Mechanism of Tuina in rats with neuropathic pain based via autophagy and apoptosis of dorsal horn of spinal cord
Objective It is to explore the effect of Tuina on autophagy and apoptosis of spinal cord dorsal horn in rats with neuropathic pain,and to reveal its mechanism of inhibiting inflammation and analgesia.Methods Thirty SD rats were randomly divided into 5 groups with 6 rats in each group:including normal group,sham operation group,model group,Tuina group and sham Tuina group.Neuropathic pain models were established by ligating the L5 spinal nerve in the rats of all groups except for the normal group and sham operation groups.Starting on the 1st postoperative day,the rats of the Tuina group were stimulated at their acupoints of Huantiao,Yanglingquan and Xuanzhong on the bile meridian by pointing,dialing and kneading sequentially for a total of 18 min,and the hind limbs of rats of the sham Tuina group were gently stroked for 18 min,all once daily,continuously treated for 14 days;the rats of the rest groups were not treated,and were observed for 14 days.The mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)of rats in each group were measured before surgery and on the 1st,3rd,7th and 14th postoperative days;at the end of the treatment on the 14th postoperative day,the serum level of tumor necrosis factor-α(TNF-α)was detected by ELISA,and the expressions of Beclin1,LC3Ⅱ,Bcl-2,Caspase-3 proteins in L4-6 spinal cord dorsal horn tissues were detected by Western blot,and the expressions of Beclin1 and Bcl-2 mRNA in L4-6 spinal cord dorsal horn tissues were detected by RT-qPCR.Results On the 1st,3rd postoperative days,there was no significant difference in MWT and TWL of rats among the model group,Tuina group and sham Tuina group(all P＞0.05),but the MWTs were significantly lower,while the TWLs were significantly shorter than those at the same period in the normal group and sham operation group(all P＜0.05).On the 7th and 14th postoperative days,the MWTs of rats in the Tuina group were significantly higher,while their TWLs were significantly lon-ger than those at the same period in the model group and sham Tuina group(all P＜0.05).Compared with the normal group and sham operation group,the serum level of TNF-α and the relative expression of Caspase-3 protein in spinal cord tissues in the model group were significantly higher(all P＜0.05),and the relative expressions of Beclin1,LC3Ⅱ,Bcl-2 proteins and the relative expressions of Beclin1 and Bcl-2 mRNA in spinal cord tissues were significantly lower(all P＜0.05);compared with the model group,the serum level of TNF-α and the relative expression of Caspase-3 protein in spinal cord tissues in the Tuina group were significantly lower(all P＜0.05),and the relative expressions of Beclin1,LC3Ⅱ,Bcl-2 proteins and Beclin1 and Bcl-2 mRNA in spinal cord tissues were significantly higher(all P＜0.05);compared with the sham Tuina group,the serum level of TNF-α and the relative expression of Caspase-3 protein in spinal cord tissues in the Tuina group were significantly lower(all P＜0.05),and the relative expressions of Beclin1,Bcl-2 proteins and Bec-lin1 mRNA in spinal cord tissues were significantly higher(all P＜0.05),the difference in the relative expression of Bcl-2 mRNA in spinal cord tissues was not significant between the two groups(P＞0.05).Conclusion Tuina could significant-ly alleviate the pain response in rats with neuropathic pain,and its mechanism may be related to up-regulating the expres-sion of autophagy-related gene Beclin-1 and anti-apoptotic factor Bcl-2 in dorsal horn of spinal cord,which enhances auto-phagy and inhibit apoptosis,and attenuates the immune-inflammatory response mediated by pro-inflammatory factors.
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万方数据
维普
