| Objectives(1)To explore the therapeutic effect of electroacupuncture on defecation function of spinal cord injury on rats.(2)To clarify whether electroacupuncture improves the intestinal function of spinal cord injured rats through the mechanism of intestinal flora.Methods A total of 96 adult SPF grade SD female rats were selected.After adaptive feeding for one week,the rats were randomly divided into control group(sham,n=24)and spinal cord injury group(n=72).The model of severe spinal cord injury was established by means of weight drop,and only spinous processes and lamina were exposed in the sham group.All rats were intraperitoneally injected with gentamicin 2mg/kg to fight infection on the postoperative day,and the injury group was given postoperative nursing and urination once a day.After BBB score of 0 at 24 h,the injury group was randomly divided into 3 groups: model group(M,n=24),electroacupuncture group(E,n=24),and flora transplantation group(FMT,n=24).Only gentamicin was injected daily in the sham group and the M group,electroacupuncture was given to the E group(current intensity 1-2ma,continuous electrical stimulation for 15 min,once a day,14 days),and microflora was given to the FMT group(1ml/100 g body weight per day,10% microflora suspension was given to the FMT group,once a day,14 days,the microflora came from the fresh feces of group E).Intestinal transport function of rats in each group was recorded after 14 days of intervention: the volume of defecation(wet weight,dry weight and water content)for 24 hours,and the time of the first black feces,12 rats/group;Intestinal sensitivity test: AWR experiment,12 animals/group;Detection of bacterial diversity: 16 SrDNA sequencing and bioinformatics analysis,12 individuals/group;Pathological changes of colon were observed: HE staining and morphological observation,5 / group.Results 1.Establishment of rat model of severe spinal cord injury The BBB score of the model was 0 points 24 hours after the operation,which was higher than that of the sham group.The symptoms of fatigue were accompanied by urinary incontinence and urinary retention.Some of them were pyuria and hematuria.2.Effects of electroacupuncture and bacterial flora transplantation on intestinal transmission function In terms of wet weight,there was no significant difference between sham group and FMT group(p=0.07),and FMT group was significantly more than M group(p=0.046).In terms of water content,both group E and FMT were significantly higher than group M(both p<0.001).In the first case,both group E and FMT were significantly shorter than group M(both p<0.001),and group E was also significantly shorter than group sham(p=0.011).3.Effects of electroacupuncture and bacterial flora transplantation on intestinal sensory function There was no significant difference in the abdominal wall evacuation reflex scores among the four groups(all were p>0.05).4.Effects of electroacupuncture and microflora transplantation on pathological changes of colon After HE staining,the light microscope was enlarged by 100 and 200 times,and the results showed that,morphologically,the glands in group M were irregularly arranged,the muscle layer was thinner,and a large number of inflammatory cells were infiltrated.Similar to the FMT group,group E showed significant improvement in histopathology compared with group M,with more orderly arrangement of glands and more Numbers,thicker muscular layer,and only mild muscular layer atrophy and a small amount of inflammatory cell infiltration.5.Effects of electroacupuncture and bacterial flora transplantation on bacterial flora changes The results of 16 SrDNA showed that enough samples were taken in each group.Shannon index,Simpson index,Ace index and Chao1 index were used to compare intestinal microflora among each group.There were no significant differences in abundance and evenness in each group,all P>0.05.However,there were significant differences in the composition of bacterial colonies among all groups.Beta diversity index showed that there were significant differences in the bacterial colonies between SHAM group and E group,and sham and FMT groups(P<0.001);there were significant differences in the bacterial colonies between M group and E group,M group and FMT group(P<0.001),and no significant differences in other groups.The distribution of bacterial colonies among each group showed significant differences in the composition and structure of bacterial flora at the levels of phylum,class and genus:Electroacupuncture and microflora intervention can increase the abundance of melanobacteria at the portal level,reduce the abundance of proteobacteria and actinomycetes,and effectively reduce the proportion of-proteobacteria at the class level,but at the genus level,Electroacupuncture and fecal bacteria transplantation may affect intestinal function by further adjusting the abundance of Blautia,Dubosiella and Bacteroides,Bacteroides,Akkermansia and Lactobacillus.Conclusions(1)Electroacupuncture at zusanli acupoint can significantly improve the intestinal motor ability after spinal cord injury,causing benign morphological changes,and the morphological and behavioral results are consistent.(2)Electroacupuncture and its flora may improve intestinal transmission function by changing the composition of intestinal flora after spinal cord injury. |
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