基于GAS、NO和MLCK信号通路观察一指禅推法干预脾虚家兔的作用机制

Action Mechanism of One-Finger Pushing Intervention in Rabbits with Spleen Deficiency Based on GAS,NO and MLCK Signal Pathways

目的:研究一指禅推法对脾虚家兔GAS及NO影响,且通过抑制MLCK信号通路,观察一指禅推法干预脾虚家兔的作用机制。方法:取成年新西兰兔40只,随机分为空白组(K组)、模型组(P组)、一指禅推法治疗组(A组)、抑制剂组(I组)。除K组外,P组、A组及I组进行造模。A组与I组使用人工机械臂于中脘穴、天枢穴(双)行一指禅推法,频率为101~150次/min,压力为150mN,其中I组的家兔模型在每日手法治疗后均进行腹腔注射ML-9(MLCK强抑制剂)。治疗10d后处死,取血清样本测量胃泌素(GAS),取胃窦组织样本测量一氧化氮(NO)水平,且进行治疗后各组家兔进食量的对比。结果:①GAS水平研究发现,K组(7.17pg/mL)水平最低,P组(78.82pg/mL)水平最高,A组(20.66pg/mL)和I组GAS(19.96pg/mL)水平仍无法降至K组的水平,但较P组已有明显下降,而两组之间差异无统计学意义(P>0.05);②NO水平研究发现,K组水平最高(46.64μmol/g),P组水平最低(4.03μmol/g),A组(31.02μmol/g)和I组(31.00μmol/g)水平较K组下降,较P组上升,而两组之间差异无统计学意义(P>0.05);③进食量对比研究发现,治疗后,A组及I组进食量较P组进食量升高,而尚未达到K组水平。综上,A组和I组在GAS、NO和家兔治疗后的进食量等指标中并无明显差异,说明一指禅推法并非仅经MLCK通路对脾虚家兔起干预作用。结论:一指禅推法可通过多通路、多途径影响脾虚型家兔的GAS、NO水平,从而促进脾虚型家兔的胃肠功能趋于正常水平。

Objective:To study the effect of one-finger pushing on GAS and NO in the rabbits with Spleen deficiency,and to explore the action mechanism of the therapy by inhibiting MLCK signal pathway.Methods:40 rabbits were randomly divided into the blank control group(group K),the model group(group P),the one-finger pushing group(group A)and the inhibitor group(group I).After successful modeling,group A and group I were given one-finger pushing on bilateral Zhongwan(RN12)and Tianshu(ST25)with frequency of 101-150 times/minute as well as pressure of 150mN by robotic arm.Group I was given intraperitoneal injection of ML-9 after the routine treatment.After 10 days of the intervention,the serum level of gastrin(GAS)and the content of nitric oxide(NO)in gastric antrum tissue were detected,and the rabbits in each group were compared in terms of food intake after the treatment.Results:In terms of GAS level,it was lowest in group K,which was 7.17 pg/mL;it was highest in group P,which was 78.82 pg/mL;it was 20.66 pg/mL in group A and 19.96 pg/mL in group I,which were lower than that in group P,but higher than that in group K,and there was no statistical difference between group A and group I(P>0.05).In terms of NO level,it was highest in group K,which was 46.64μmol/g;it was lowest in group P,which was 4.03μmol/g;it was 31.02μmol/g in group A and 31.00μmol/g in group I,which were lower than that in group K but higher than that in group P;there was no statistical difference between group A and group I(P>0.05).In terms of food intake change,it was higher in group A and group I than that in group P,but did not reach the level of group K;there was no significant difference in GAS,NO and food intake between group A and group I after the treatment,indicating that one-finger pushing did not only intervene the rabbits with Spleen deficiency through MLCK pathway.Conclusion:One-finger pushing therapy can affect the levels of GAS and NO in the rabbits with Spleen deficiency through multiple pathways and multiple approaches,so as to promote the recovery of gastrointestinal function in the rabbits with Spleen deficiency.