Mechanism Study Of Compatibility Of Aconitine And Ginsenoside Rg1 On Cerebral Cortex Neuron Cells

Objective:To research the minimum toxic concentration and the toxic mechanism of aconitine on cortex neuron cells, and whether the Ginsenoside Rgl can antagonise toxicity of aconitine or not.Methods:Uniform design and MTT method were used to study how aconitine of different concentrations influence on neuron cells at different time, and to find out the minimum toxic concentration and the critical role of time; the MTT method was used to determine how the mixture of aconitine and Ginsenoside Rgl in different proportions infect the survival of nerve cell; optical microscopy and electron microscopy were used to observe the influence on cell morphology by aconitine Ginsenoside Rgl and the mixture; enzyme method, turbidimetric method and ELISA were used to determine the influence on cell membrane, internal enviroment, energy metabolism and some of neurotransmitters, and to reveal the toxic mechanism of aconitine on nerve cells, then to explore whether the Ginsenoside Rgl can antagonise toxicity of aconitine or not.Results:The minimum toxic concentration of aconitine on nerve cell was 0.2%, and the critical role of the times effect on nerve cells were 0.5min,5min and 30min. After 30min,0.2% aconitine lead to the obvious broken and reduction of neurites, swelling of the cell body, incomplete of the membrane, emerging a large number of cytoplasmic vacuoles, loss of organelles, nuclear condensation, serious accumulation and margination of nuclear chromatin. The activity of LDH in culture medium was increased. In the cells, the activity of ACP, Na+, K+-ATPase and the contents of MDA, [K+] and [Ca2+] were increased, the activity of SOD and Ca2+,Mg2+-ATPase and the contents of [Na+] and [Mg2+] were decreased. The content of intracellular glycogen and the activity of COX were reduced. Meanwhile, the release of Glu and Ach within 24h was decreased, and the release of y-GABA, CA, SP and OP was increased. The mixture of aconitine and Ginsenoside Rgl in different proportions could lighten the changes caused by aconitine.Conclusions:0.2% aconitine had significant toxicity. It directly damaged the cell morphology and cell organelles, and also could cause damage of cell membrane, disturbance of internal enviroment, obstacle of energy metabolism and abnormal secretion of neurotransmitters. The toxicity mechanism of aconitine on nerve cells probably conducted in following way:aconitine damaged the biomembrane, and compensatory increased the activity of Na+, K+-ATPase. Ion concentrations between the inside and outside of cells were changed, which lead to obstacle of energy metabolism, changes of bioelectric potential and electrical signal, and abnormal release of neurotransmitters. Meanwhile, aconitine reduced the activity of Ca2+,Mg2+-ATPase, which lead to overload of intracellular calcium. As a result, cell morphology and function were damaged, and release of neurotransmitters was changed. Interaction of a variety of pathological conditions has led to aggravated injury, and ultimately manifested in the body for a variety of symptoms of poisoning.Ginsenoside Rgl could antagonise the toxicity of aconitine, the antagonism was probably to protect the cell membrane, inhibit the overload of intracellular calcium, improve the energy metabolization and regulated the release of neurotransmitters. Besides, the mixture of the two in equal proportion can gain the best effect.