| Objective:In this study,by establishing a rat model of LDH chronic lower extremity pain,selecting Duzhong Yaotong Pills as the target intervention drug,observing the analgesic effect of Duzhong Yaotong Pills on LDH chronic lower extremity pain model rats,and exploring Eucommia low back pain from the BDNF-Trkb-CREB signal channel The possible brain mechanism of the effect of pills in treating LDH chronic lower limb pain.Methods:1.Behavioral experiment: 72 male SD rats were randomly divided into normal group(Normal group n=24).(2)Model group(Model group n=24).(3)Model treatment group(Model Treatment group n=24).Autologous nucleus pulposus transplantation was used to model the model group and model treatment group respectively.After the model was successfully established,the normal group and the model group were given the same amount of normal saline by gavage,and the model treatment group was given Duzhong Yaotong Pill solution by gavage,and the course of medication was given.28 days.The rats in each group compared behavioral changes such as spontaneous academic score,mechanical pain threshold and thermal pain threshold at different time nodes.To observe the effect of Duzhong Yaotong Pill on the pain threshold of model rats.2.Molecular biology experiment: Based on the success of the behavioral experiment,4 rats in the normal group,model group,and model treatment group were randomly selected for brain hippocampal tissue 1 day before administration and 14 days and 28 days after administration.The materials were collected,and the expression levels of BDNF/Trkb/CREB protein and m RNA in the hippocampus of each group of rats were investigated by immunofluorescence,real-time fluorescence quantitative PCR,and western blotting.Results:1.The spontaneous pain behavior score results showed that the SD rats in each group performed well before the operation(P>0.05);after the operation model,compared with the normal group,the spontaneous pain behavior score of the model group and the model treatment group Significantly increased at 2 and 7 days after surgery(P<0.05),and at 14 days after surgery,the difference was not significant(P>0.05).After drug intervention in each group,compared with the model group,the model treatment group is being fed medicine Spontaneous pain behavior scores decreased significantly after 2days,7days,14 days,21days,and 28 days(P<0.05).2.The mechanical pain threshold results showed that: after the surgical model,compared with the normal group,the mechanical pain threshold of the model group and the model treatment group were significantly higher at 2 days and 7 days after surgery(P<0.05),and there was a difference at 14 days after surgery.Not significant(P>0.05).The overall observation of the model group and the model treatment group.The MWT of the model group showed an upward trend,indicating that the body gradually recovered after the successful model;the MWT of the model group stabilized after 21 days of feeding,and chronic pain was long-term Exist,but the MWT value of the model treatment group increased rapidly under the intervention of Duzhong Yaotong Pills(P<0.05).3.The results of thermal pain threshold showed that: after the surgical model,compared with the normal group,the thermal pain threshold of the model group and the model treatment group increased significantly at 1 and 7 days after surgery(P<0.05),and 14 days after surgery(At 0d feeding),the PWTL of the model group and the model treatment group increased to varying degrees,but the increase was small(P>0.05).After the drug treatment of Duzhong Yaotong Pills,compared with the model group,the model treatment group was feeding PWTL increased significantly after 2 days,7 days,14 days,21 days,and 28 days after treatment(P<0.05).4.IF test results show: After surgical modeling,compared with the normal group,the protein expression of BDNF protein and its specific receptors TrKB and CREB in the hippocampus of the model group was significantly inhibited 1 day before administration(14 days after modeling),with an average The fluorescence intensity was weakened,and the statistical data was comparable(P<0.01);after drug intervention,compared with the model group,the expression and distribution of BDNF,Trkb and CREB protein in the hippocampus of the model treatment group were 14 days after administration.Significant increase in 28 days,the average fluorescence intensity increased,and its statistical data is comparable(P<0.01).5.RT-qPCR results: Compared with the normal group,the expressions of BDNFm RNA,Trkbm RNA,and CREBm RNA in the hippocampus of the model group and the model treatment group were significantly reduced 1 day before the administration(14 days after the model was established)after the operation model was created(P <0.01);Compared with the model group,the expression of BDNFm RNA,Trkbm RNA,and CREBm RNA in the hippocampus of the model treatment group did not increase significantly 1 day before administration(P>0.01);after 14 days and 28 days of administration,the expression levels of BDNFm RNA,Trkbm RNA,and CREBm RNA in the hippocampus of the model treatment group did not increase significantly(P>0.01);Compared with the normal group,the expression levels of BDNFm RNA,Trkbm RNA,and CREBm RNA in the hippocampus of the model group were maintained at a lower level;compared with the model group,the expression levels of BDNFm RNA,Trkbm RNA,and CREBm RNA in the hippocampus of the model treatment group were in the administration 14 It increased significantly after 28 days and 28 days(P<0.01),and gradually approached the level of the normal group.6.WB test results show: Compared with the normal group,the BDNF,Trkb,and CREB protein expression levels of the model group and the model treatment group were significantly decreased after the model was established,that is,1 day before the administration(P<0.05);compared with the model group,the model treatment group was receiving The protein expression levels of BDNF,Trkb,and CREB did not change significantly one day before the drug(P>0.05);with the intervention of drugs and the extension of the intervention time,compared with the model group,the protein expression levels of BDNF,Trkb,and CREB in the model treatment group were There was a significant increase in 14 and 28 days after administration(P<0.05).Conclusion:Duzhong Yaotong Pill can effectively improve the pain behavior level of LDH chronic lower limb pain model rats,and promote the repair of nerve function damage caused by LDH.Duzhongyaotongwan up-regulates the expression levels of BDNF,Trk B and CREB protein and m RNA in the hippocampus of model rats.The mechanism of its effect on the treatment of chronic lower limb pain nerve injury in LDH may be related to the activation of the BDNF-Trk B-CREB signaling pathway in the hippocampus. |
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