| Background: Syndrome of deficiency of Spleen-Qi is one of the most common syndrome of deficiency in clinic,which with stomachache after eating,abdominal distension,fatigue,emaciation,loose stools,pale complexion,pale tongue,weak pulse and so on as the main clinical manifestations.Modern clinical research shows deficiency of spleen qi could take gastrointestinal motor dysfunction,digestion and absorption dysfunction,endocrine and immune dysfunction and many other changes as manifestations.The recovery of digestive tract dysfunction,digestive and absorption dysfunction,endocrine and immune function is based on the recovery of gastrointestinal motor function,the recovery of gastrointestinal motor function is the initial and most critical steps to correct the damage of gastrointestinal.Treatment of promoting the gastrointestinal motility can effectively improve the gastrointestinal motor dysfunction,digestion and absorption dysfunction,endocrine and immune function and other functions,which is a positive role in the process of treatment the deficiency of spleen-qi.Therefore,it is necessary to explore the nature of gastrointestinal motility dysfunction of spleen qi deficiency,to study the effective method of treating gastrointestinal motordysfunction of spleen qi deficiency.Acupuncture as a treatment Traditional Chinese Medicine,which has been applied for more than two thousand years in China.In clinical practice,more and more studies have shown that acupuncture at Zusanli/Tianshu acupoint can correct gastrointestinal motor function disorders,and treat the spleen-qi deficiency effectively,which has its own unique advantage in the treatment of functional gastrointestinal diseases.But the exact mechanism is still not very clear and requires further study.Objective: We established model rats of spleen-qi deficiency through the method of bitter diarrhea and qi loss.The ENS-ICC-SMC network of the small intestine of the spleen-qi deficiency syndrome was used to study the damage of ENS-ICC-SMC in the rats with spleen-qi deficiency syndrome,To study the pathogenesis of spleen deficiency syndrome.Then,the rats were treated with acupuncture at Zusanli and Tianshu acupoints to observe the effect of acupuncture on ENS-ICC-SMC network injury in rats with spleen-qi deficiency syndrome.To further study the mechanism of acupuncture at Zusanli and Tianshu acupoints.Methods: The adult Wistar rats were selected,both of the male and female.To establish the animal model of spleen qi deficiency syndrome by using physical screening,bitter diarrhea and qi consumption via referring to the method of Professor Chen Xiaoye of Chinese Academy of Traditional Chinese Medicine.The rats were divided into control group,model group,natural recovery group,false acupuncture group(acupuncture non-merdiain and non-acupoint group),treatment of acupuncture at Tianshu acupoint group,treatment of acupuncture at Zusanli acpoint group,treatment of acupuncture at Sanli+Tianshu acupoints combined group.After the success of the model establish and the treatment of the surviving rats,cervical dislocation after anesthesia,cut open the abdominal cavity,take the upper small intestine for the experiment.This study was divided into the following three parts: The first part,by observing the changes of general behavior,monitoring the changes of body weight,detecting the rate of D-xylose excretion in urine,observing the gross anatomy,detecting the rate of gastric emptying,detecting the rate of small intestinal advance,observing the HE staining pathology,detecting the ultrastructural structure by transmission electron microscopy,detecting immunofluorescence labelled subtance by laser scanning confocal microscopy and other methods to observe the morphological changes of ENS-ICC-SMC network in small intetine of rats with spleen qi deficiency,and the repairing effect of acupuncture at Zusanli and Tianshu acupoints.The second part,the method of detecting immunofluorescence double labelled substance by laser scanning confocal microscopy was used to observe the damage of the signal transduction pathway between the intestinal ENS-ICC of the spleen qi deficiency syndrome,and the repairing effect of acupuncture at Zusanli and Tianshu acupoints.The third part,by observing the injury of signal transduction pathway between ICC-SMC of small intestine in rats with spleen-qi deficiency via immunohistochemistry,Western blotting and qPCR,and the repairing effect of acupuncture at Zusanli and Tianshu acupoints.Through the three parts experiment,the essence of gastrointestinal motility dysfunction caused by spleen deficiency syndrome and the mechanism of acupuncture at Zusanli and Tianshu acupoints on spleen qi deficiency syndrome were discussed.Results:1.Compared with the normal control group,the general behavior of the rats in model group with syndrome of deficiency of spleen qi became bad evidently,the peristalsis of the small intestine by the general anatomical observation slowed down significantly,the structure of the small intestinal tissue by HE pathological findings was damaged,ICC ultrastructure was serious damaged,the gain of weight slowed down significantly,the rate of D-xylose excretion in urine,the rate of gastric emptying rate,the rate of small intestinal propulsion reduced significantly,the quantity of PGP9.5nerve fibers,ACh nerve fibers,SP nerve fibers,VIP nerve fibers,NO nerve fibers in the enteric nervous system,ICC,SMC and the network