Effect and mechanism of fuzhisan on toxic reaction of amyloid beta 25-35 to neural cells

Shirong WEN; Desheng WANG; Jingyan ZHANG; Shuli SHENG

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.