| Objective: We investigate the effect of monalsialic acid four hexose ganglioside sodium(GM-1)on bupivacaine-induced spinal neurotoxicity in rats on the basis of the establishment of bupivacaine-induced spinal neurotoxicity model,to explore the impact of bupivacaine-induced spinal neurotoxicity on spinal cord toxicity on endoplasmic reticulum stress related apoptosis factor of caspase-12,and to reveal the role of the pathway of endoplasmic reticulum stress apoptosis in the spinal neurotoxicity induced by bupivacaine,so as to provide basic data for the prevention and treatment of spinal neurotoxicity induced by the administration of local anesthetics in clinical practice.Methods: A total of 108 healthy adult male SD rats were selected,and the modified Yaksh method was used in sheath built-in tube to the tail end of the L3-4.The rats with successful puncture were divided into the control group,the Bupivacaine group and the GM-1 group according to the random number method.In the Bupivacaine group,0.12?l/g 5% bupivacaine was injected intrathecally for a total of three times with the interval of 1.5h,and 6 rats each were taken after the administration on 0d,1d,3d,5d,7d,14 d,respectively.Ratsin GM-1 group received intrathecal pre-injection of 20?g GM-1 before intrathecal injection of 5% bupivacaine,and 6 rats each were sampled after 5%bupivacaine injection on 0d,1d,3d,5d,7d,14 d,respectively.Rats in the control group were provided with intrathecal injection of the same amount of normal saline only,without pre-injection GM-1.No bupivacaine toxicity model was established at the same time in the control group,and 6 rats each were taken after the administration on 0d,1d,3d,5d,7d,14 d,respectively.Determination of the tail flick latency(TFL)and calculation of the maximum permissible exposure(MPE)percentage were achieved before intrathecal injection and after the administration on 0d,1d,3d,5d,7d,14 d,followed by hindlimb motor function assessment by using the Basso Beattie Bresnahan(BBB)rating scale.Spinal cord tissue was stained with HE after TFL and BBB score,and histopathological changes were observed under light microscope.Furthermore,immunohistochemistry and qRT-PCR were used to determine the protein and mRNA expression of caspase-12 in the spinal cord tissue of each time point.Results: There was no neurological and histological changes at each time point of 0d,1d,3d,5d,7d,14 d in the control group.In comparison with the control group,the TFL was prolonged,the percentage of MPE was increased significantly,and the BBB score was decreased at the same time point of 0d,1d,3d,5d,7d,14 d in the Bupivacaine group.Furthermore,mRNA and protein expression levels of caspase-12 were increased in varying degrees(P<0.05),associated with obvious pathological injury of spinal cord tissues.Meanwhile,compared with the control group at the same time points of 0d,1d,3d,5d,7d,14 d,the percentage of MPE was increased,the BBB score was decreased,and caspase-12 mRNA and protein expression were increased synchronously inGM-1 group(P<0.05).In addition,the percentage of MPE was clearly decreased,and the BBB score was increased evidently when compared to the Bupivacaine group at the same time points of 5d,7d,14d(P<0.05);besides,the mRNA and protein expression of caspase-12 were decreased synchronously(P<0.05),accompanied by reduced pathological injury in spinal cord tissues.Conclusion: The present study suggests that GM-l inhibits caspase-12 synthesis,decreases caspase-12 expression in injured spinal cord,and reduces or inhibits neuronal cells apoptosis,which is correlated with the inhibition of ERS induced neuronal cells apoptosis. |
PDF Full Text Request