Alcohol-induced Injury Of Spinal Cord Neurons In C57 Pups And Its Mechanism

Objectives: To investigate whether alcohol exposure causes damage to neurons in the ventral horn and dorsal horn of spinal cord in C57 pups and adults,we conducted a preliminary study.Methods: The 4th day mice pups after birth were randomly divided into two groups,named the control group and the alcohol group.The alcohol group was injected subcutaneously according to the body weight ratio,and the control group was injected with physiological saline according to the body weight ratio.Repeated 2 hours later;the same time as the 6th day of birth was injected again according to the above steps,and after 2 hours,the intact spinal cord tissue was dissected and retained.Spinal tissue was tested by immunohistochemical and immunofluorescence staining,Western Blot experiements.The other mice were tested in adulthood.Result: 1.Alcohol exposure can cause a significant increase in spinal cord Caspase 3positive cells(P<0.01),and spinal cord neuron apoptosis is enhanced.Immunohistochemical images showed that the expression of Caspase 3 was up-regulated in the ventral and dorsal horn of the spinal cord,and immunofluorescence images showed that alcohol caused an increase in the number of neuronal apoptosis in PD6 mice.2.Alcohol exposure can lead to a decrease in the number of Ch AT-positive neurons in the ventral horn of the spinal cord and a decrease in the number of GAD67-positive neurons in the dorsal horn of the spinal cord.The results of immunofluorescence double labeling showed that the damaged cell types were Ch AT-positive and GAD67-positive neurons.Immunohistochemical images also showed the distribution of Ch AT and GAD67.Western Blot showed the gray value of Ch AT and GAD67 decreased(P<0.05).3.Alcohol exposure can cause the expression of GFAP and Iba1 in the spinal cord of the rats to rise,that is,alcohol causes the inflammatory reaction of the spinal cord of the offspring,and the glial cells are activated.In immunohistochemical images,the denser GFAP-positive neurons were diffusely distributed in the spinal cord of the alcohol group(P<0.05),while the immunofluorescence images were more obvious(P<0.01).The expression of Iba1 showed a more significant increase in immunohistochemistry and immunofluorescence images(P<0.01).4.Alcohol exposure can lead to a decrease in the number of spinal cord neurons in adult mice,and the immunohistochemical images have a clear reflection(P<0.01).5.Alcohol exposure can reduce the area of ChAT-positive neurons in the ventral horn of spinal cord in adult rats(P<0.01)and the number of GAD67-positive neurons in the dorsal horn of spinal cord(P<0.01).Western Blot also has the same effect.6.Alcohol exposure can lead to decreased expression of SUFU,Shh and Islet1 proteins in the spinal cord of pups and adult mice(P<0.05).Conclusions: 1.Alcohol causes apoptosis of spinal cord neurons in C57 pups,which is manifested by the decrease of cholinergic neurons and GABAergic neurons.2.Alcohol causes inflammation of the spinal cord of the pups,and glial cells proliferate.3.Alcohol causes a decrease in the number of spinal cord neurons in adult mice,which is a decrease in cholinergic neurons and GABAergic neurons.4.Alcohol-induced developmental disorders and decreased numbers of cholinergic neurons and GABAergic neurons may be related to the inhibition of the Sonic hedgehog signal pathway and down-regulation of Islet1.