Effect Of Hyperbaric Oxygen Auxiliary Antivenom On The Agkistrodon Acutus Envenomation Rat's SVZ

Objective: Through established the Agkistrodon acutus venom rat's model,the present study observed the effect of Agkistrodon acutus venom on the Nestin gene and protein's expression in rat's SVZ and explored the effect of intervention with HBO combined antivenom on its expression.Methods: 96 male Sprague-Dawley rats were randomly divided into 4 groups including control group,snake venom group,antivenom group,HBO+antivenom group.And there were 24 rats in each group.The poisoning model was established by injecting venom via a rat's caudal vein in snake venom group.Similarly,rats in antivenom group were immediately injected specific antivenom after successful envenomation model.Rats in HBO+antivenom group were exposed to HBO only once at 0 h,4 h,11 h and 23 h after antivenin injection,and 6 rats were grouped in each time point.Rats in control group were no treated.The brain tissue of part rats in the whole group was fixed through perfusing physiology saline and paraformaldehyde by left ventricle at 1 h,5 h,12 h,24 h after antivenom injection.And other parts were immediately separated SVZ in ice from the whole brain tissue and quick-frozen standby.HE was used to investigate the pathology changes of rat's brain in the whole group.IHC,RT-PCR,and Western blot were used to detected the expression of Nestin m RNA and protein in SVZ.Results: 1.Brain pathology changes: edema brain tissue,decreased cell number,and disordered arrangement cells that occurred in SVZ in snake venom group.The antivenom and HBO intervention significantly alleviated the edema tissue and increased the cell number with the regular arrangement,which was more prominent in HBO+antivenom group,especially within 5 h after envenomation.2.The expression of Nestin gene and protein: Nestin protein expresses on neural stem cell cytoplasm.The IHC showed that no Nestin positive cells were expressed in control group,and only a few positive ones in snake venom group.The positive cells were significantly increased and color deepened in antivenom and HBO+antivenom groups(P<0.05).The relative quantitative results show that compared with control group,Nestin m RNA and protein's expression of snake venom group at each time point in SVZ was significantly elevated(P<0.05).Compared with venom group,Nestin gene and protein's expression was more evident in antivenom group and HBO+antivenom group at 1 h and 5 h after venom injection(P<0.05).And expression in HBO+antivenom group was more obvious than that in antivenom group(P<0.05).Conclusion: 1.Agkistrodon acutus venom has neurotoxic effect on brain.2.The administration of HBO combined with antivenom show neuroprotection after Agkistrodon acutus envenomation,which may be related to the increased expression of Nestin protein in SVZ.3.The neuroprotection of HBO early intervention combined antivenom is superior to antivenom intervention,and the role is most significant especially within 5 h after venom injection.