of ENS-ICC-SMC decreased significantly,the structure was serious damaged(P<0.05),but there was no significant difference between the model group,the natural recovery group and the false acupuncture group(P>0.05).Compared with the model group,there was a bit better in the Tianshu group and Zusanli group(P<0.05),but there were no significant differencesbetween the Tianshu group and the Zusanli group(P >0.05).Compared with Tianshu group and Zusanli group,the effect of the treatment of combined therapy group was obviously improved(P<0.05),but still lower than the normal control group(P<0.05).In general,the basic trend was: Model group ? Natural recovery group ? False acupuncture group <Tianshu group(ST25)? Zusanli group(ST36)< Combined group <Normal group.2.Compared with the normal control group,the signal transduction pathway between the enteric nerve-ICC of the rats in the model group with syndrome of deficiency of spleen qi,especially the quantity of the M receptors of the ACh nerve fibers,the NK receptors of the SP nerve fibers,the receptors of the NO nerve fibers –sGC,the VPAC receptors of the VIP nerve fibers,PKA,PKC and cGK/PKG were significantly decreased,and the signal pathways of G protein conjugated receptor-cAMP-PKA,G protein conjugated receptor-PLC-PKC,sGC-cGMP-PKG were serious destruction(P<0.05),but there was no significant difference between the model group,the natural recovery group and the false acupuncture group(P>0.05.Compared with the model group,there was a bit better in the Tianshu group and Zusanli group(P<0.05),but there were no significant differences between the Tianshu group and the Zusanli group(P>0.05).Compared with Tianshu group and Zusanli group,the effect of the treatment of combined therapy group was obviously improved(P<0.05),but still lower than the normal control group(P<0.05).In general,the basic trend was: Model group ? Natural recovery group ? False acupuncture group <Tianshu group(ST25)?Zusanli group(ST36)< Combined group < Normal group.3.Compared with the normal control group,the signal transduction pathway of ICC-SMC in the small intestine of the rats in model group with syndrome of deficiency of spleen qi,especially the quantity of the gap junction and connexin 43 were significantly reduced,the signal transduction pathway was serious destruction(P<0.05),but there was no significant difference between the model group,the natural recovery group and the false acupuncture group(P>0.05.Compared with the model group,there was a bit better in the Tianshu group and Zusanli group(P<0.05),but there were nosignificant differences between the Tianshu group and the Zusanli group(P>0.05).Compared with Tianshu group and Zusanli group,the effect of the treatment of combined therapy group was obviously improved(P<0.05),but still lower than the normal control group(P<0.05).In general,the basic trend was: Model group ? Natural recovery group ? False acupuncture group <Tianshu group(ST25)? Zusanli group(ST36)< Combined group < Normal group.Conclusion:1.With the spleen-qi deficiency syndrome,the general behavior of the rats in model group was the worst,the gain of weight was the slowest,the rate of D-xylose excretion in urine was the lowest,the peristalsis of the small intestine by the general anatomical observation was the slowest,the rate of gastric emptying rate,the rate of small intestinal propulsion were the lowest,the structure of the small intestinal tissue by HE pathological findings was damaged,ICC ultrastructure was serious destruction,the quantity of PGP9.5 nerve fibers,ACh nerve fibers,SP nerve fibers,VIP nerve fibers,NO nerve fibers in the enteric nervous system,ICC,SMC and the network of ENS-ICC-SMC were the least,the structure was serious destruction.The treatment of acupuncture at Zusanli and Tianshu acupoints could significantly repair the damage of spleen-qi deficiency.In general,the basic trend was: Model group ? Natural recovery group ? False acupuncture group <Tianshu group(ST25)? Zusanli group(ST36)<Combined group < Normal group.2.With the spleen-qi deficiency syndrome,the signal transduction pathway between the enteric nerve-ICC of the rats in the model group,especially the quantity of the M receptors of the ACh nerve fibers,the NK receptors of the SP nerve fibers,the receptors of the NO nerve fibers – sGC,the VPAC receptors of the VIP nerve fibers,PKA,PKC and cGK/PKG were significantly decreased,and the signal pathways of G protein conjugated receptor-cAMP-PKA,G protein conjugated receptor-PLC-PKC,sGC-cGMP-PKG were serious destruction.The treatment of acupuncture at Zusanli and Tianshu acupoints could significantly repair the damage of spleen-qi deficiency.In general,the basic trend was: Model group ? Natural recovery group ? Falseacupuncture group <Tianshu group(ST25)? Zusanli group(ST36)< Combined group <Normal group.3.With the spleen-qi deficiency syndrome,the signal transduction pathway of ICC-SMC in the small intestine of the rats in model group,especially the quantity of the gap junction and connexin 43 were significantly reduced,the signal transduction pathway was serious destruction.The treatment of acupuncture at Zusanli and Tianshu acupoints could significantly repair the damage of spleen-qi deficiency.In general,the basic trend was: Model group ? Natural recovery group ? False acupuncture group<Tianshu group(ST25)? Zusanli group(ST36)< Combined group < Normal group. |
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