Effects of drug serum in broken bushen yizhi formulas on cell model of Alzheimer disease

Yunbo CHEN; Shilong LAI; Jingqing HU; Qi WANG; Shuyi CHENG

ABSTRACT

BACKGROUND:

Chinese herb, bushen yizhi formula protects at certain extent learning and memory in rat model of Alzheimer disease. The drug serum in this formula can alleviate neurotoxic reaction of nerve tumor cell NG 108-15 to beta-amyloid protein. In order to understand further the mechanism and compatibility of the formula, it is necessary to carry on the study on the broken formulas.

OBJECTIVE:

To study the effect of drug serum in subgroups of broken bushen yizhi formulas on growth and differentiation of cell model of Alzheimer disease and probe into the compatibility rule of bushen yizhi formula in view of serum pharmacology.

DESIGN:

Randomized controlled experiment.

SETTING:

DME Center of Clinical Pharmacological Institute Affiliated to Guangzhou University of Traditional Chinese Medicine.

PARTICIPANTS:

The experiment was performed in DME Center of Clinical Pharmacological Institute Affiliated to Guangzhou University of Traditional Chinese Medicine from January to August 2003, in which, 40 healthy male SD rats of 3 months old were employed and NG108-15 cell line was frozen-preserved.into the control, original formula group (No. 1 group) [shechuanzi (Cnidium monnieri (L.) Cuss), gouqizi (Lycium barbarum L.), renshen (Panax ginseng C.A.Mey.), heshouwu (Polygonum multiflorum Thunb.), danpi (Paeonia Suffruticisa Andr.) and bingpian (Borneolum)], kidney replenishment group (No.2 group) [shechuanzi (Cnidium monnieri (L.) Cuss), gouqizi (Lycium barbarum L.), etc.], group for benefiting qi and nourishing blood (No.3 group)[renshen (Panaz ginseng C.A.Mey), zhishouwu (Polygonum multiflorum Thunb.), etc.] and group with bingpian (Borneolum) removed (No.4 group)[Panax ginseng C.A.Mey.], heshouwu (Polygonum multiflorum Thunb.) and danpi (Paeonia Suffruticisa Andr.)], 8 rats in each group. The concentrated Chinese herbal solutions of every group were applied at 10 μL/g (equal to 6 g/kg of raw herbs) for gastric infusion successively,continuously for 1 month. In the control, the physical saline solution of equal dosage was used for infusion. Two hours after the last gastric infusion in rats of each group,the blood was collected from heart after anesthesia and the serum was sepaNG108-15 cell cultured in vitro was divided into 6 groups. In the control and model group, normal rat serum was contained in proliferated culture solution. In the rest 4 groups, the drug serum of No. 1 group and 3 sub-groups was contained.Simultaneously, beta-amyloid protein 25-35 in each hole was prepared to the terminal concentration 5 μmol/L (except in the control) and the culture went on for 48 hours.MAIN OUTCOME

MEASURES:

MTT method was used to determine proliferated number and survival rate of cells. Simultaneously, the ratio of neurite cells to total cell count and average length of neurit were determined.icantly than the control (0.520±0.022, 0.665±0.037, P ＜ 0.01), and that in every drug serum group was higher than model group, of which, the result vival rate of differentiated cells That in model group was lower significantly than the control (58.4%, 100%) and that in every drug serum group was higher than model group, of which, the result in No.4 group was the most tal cell count That in model group was lower significantly than the control [(42.95±11.42)%, (58.75±12.84)%, P ＜ 0.01] and that in every drug serum group was higher than model group, of which, the result in No.4 group was rite That in model group was shorter significantly than the control [(356.0 ±109.0), (493.8±133.0) μm, P ＜ 0.01] and that in every drug serum group was longer than model group, of which, the result in No.4 group was the most significant [(486.8±79.2) μm, P ＜ 0.01].

CONCLUSION:

The drug serum in all of bushen yizhi formula and every subgroup inhibits at certain extent the injury of beta-amyloid protein 25-35 to NG108-15 cell, but the results of each group are various. The protection of drug serum to the cell in every group is in the sequence from strong to weak as group with bingpian removed ＞ original formula group ＞ kidney replenishment group ＞ group for benefiting qi and nourishing blood. It is to expect a further study on the efficacy of group with bingpian removed.