| Objective: It is a pressing social and medical problem to find effective drugs for deferring aging process and good approaches for prevention and cure of brain diseases. In the view of Traditional Chinese Medicine, brain is the sea of marrow, and the kidney stores essence of life and promotes the formation of marrow. The essence stored in kidney is the important foundation for the function of brain. The spleen provides the material basis for the acquired constitution, and is necessary for the function of kidney. For the insufficiency of spleen and stomach of senior people, it is difficult to prevent brain aging by the way of replenishing essence of kidney directly. So replenishing essence of kidney and reinforcing spleen at the same time is an effective and reasonable approach to deferring aging. The function of spleen is implemented by the cooperation of spleen-Yin and spleen-Yang. Spleen-Yin includes the Yin fluid( blood, body fluid, refined materials of food and water) , material morphology and nourishing function of spleen。Insufficiency of spleen-Yin is an important part of spleen deficiency syndrome. Food and water will not be digested with deficient spleen-Yin and the insufficiency of Yin may cause unnourished blood and vessels, Result in the malnutrition of congenital essence. Consequently, the deficiency or disease may lead to aging. So, to reinforce the Yin of spleen is a good approach to defer aging. In central nervous system,Glutamate (Glu) is the most abundant excitatory amino acid and the most important excitatory neurotransmitter which is the basic of learning and memory. Neurons will be damaged when Glu receptors are over-activated. Extracellular Glu abnormal accumulation would lead to overstimulation of NMDAR, and this overstimulation brings neurons to the excitotoxic injury or cell death. Therefore, anti-excitotoxicity become a very important strategy for neuroprotection and postponing brain aging. Our past study showed that Zi-Bu-Pi-Yin (ZBPY) recipe could enhance the ability of learning and memory in different aging animal models, protect mitochondria membrane from lipidsupereoxyde, increase activity of mitochondria respiration enzyme-cytochrome oxidase enzyme, remain the rate of phospholipid, improve energy dymatabolism and have the protective effect on neurons damaged by excitatory neurotransmitter. So it has apparent role of deferring aging. Our study aims at evaluating protective effects of ZBPY recipe on hippocampal neurons in excitotoxicity system model, furthermore exploring the deferring decrepitude mechanisms of ZBPY recipe. Method:We established Glu excitotoxicity system model of primary hippocampal neurons with 100μM Glu and 10μM glycine on the 12th day. Experiments presented here included ZBPY recipe pretreatment group, cholesterol pretreatment group, ZBPY+chlolesterol pretreatment group, simvastatin pretreatment group and MK-801 positive control group which generally accepted as anti-excitotoxicity drug. The degree of neuron damage was evaluated by trypan blue drying and detection of LDH efflux in supernatant. The protective effect of ZBPY recipe drug serum (5%) on neurons damaged by Glu was studied by serum pharmacology. Intracellular cholesterol was detected by High Performance Liquid Chromatography (HPLC) and the cholesterol in supernatant was detected with enzymatic method, the results were used to analysis the positive effects onhomeostasis of cholesterol in excitototic injury neurons, and simvastatin pretreatment group was took as positive control group which decreased cholesterol in neurons in advance . Result: 1. Cell viability: Cell viability of Glu group was 68.75±3.43% , the cell viability of ZBPY recipe group was 86.35±1.43%.Comparing with the Glu group, cell viability of ZBPY recipe pretreatment group increased significantly(p<0.01).the cell viability of MK-801 group was 94.42±1.50%. The cholesterol group neurons'cell viability was 57.42±3.13%. The cell viability of ZBPY+cholsterol group was 72.23 ±1.53%. Comparing with Glu group, differences between them...
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