| Objective: To investigate the effect and mechanism of electroacupun-cture combined with acupoint catgut embedding ultra-advanced intervention in post-operative anal pain model rats.Methods: Thirty healthy and clean SD rats were selected.After excluding the rats with abnormal pain threshold,the rats were randomly divided into 5groups according to body weight,with 6 rats in each group,which were blank control group(A),model group(B),electro-acupuncture group(C),acupoint catgut embedding group(D)and electro-acupuncture combined with acupoint catgut embedding group(E).Group A and group B received no treatment intervention,but group B received the preparation of postoperative anal pain model.Group C was given electro-acupuncture to stimulate Zusanli and Changqiang points 30 min before modeling;In group D,sheep intestinal thread was buried in Zusanli and Changqiang points 30 minutes before modeling;Group E was treated with electro-acupuncture 30 minutes before modeling according to the operation method of group C,and then treated with acupoint catgut embedding according to the operation method of group D.The mental state,diet,water,body weight,biting behavior and local anal incision of rats in each group were observed,and the volume of defecation 6h after the model was recorded.Mechanical pain threshold at anal incision was measured by Von Frey needle on day 1(T0)before intervention and 2h(T1),12h(T2),24h(T3),day 3(T4)and day 7(T5)after modeling,respectively.After mechanical threshold measurement,abdominal aorta blood and local anal incision tissue were taken under anesthesia with 2% pentobarbital sodium(40 mg/Kg),and the incision part of anus was observed by histomorphology.The contents of PGE2,TNF-α,IL-1β and BDNF in serum and local anal tissue of rats were detected by ELISA.The expression levels of Wnt-1,β-catenin and GSK-3β in local anal tissues of rats were detected by Western blot.Results: 1.General results:(1)General information: rats in group A had good mental state,normal diet and water intake,no anal redness,anal incontinence,etc.The mental state of rats in group B was poor,the amount of food and water was significantly reduced,the perianal incision was redness and swelling,there was anal biting behavior,no anal incontinence;The mental state of the rats in groups C,D and E was poor after modeling,and the amount of food and water consumption was significantly reduced.However,the mental state and the amount of food and water consumption of the rats in groups C and D basically returned to normal on the 7th day after modeling,with slight redness and swelling of the anus,incision healing and scab,no biting behavior of the anus,and no anal incontinence.On the 7th day after modeling,the mental state,food and water intake of rats in group E completely returned to normal,and there was no redness,swelling,healing and scab of anal incision,anal biting behavior,or anal incontinence.(2)Body mass: there was no statistical significance in body mass before and after the experiment and between groups(p > 0.05).(3)Defecation volume 6h after modeling: compared with group A,the defecation volume in group B was significantly decreased within 6h after modeling,and the difference was statistically significant(p < 0.05).Compared with group B,the defecation volume in group C,D and E was significantly increased within 6h after modeling(p < 0.05).Compared with group E,the defecation volume of rats in group C and D was significantly reduced within 6h after modeling(p < 0.05),but the difference between groups C and D was not significant,the difference was not statistically significant(p > 0.05).2.Mechanical pain thresholds: there was no significant difference in basal pain threshold(T0)among all groups(p > 0.05).Compared with group A,the mechanical pain threshold of group B,C,D and E was significantly decreased after modeling,with statistical significance(p < 0.05).Compared with group B,the mechanical pain threshold of group C,D and E was significantly increased after modeling(p < 0.05),and the increasing trend and value of the mechanical pain threshold of group E were the most obvious;Before T3 time point,the mechanical pain threshold of group C was higher than that of group D(p <0.05),and after T3 time point,the increase trend of mechanical pain threshold in group D was more obvious than that in group C(p < 0.05).At T5 time point,the mechanical pain threshold of groups C and D was significantly lower than that of group E(p < 0.05),but the difference was not statistically significant compared with groups C and D(p > 0.05).3.Histopathological observation: the muscle fibers in group A were intact with normal shape and no obvious inflammatory cell infiltration.In group B,there were different degrees of necrosis and atrophy of muscle fibers,a large number of inflammatory cells mainly lymphocytes could be seen,and occasionally fibroblast hyperplasia in the interstitial muscle.A small number of inflammatory cell infiltration,slight edema,small number of new capillaries and fibroblast proliferation were observed in group C and D.In group E,there was no obvious inflammatory cell infiltration,edema disappeared obviously,and a large number of new capillaries and fibroblast hyperplasia could be seen in the interstitium.4.Contents of PGE2,BDNF,TNF-α and IL-1β:compared with group A,the contents of inflammatory mediators PGE2,BDNF,TNF-α and IL-1β in serum and tissues of groups B,C,D and E were significantly increased,and the differences were statistically significant(p< 0.05).Compared with group B,the expression level of group C,D and E was gradually decreased as time went on,and the expression level of group E was significantly lower than that of groups C and D on the 7th day(p <0.05),but there was no statistical difference in the inflammatory indicators among groups C and D(p > 0.05).5.Expression levels of Wnt-1,β-catenin and GSK-3β : compared with group A,the protein expression levels of Wnt-1 andβ-catenin in groups B,C,D and E were significantly increased,while the protein expression levels of GSK-3β were significantly decreased,with statistical significance(p < 0.01).As time went on,compared with group B,the protein expression levels of Wnt-1 and β-catenin in groups C,D and E were significantly decreased(p < 0.01),while the protein expression level of GSK-3β was significantly increased(p < 0.01).The protein expression levels of Wnt-1 and β-catenin in group E on the 7th day were significantly lower than those in groups C and D(p < 0.01),and the protein expression levels of GSK-3β in group E were significantly higher than those in groups C and D(p <0.01),but there was no statistical difference in the protein expression levels between groups C and D(p > 0.05).Conclusion: 1.Electro-acupuncture combined with acupoint catgut embedding advanced intervention has a fast and lasting effect on postoperative anal pain in rats,and has a better effect than single use of electro-acupuncture or acupoint catgut embedding.2.There is no significant difference in analgesic effect between single use of electro-acupuncture or acupoint catgut embedding therapy,but electro-acupuncture therapy has quick effect and short duration of effect.The effect of acupoint catgut embedding treatment was slow,but the effect lasted for a long time.3.Electro-acupuncture combined with acupoint catgut embedding ultra-advanced intervention can play a synergistic role in significantly alleviating postoperative anal pain,reducing inflammatory response,improving defecation and accelerating incision healing in rats,which may be related to the inhibition of Wnt/β-catenin signaling pathway.4.Electro-acupuncture combined with acupoint catgut embedding advanced intervention of acupoint embedding has a positive effect on the treatment of postoperative anal pain in rats,and is worth popularizing clinically. |
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