The Effect Of Huoxuetongdu Decoction On Motor Function Of Hind Limb And Apoptosis In Rabbits With Spinal Cord Ischemia/Reperfusion Injury

Objective Observing the effect of Huoxuetongdu decoction on motor function of hind limb and the expression of c-fos and c-jun, which is apoptosis related gene in rabbits with spinal cord ischemia/reperfusion injury and further explore the protection mechanism of the Huoxuetongdu decoction on spinal cord ischemia reperfusion injury.Methods1.Jacobs score of hind limb function of rabbits:New Zealand rabbits（n=24） weighting2.0±0.2kg of both genders were equally and randomly assigned to a sham surgery group（n=8）,spinal cord ischemia/reperfusion injury（SCⅡ） group（n=8） and Huoxuetongdu decoction（HD） group（n=8）. Rabbits in the sham surgery group and SCII group were intragastrically administered saline for7consecutive days before operation, and for21consecutive days after operation. In the HD group, the rabbits were intragastrically administered HD before operation for7consecutive days, and for21consecutive days after operation. Models of spinal cord ischemia/reperfusion injury were established in the SCII group and HD group by separating and occluding L3-6for30minutes, followed by reperfusion. In the sham surgery group, only L3₆was separated. Hind limb motor function was observed and scored using Jacobs method at8and24hours, and1,2and3weeks after reperfusion.2. Mechanism: New Zealand rabbits（n=96） weighting2.0±0.2kg of both genders were equally and randomly assigned to a sham surgery group（n=32）, spinal cord ischemia/reperfusion injury（SCII） group（n=32） and Huoxuetongdu decoction（HD） group（n=32）. In each group was divided into reperfusion0.5h,1h,4h,8h4subgroups, each subgroup had8rabbits. Rabbits in the sham surgery group and SCII group were intragastrically administered saline for7consecutive days before operation. In the HD group, the rabbits were intragastrically administered HD before operation for7consecutive days. Models of spinal cord ischemia/reperfusion injury were established in the SCII group and HD group by separating and occluding L3₆for30minutes, followed by reperfusion. In the sham surgery group, only L3.6was separated. Each group derived after reperfusion0.5h,1h,4h,8h, a total of four time points. The levels of c-fos and c-jun protein content in the spinal cord tissue was measured by Western-blot. The number of c-fos and c-jun positive cells in the spinal cord tissue was measured by immunohistochemistry.Results1. Hind limb motor function was normal in the sham surgery group at various time points after reperfusion, and no significant changes were detected between preoperation and postoperation. Jacobs score was lower in the SCII and HD groups than that in the sham surgery group （P<0.01）. At8hours after reperfusion, Jacobs score was reduced in the SCII and HD decoction groups, especially in the SCII group （P<0.01）. At8-24hours after reperfusion, the recovery of hind limb motor function was slow in the SCII and HD groups. At24hours after reperfusion, significant differences in the recovery of hind limb motor function were detectable between the SCII and HD decoction groups （P<0.01）. With prolonged reperfusion time, the recovery of hind limb motor function was not obvious in the SCII group, but noticeable in the HD group at24hours-3weeks. Significant differences in Jacobs score were detected between the SCII and HD groups at1,2and3weeks （P<0.01）.2. C-fos and c-jun protein lightly expressed in the sham surgery group at0.5,1,4, and8hours. C-fos and c-jun expression increased at0.5hour in the SCII and HD groups, and reached a peak at1hour, and then diminished. At the same time points, c-fos and c-jun expression was higher in the SCII and HD groups compared with sham surgery group （P<0.01）. C-fos and c-jun expression was lower in the HD group than that in the SCII group at four time points （P<0.05or P<0.01）.Conclusions1. Huoxuetongdu decoction improved spinal cord ischemia/reperfusion injury, contributed to the recovery of hind limb motor function, and ameliorated spinal ischemia/reperfusion injury.2. Huoxuetongdu decoction ameliorated spinal cord ischemia/reperfusion injury possibly through inhibiting c-fos and c-jun protein expression in the spinal cord, and reducing nerve cell apoptosis.