| Objective The experiment was aimed at investigating the role which lipid emulsion plays towards NMDAR and GABA_AR as in the neurotoxicity model stimulated by bupivacaine to confirm whether lipid emulsion could alleviate bupivacaine-induced central nervous toxicity through the regulation of the expression of NMDAR and GABA_AR.Methods 1.20 healthy male SD rats,280 g-320 g,were randomly divided into two groups: bupivacaine+normal saline group(BPV + NS group),bupivacaine+lipid emulsion group(BPV+LE group).Animals were scheduled to seizure by injecting bupivacaine through caudal veins to make the convulsion model.NS or LE were infused right after the aforementioned model.Keep an observation of their Tarlov scores within 1 hour after treated with NS or LE.2.The above-mentioned healthy SD rats,amount to 54,were divided into three groups at random: control group(sham),BPV + NS group,BPV+LE group.The latter two groups were conducted as has been noted,while the control group were pumped with the same volumes of normal saline.It was until 10min(or 30min?1h?2h?6h?12h)after injected with NS or LE did 3 randomly-selected rats in each group be sacrificed for detecting Glu with HPLC.3.After careful analysis about the content of Glu,it can be inferred that rats which were killed 6h after NS or LE seem to be statistically valuable.18 healthy SD rats were assigned to three groups the same as previous.Six hours after being treated with NS or LE,rats' s head were cut off to take out the brain tissue and the levels of CaM as well as NMDAR and GABA_A were tested by Immunohistochemical method or Western Blot respectively.4.21 healthy SD rats,as stated above,were allocated to seven groups: sham,BPV + NS group,BPV+LE group,bupivacaine+ normal saline+ MK-801 group(BPV +NS+MK-801),bupivacaine+lipid emulsion+MK-801 group(BPV +LE +MK-801),bupivacaine+normal saline+L-NAME group(BPV+NS+L-NAME),bupivacaine+lipid emulsion+L-NAME group(BPV + LE+L-NAME).The prior period is of quite the same.The added four groups were handled with MK-801 or L-NAME through intraperitoneal injection one hour after NS or LE infusion.The activity of nNOS in rats' hippocampus were measured after the time point as previously mentioned.Results 1.Tarlov scores of group BPV+LE were higher than those of group BPV+NS after treated for 3min,5min,10min(P<0.01).2.The content of Glu in group BPV+NS outnumber group BPV+LE under the circumstance that the seizure model were made 1h,2h,6h prior to death(P<0.05),among which 6h-group holds the maximum.3.Throughout the animals which were killed at the time-point of 6 hours,the positive area of CaM in group BPV+LE yielded to group BPV+NS(P<0.05).Levels of NMDAR in group BPV+LE were inferior to group BPV+NS,the difference was statistically significant(P<0.01).The expression of GABA_AR in group BPV+LE was higher than that of the BPV+NS group(P<0.05).4.The activity of nNOS in the BPV+NS/LE+MK-801 group exceeded the sham group and its corresponding model group(P<0.05).No statistical difference occured in other groups.Conclusion Lipid emulsion can alleviate the neurotoxicity induced by bupivacaine through the regulation of NMDA-NO pathway and GABA_AR. |
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