| Objective:1.The effect of abdominal massage on IBS-C rats was observed.2.To explore the effects of abdominal massage on EGCs,GDNF and AKt.Methods:1.The model of IBS-C rats was established by ice water gavage.The rats in the belly massage group,the model group and the blank group were separately raised in cages with normal feed and free water.Daily at 8:30 am,normal saline below 4?was used for intragastric infusion(3ml/only),continuous intragastric infusion for 14d,and the normal group was given the same amount of normal saline at room temperature.After the model was successfully established,rats in the tummy rub group were given tummy rub for 28 days,14min/d.Rats in the model group were only fixed for 14 minutes a day without any treatment for 28 days.The rats in the normal group were fed normally,without any treatment intervention,for a total of 28 days.The wet weight,dry weight and fecal particle number of rats were collected and calculated every day,and then the fecal character score and fecal water content were measured.Within 24 hours after the end of the modeling period and 24 hours after the end of the experimental period,intestinal sensitivity was measured in the three groups of rats.The volume thresholds of abdominal elevation and back arching were observed and recorded using the Abdominal evacuation reflex scoring(AWR)method.Each rat was measured for 3 times and the data were averaged.2.At the end of the measurement of test 1,rats were intraperitoneally anesthetized by injection of 2%pentobarbital sodium solution,then bled to death at bilateral common carotid artery,colon tissue was extracted,and PBS solution(PH=7.4)was immersed.At 4?,part of colon tissue was taken and fixed with 4%paraformaldehyde.Then colon tissue samples of each group were observed by transmission electron microscopy(TEM)×15000 and×30000.Image-pro6.0 Plus software was used to measure the ultrastructure of EGCs and calculate the area of heterochromatin and mitochondria of EGC cells.The expression of GDNF in colon of IBS-C rats and p-Akt in colon of IBS-C rats in each group were detected by Western blotting.Results:1.After the completion of modeling,the feces character score of rats in the rubbing group and model group was significantly lower than that in the blank group,and there was a statistical difference(~?P<0.05).Compared with the blank group,the number of feces particles in the model group and the rubbing group was statistically significant(P<0.05).The fecal water content of rats in the rubbing group and the model group was significantly reduced,which was significantly different from that in the blank group.Compared with the model group,the AWR threshold decreased significantly when the rectum balloon dilated(~?P<0.01).2.After abdominal rubbing in the experimental period,the feces traits score of rats in the abdominal rubbing group was close to that of the blank group,with significant difference compared with the model group(~?P<0.05).Compared with the model group,the number of feces particles of rats in the rubbing group was statistically significant(P<0.05).The fecal water content of rats in the rubbing group was close to that of the blank group.Compared with the model group rats,there were significant differences.The threshold value of AWR in rats caused by rectal balloon dilatation increased in the abdominal rubbing group,which was close to the blank group,and the difference was significant compared with the model group(~?P<0.01).3.Transmission electron microscopy showed that compared with the blank group,the heterochromatin area and the number of mitochondria in the model group decreased.Compared with the model group,the heterochromatin area was increased and the number of mitochondria was significantly decreased.4.Detection by Western blotting showed that GDNF protein expression was down-regulated compared with the blank group,and GDNF protein expression was significantly up-regulated after abdominal rubbing intervention.Further analysis of the PI3K/AKt signaling pathway found that compared with the blank group,phosphorylated AKt was significantly down-regulated in the model group,and phosphorylated AKt was significantly up-regulated after abdominal massage intervention.Conclusion:1.The dry and hard fecal matter,the number of fecal particles decreased,and the visceral sensitivity increased in rats in the rubbing group and the model group,which were statistically different from those in the normal group,proving that the model was successfully constructed by ice water gavage.2.In the experimental period,after abdominal rubbing intervention,the indexes of intestinal activity of rats in the abdominal rubbing group were significantly improved,which proved that abdominal rubbing method had a certain effect on IBS-C.3.Abdominal rubbing can adjust the ultrastructure of EGCs to improve THE symptoms of IBS-C.4.Abdominal rubbing can not only affect ENS by upregulating GDNF protein expression,but also change the signaling pathway of Akt,thus affecting IBS-C. |
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