RESULTS: Survival rate of SH-SY5Y cell was increased by 11.4% in incubation with fuzhisan alone. It was remarkably improved in incubation combining fu zhisan with Aβ25-35 as compared with Aβ25-35 alone. The expressions of CREB and Bcl-2 in Aβ25-35 group were decreased and were increased in fuzhisan group. CytC expression in cytoplasm was increased in Aβ25-35 group and was declined with fuzhisan incubation.CONCLUSION: Fuzhisan promotes the survival of cultured neural cells and its protection is still existed under Aβ25-35 injury. Fuzhisan brings such effects into play probably by the protein expressions relevant to survival/death of cells.
ABSTRACT
BACKGROUND: It has been verified in the experiments over the past that the self-prepared Chinese herb, fuzhisan can retard natural aging in rats, suggesting that such drug acts on anti-aging.OBJECTIVE: To observe the effect of optimum effective concentration of nerve cell cultured with fuzhisan on morphological alternation of neuroblastoma SH-SY5Y cell.DESIGN: Repeated measurement.MATERIALS: The experiment was performed in Beijing Key Laboratory Room of Cerebral Aging of Xuanwu Hospital Affiliated to Capital University of Medical Sciences from June 2002 to April 2003. Self-prepared Chinese herb, fuzhisan [composed of 6 herbs, such as shichangbu (Rhizoma Acori Graminei), yuanzhi (Radix Polygalae), etc.] was co-developed by Prof.Wang De-shen from Department of Neurology of First Clinical Medical College of Harbin Medical University and Prof. Xu Xiao-yun from Department of Neurology of Shanghai Oriental Hospital. In addition, amyloid βprotein 25-35 segment and SH-SY5Y neuroblastoma were provided.METHODS: Fuzhisan of various concentrations were used for incubation of neuroblastoma SH-SY5Y cell. Thiazolyl blue (MTT), colorimetric method was used to determine the cell survival rate. Dose-effect relationship curve was drawn up to search optimum drug concentration. The cells cultured with 6-pore plate were divided into normal control, amyloid β-protein 25-35 25 μmol/L group, amyloid β-protein 25-35 25 μmol/L + fuzhisan 45×10-3 g/L groups and fuzhisan 45×10-3 g/L group. They were incubated for 24 hours to observe cell morphological alternation and determine neurosome area and axon length.ery group.length of every group: Those in amyloid β-protein 25-35 25 μmol/L group were decreased remarkably than the normal control [(505.5 ±122.36),(599.8 ±141.25) μm2; (26.0±13.97), (36.5 ±15.58) μm, (t =3.903,3.447, P=0.000)]. Those in fuzhisan 45×10-3g/L group and amyloid β-protein 25-35 25 μmol/L + fuzhisan 45×10-3 g/L group were increased remarkably than amyloid β-protein 25-35 25 μmol/L group [(918.3±178.34),(896.6 ±257.14), (505.5 ±122.36) μm2; (96.8 ±43.31), (88.3 ±30.23),(26.0±13.97) μm, (t=10.922, 14.172, P=-0.000)].CONCLUSION: With injury of amyloid β-protein 25-35, fuzhisan still enhances the survival of cultured nerve cell, manifested as promoting the increase of neurosome area and axonal extension.
ABSTRACT
<p><b>OBJECTIVE</b>The -455 G/A (HaeIII) polymorphism of beta-fibrinogen gene influences levels of plasma fibrinogen. We further investigated whether it influences the risk of ischemic cerebrovascular disease.</p><p><b>METHODS</b>We accumulated 134 acute ischemic cerebrovascular disease (ICVD) cases and compared their -455 G/A status with a control group (n = 166). The beta-fibrinogen gene -455 G/A polymorphism was analyzed for all subjects by PCR-RFLP with the restrictive enzyme HaeIII.</p><p><b>RESULTS</b>Plasma fibrinogen was higher in AA homozygous participants (341 mg/dL) than in participants carrying the G allele: GA (290 mg/dL), GG (298 mg/dL) in the control group. Plasma fibrinogen was also higher in AA homozygous patients (353 mg/dL) than in cases carrying the G allele: GA (287 mg/dL), GG (302 mg/dL) in the ICVD group. However, there was no significant association between beta-fibrinogen gene -455 G/A polymorphism and ICVD group.</p><p><b>CONCLUSIONS</b>Although a small effect cannot be excluded, beta-fibrinogen gene -455 G/A polymorphism is an independent predictor of plasma fibrinogen, but not of ischemic cerebrovascular disease.</p>