Joint Study On Voltage Threshold Method And High Content Technology For Influence Of Morphine Hydrochloride And Lidocaine Hydrochloride On Neural Networks

The functions of various organs and systems of the human body are directly or indirectly regulated and controlled by the nervous system.As the whole body of the controller and the coordinator,the phenomenon which some medicine would produce side effects to nervous system in the clinical treatment is not uncommon,such as some drugs can aggravate the symptoms of the original nervous system diseases,other performance is similar to the nervous system diseases or symptoms of mental illness.In order to safe,reasonable and effective use of drugs,it is important to establish an efficient drug screening platform for quantitative analysis of the effects of various drugs on neural networks.The voltage threshold mesuating method proposed by our group can quantitatively evaluate the influence of the external factors to excitability of the neural network,and we already have used this method to study the effects of alcohol,acetylcholine and temperature on the neural network.In this paper,we proceed from the point of drugs and narcotics and selected morphine hydrochloride and lidocaine hydrochloride as the target drugs,respectively.The nerve tissue(rat hippocampal slices)and primary cultured neural network(rat hippocampal neuron networks)as networks model.Our aims are to quantitatively analyze the effects of two drugs on the excitability of hippocampal neurons and hippocampal neurons,and obtaine the voltage threshold of neural networks.Then,high-content cellomics analysis system was used to study the effect of two drugs on the apoptosis and the length of the neuron networks.The results were compared and analyzed with the results of voltage threshold in conjunction to reveale the mechanism of the effect of two drugs on the neural network voltage threshold.In this study,rat hippocampal slices and rat hippocampal neuron networks were cultured on MEA.Under the action of morphine hydrochloride and lidocaine hydrochloride,we conducted the response signal detection experiment to neural networks and obtained the voltage threshold of the networks.Then,we quantitatively analyzed the electrical excitability of two kinds of neuron networks by different concentrations of morphine hydrochloride and lidocaine hydrochloride according to the curve of voltage threshold-concentrations of drugs curve.The results showed that:1)Under the different concentrations of morphine hydrochloride,the voltage threshold of rat hippocampal slices and rat hippocampal neurons was higher than the standard voltage threshold,which indicated that morphine hydrochloride inhibited the electrical excitability of neural networks,and the concentration of morphine hydrochloride was positively correlated with the inhibitory effect to neural network of excitability.2)lidocaine hydrochloride had the effects of biphasic effects on the electrical excitability of rat hippocampal neurons.In the concentration range of 0.1-0.5 ?g/mL,there was a positive correlation between the concentration and the voltage threshold,and the inhibitory effect of lidocaine hydrochloride was the strongest when the concentration is 0.5?g/mL;In the concentration range of 0.5?1.1?g/mL,lidocaine hydrochloride still would be inhibitory effect,but the inhibition effect was weakened;In the concentration range of 1.1?1.3?g/mL,the effect of lidocaine hydrochloride on the excitability of rat hippocampal neuron network is excited,and with the increase of the concentration of lidocaine hydrochloride,the voltage threshold of the neuron networks continued to decrease.3)By contrasting the relationship between the threshold values of rat hippocampal slices and rat hippocampal neurons with the action of morphine hydrochloride and lidocaine hydrochloride,respectively,it is found that the voltage thresholds of the two neural networks were consistent and confirmed.4)By contrasting effects of morphine hydrochloride and lidocaine hydrochloride on the excitement of neural networks to the same neural network model(rat hippocampal neural network,it was found that the inhibitory effect of morphine hydrochloride on neural networks was stronger than that of lidocaine hydrochloride.Finally,the rat hippocampal neurons were irradiated with different concentrations of two drugs and used the high content screening technique to analyze the effects of drugs on neuronal apoptosis and protrusions.The results showed that:1)Morphine hydrochloride and lidocaine hydrochloride did not cause apoptosis in the concentration range of this paper,and had no toxic effect on the neural networks;2)Morphine hydrochloride changed the length and area of the hippocampal neuron networks,and this change was negatively correlated with the concentration of morphine hydrochloride.And this trend of the length of the protrusion was opposite to the trend of the voltage threshold.3)The concentration of lidocaine hydrochloride was 0?0.5?g/mL,the length and area of neurons was negatively correlated with the concentration of lidocaine hydrochloride;When the concentration increased from 0.5 to 1.4?g/mL,the length and area of the neuron networks showed increasing trend,which is contrary to the trend of voltage threshold variation.Because the sodium ion channel dense band on the nerve cell excitability has a regulatory role,the role of drugs led to changes in the electrical excitability of neurons.The position and length of the "sodium band" have a moderating effect on the excitability of the neurons.It is presumed that the effects of the two drugs on the neuron protrusions change,leading to changes in the "sodium band" located on the axons of the neurons,The electrical excitability of the element changes and is manifested by a change in the voltage threshold.