| Purpose:In this experiment,warm acupuncture therapy was applied to the spinal cord injury model rats to observe the behavior of the spinal cord injury model rats and the mRNA and protein expressions of Shh and Gli-1 in different time periods at the spinal cord injury site.The relationship between the effect of warm acupuncture on spinal cord injury repair and Shh/Gli-1 signal pathway was explored to explore the mechanism of warm acupuncture with on spinal cord injury and provide experimental basis and related theoretical basis for clinical warm acupuncture therapy on spinal cord injury.Methods:This experiment adopts the design method of random grouping and parallel control to carry out research.54 spinal cord injury rats models of the 10 th thoracic vertebra(T10)were prepared by referring to the modified Allen’s heavy hitting method,Then,54 rats with successful modeling were randomly divided into four groups according to envelope method,namely acute acupuncture warm acupuncture group,convalescent warm acupuncture treatment group,continuous warm acupuncture treatment group and model control group.Among them,continuous treatment group and model control group have two subgroups respectively,namely continuous warm acupuncture treatment group for 7 days,continuous warm acupuncture treatment group for 14 days,model control group for 7 days,and another group of normal blank control group,totaling 7 groups,each group has 9 rats,totaling 63 rats.Each treatment group was treated with warm acupuncture according to the division of different time periods.At the same time,BBB scoring method was used to score the motor function of both lower limbs of rats 24 hours,7 days and 14 days after the model was established.On the 7th day after modeling,the spinal cord samples of the 7-day model group and the 7-day continuous treatment group were obtained,while the spinal cord samples of the remaining groups were obtained after the treatment was completed on the 14 th day and BBB score was completed.After successful sampling,the expression of Shh mRNA and Gli-1 mRNA in each group of spinal cord injury rats was detected by real-time fluorescence quantitative PCR,and the expression of Shh and Gli-1 protein in each group of spinal cord injury rats was detected by Western blot.All experimental data were statistically analyzed by SPSS22.0 statistical software.when P≥0.05,there was no statistical significance;P<0.05,there was statistical significance;P<0.01,there was extremely significant statistical significance.Results:1.Observation on behavioral indexes of rats in each groupThe results of BBB scores of each group at 3 time points after operation showed that the BBB score of rats in each group was 0 on the 1st day after operation,and the modeling was successful.On the 7th day,the BBB score of the rats that had been treated with warm acupuncture was significantly higher than that of the model control group(P<0.05).On the 14 th day,the BBB scores of rats in all treatment groups were still higher than those in the model control group(P<0.05).compared with the other two treatment groups,the BBB scores in the convalescent treatment group were lower than those in the other two treatment groups,and P<0.05.the BBB scores in the continuous treatment group were slightly higher than those in the acute treatment group,but P>0.05.2.Observation on mRNA expression of Shh and Gli-1 in rats of each groupThe mRNA expression results of Shh and Gli-1 in rats of each group showed that compared with the normal control group,the mRNA expression of Shh and gli-1 in model control group and warm acupuncture treatment group were increased,and P<0.05.The expressions of Shh and Gli-1 mRNA in the continuous treatment group were higher than those in the model control group at the 7th and 14 th days,and P<0.05.There was no significant difference in Shh and Gli-1 mRNA expression between the 7-day model group and the 14-day model group(P>0.05).Although Shh mRNA expression in convalescent treatment group increased slightly compared with model 14 day group,but P>0.05.Shh mRNA expression in acute treatment group increased significantly compared with model 14 day group,P<0.05.The expression of Gli-1 mRNA in each treatment group was higher than that in the 14-day model group,and P<0.05.The expression of Shh and Gli-1 mRNA in acute treatment group was higher than that in convalescent treatment group,but lower than that in continuous treatment group for 14 days,the difference was statistically significant(P<0.05).3.Observation on protein expression of Shh and Gli-1 in rats of each groupThe protein expression results of Shh and Gli-1 in different periods of time in each group showed that there was no significant difference in Shh and Gli-1 protein expression between the 7-day model group rats and the normal control group rats(P>0.05);The expression of Shh and Gli-1 protein in the 14-day model group was significantly higher than that in the normal group(P<0.05).The expression of Shh and Gli-1 protein in each treatment group was significantly higher than that in the normal control group(P<0.05).The expression of Shh protein in the 7-day continuous treatment group was significantly higher than that in the model control group at the same time(P<0.05).Meanwhile,the expression of Gli-1 protein in the 7-day continuous treatment group was slightly higher than that in the 7-day model group,but P>0.05.The expression of Shh and Gli-1 protein in the 14-day continuous treatment group was significantly higher than that in the model control group at the same time period(P<0.05).Compared with the 14-day model group,the expression of Shh and Gli-1 protein in the acute treatment group was significantly increased,and P<0.05.The expression of Shh and Gli-1 protein in the recovery treatment group was also slightly higher than that in the model group,but P>0.05.The expression of Shh protein in acute treatment group was significantly higher than that in convalescent treatment group(P<0.05),but slightly lower than that in continuous treatment group for 14 days,P>0.05.The expression of Shh protein in rats treated for 14 days was significantly higher than that in rats treated for 7 days,P<0.05.At the same time,although the expression of Gli-1 protein in the 14-day continuous treatment group was higher than that in the convalescent treatment group,(P<0.05).However,although the expression of Gli-1 protein in acute treatment group is higher than that in recovery treatment group and lower than that in continuous treatment group for 14 days,there is no statistically significant difference(P>0.05).Conclusions:1.Warm acupuncture intervention can effectively improve behavioral indexes of spinal cord injury model rats,and warm acupuncture can promote recovery of limb motor nerve function after spinal cord injury.2.After spinal cord injury in rats,Shh and Gli-1 expression can be activated by itself to achieve slow repair of the body itself.3.Warm acupuncture can promote the expression of Shh and Gli-1 in Shh/Gli-1 signaling pathway,thus effectively promoting the proliferation and differentiation of neural stem cells,which may be an important mechanism for its treatment of spinal cord injury.4.Early intervention and continuous intervention of warm acupuncture after spinal cord injury can be more conducive to increasing mRNA and protein expression of Shh and Gli-1,and is more conducive to recovery of motor nerve function after spinal cord injury. |
PDF Full Text Request