推拿对坐骨神经慢性压迫性损伤大鼠背根神经节P2X_(3)受体表达及内向电流的影响

Effects of Tuina on P2X_(3) receptor expression and inward current in dorsal root ganglia of rats with chronic constriction injury of the sciatic nerve

目的:研究观察推拿对坐骨神经慢性压迫性损伤(CCI)大鼠背根神经节(DRG)P2X_(3)受体的表达以及通道电流变化的影响。方法:实验1:将32只SD大鼠随机分为空白组、CCI模型组、假手术组、CCI模型+推拿组,每组8只。推拿点按手法干预结束后,应用Western blot检测各组大鼠DRG神经元P2X_(3)受体表达情况。实验2:将12只SD大鼠随机分为空白组、CCI模型组、CCI模型+推拿组,每组4只,推拿点按手法干预结束后,应用全细胞膜片钳技术观察各组DRG神经元P2X_(3)通道内向电流的变化。结果:实验1:造模后,与空白组和假手术组比较,CCI模型组大鼠DRG中P2X_(3)的表达水平显著增高(P<0.01);CCI模型+推拿组P2X_(3)受体的表达水平显著低于CCI模型组(P<0.01),高于假手术组(P<0.05)。实验2:当给予α,β-meAPT浓度为0μL/L时,3组大鼠DRG P2X_(3)通道均未开放。当给予α,β-meAPT浓度为50μL/L时,3组大鼠DRG P2X_(3)通道均可诱发出不同程度的内向电流,但3组间电流幅度差异无统计学意义。当α,β-meAPT浓度为100μL/L时,CCI模型组DRG神经元P2X_(3)通道内向电流幅度较空白组显著升高(P<0.01),CCI模型+推拿组DRG神经元P2X_(3)通道内向电流幅度较CCI模型组显著下降(P<0.05)。结论:推拿可能通过抑制CCI模型DRG神经元P2X_(3)的表达以及通道开放程度,发挥镇痛作用。

Objective:To investigate the effects of Tuina on the expression of P2X_(3) receptors in dorsal root ganglia(DRG) and changes in channel current in rats with chronic constriction injury(CCI) of the sciatic nerve.Methods:Experiment 1:32 SD rats were randomly divided into blank,CCI model,sham,CCI model+Tuina groups,8 rats in each group.At the end of intervention by Tuina point according to manual technique,P2X_(3) receptor expression in DRG neurons of rats in each group was detected by Western blot analysis;Experiment 2:12 SD rats were randomly divided into blank group,CCI model group,CCI model+Tuina group,4 rats in each group.At the end of intervention by Tuina point in a manual manner,the whole cell patch clamp technique was applied to observe the changes of P2X_(3) channel inward current in DRG of blank group,CCI model group,CCI model+Tuina group.Results:Experiment 1:After modeling,the expression levels of P2X_(3) receptor in DRG of rats in the CCI model group were significantly higher compared with those in the blank and sham groups(P<0.01);The expression level of P2X_(3)receptor in the CCI model+Tuina group was significantly lower than that in the CCI model group(P<0.01) and higher than that in the sham group(P<0.05).Experiment 2:When given the concentration of α,β-meATP was 0 μL/L,P2X_(3) channels did not open in DRG of rats from all 3 groups.When the concentration of α,β-meATP was 50 μL/L,various degrees of inward currents were evoked through P2X_(3) channels in DRG of all three groups of rats,although there was no significant difference in current amplitude among the three groups.When the concentration of α,β-meATP was 100 μL/L,the inward current amplitude of P2X_(3) channels in DRG neurons in the CCI model group was significantly increased compared with that in the blank group(P<0.01),and P2X_(3)in DRG neurons in the CCI model+Tuina group.Channel inward current amplitude was significantly decreased compared with the CCI model group(P<0.05).Conclusion:Tuina may exert analgesic effects by inhibiting the expression of P2X_(3) as well as the degree of channel opening in DRG neurons of CCI model.