Effects Of XFM On Calcium Content And Calcium Channel Of Synaptosomes In Rat Cerebral Cortex

The miniature pigs’ combined anaesthetic (XFM) is is featured with safely, excellent effectand less side effect. However, the anesthesia mechanism is not fully understood. This study was toexplain the mechanisms of the central nervous system from the perspective of calcium ion assecond messengers in synaptosomes. In order to study the effects of XFM on calcium kinetics andcalcium channel of synaptosomes in rat cerebral cortex, and to analyze the relationship betweencalcium ions and XFM, and for further understanding to reveal the interaction mechanism betweenwith synptosomes, calcium channel blockers were taken to block the N-, P-, and L-type calciumchannels, and the concentation of calcium ions was detected through calcium fluorescent probe.SD rats were needed. The discontinuous sucrose density gradient procedure was used toisolate synaptosomes from cerebral cortex to allow calcium kinetic study to be performed.Synaptosomal suspension were obtained and calcium Fluo-3/AM loaded before going on torespectively added Xylazine, Tiletamine and XFM combination. Then, the concentation of calciumions was detected through calcium fluorescent probe to study the effects of XFM on calciumkinetics of synaptosomes in rat cerebral cortex. Again, synaptosomal suspension were obtained,and in order to study the the effects of XFM on calcium channel of synaptosomes in rat cerebralcortex, Nifedipine, ω-eonotoxin GVIA, ω-agatoxin IVA were added respectively to block L-, N-,and P/Q-type calcium ion channels before going on to detect the intracellular calcium ionconcentration after drugs effectiveness. The results of this experiment are as follows:1. The effects of XFM and its main reagents on calcium kinetics of synaptosomes in ratcerebral cortex:Xylazine could facilitate calcium ion mobilization induced by KCl in isolated synaptosomesin rat cerebral, and could make a surge of calcium ion concentration in the synaptosomes. Thedifference of60μM group and15μM group was extremely significant compared with controlgroup (P﹤0.01). The difference of30μM group was not significant compared with control group(P﹥0.05). Tiletamine could facilitate calcium ion mobilization induced by KCl in isolatedsynaptosomes in rat cerebral, and could make a surge of calcium ion concentration in thesynaptosomes. The difference was extremely significant compared with control group (P﹤0.01).There was a positive correlation between calcium mobilization and the concentration of Tiletamine.XFM could facilitate calcium ion mobilization induced by KCl in isolated synaptosomes in ratcerebral, and could make a surge of calcium ion concentration in the synaptosomes. The difference was significant compared with control group (P﹤0.05). There was a positive correlationbetween calcium mobilization and the concentration of XFM.2. The effects of XFM and its main reagents on calcium channel of synaptosomes in ratcerebral cortex:When the L-, and P/Q-type calcium ion channel were blocked, the decrease of the group ofXylazine was extremely significant compared with control group (P﹤0.01). While there was nosignificant difference compared with control group (P﹥0.05) to the N-type calcium ion channel.When the L-, and P/Q-type calcium ion channel were blocked, the decrease of the group ofTiletamine was extremely significant compared with control group (P﹤0.01). While there was nosignificant difference compared with control group (P﹥0.05) to the N-type calcium ion channel.When the L-type calcium ion channel were blocked, the decrease of the group of XFMcombination was extremely significant compared with control group (P﹤0.01). When the L-typecalcium ion channel were blocked, the increase of the group of XFM combination was extremelysignificant compared with control group (P﹤0.01).Conclusion:1. Xylazine and Tiletamine could facilitate L-, and P/Q-type calcium ion channel, andpromote calcium ion mobilization in synaptosomes in rat cerebral, and could make a surge ofcalcium ion concentration in the synaptosomes.2. XFM combination mainly effected L-type calcium ion channel, and promote calcium ionmobilization in synaptosomes in rat cerebral, and could make a surge of calcium ion concentrationin the synaptosomes. P/Q-type calcium ion channel was an secondary effect, N-type calcium ionchannel was no positive effect.3. XFM combination could respectively facilitate L-, and P/Q-type calcium ion channel, andpromote calcium ion mobilization in synaptosomes in rat cerebral, and could make a surge ofcalcium ion concentration in the synaptosomes. The release of excitatory neurotransmitter from thesynaptic vesicles increased, and caused anesthetic effect.