| Objective: To observe the effects of 5-HT2 C antagonist on the symptoms,visceral sensitivity and the expression of 5-HT2 C,CGRP and SP in plasma,hippocampus,spinal cord and colon of IBS-D model rats regulated by herb-separated umbilical moxibustion,and to explore the mechanism of herb-separated umbilical moxibustion in the treatment of IBS-D from the perspective of brain-gut axis and 5-HT pathway.Methods: SPF SD healthy male rats were randomly divided into normal group,model group,umbilical moxibustion group and antagonism group,with 10 rats in each group.The normal group did not receive any treatment,and the other three groups were established into IBS-D model rats by acetic acid enema combined with chronic binding stimulation;The normal group and the model group did not receive treatment,but only grasping fixation and intraperitoneal normal saline injection synchronized with the antagonistic group;The umbilicus moxibustion group was treated with herb-separated umbilical moxibustion every three days,three times,and intraperitoneal normal saline injection synchronized with the antagonistic group was given at the same time;The antagonistic group was given intraperitoneal injection of 5-HT2 C receptor antagonist SB206553,1mg /kg once a day for 10 times,and was treated with herb-separated umbilical moxibustion as the umbilical moxibustion group at the same time.After treatment,the rats in each group were evaluated for 4h food and water intake,4h fecal points,body mass growth rate,fecal water content,fecal properties,abdominal withdrawal reflex(AWR)score,and then killed.The protein expression was detected by ELISA and Western blot,the protein localization expression was detected by immunohistochemistry,and the m RNA expression was detected by q RT-PCR.The data between multiple groups were analyzed by one-way ANOVA and expressed as mean ± standard deviation((?)±S).Result:1.Effects of herb-separated umbilical moxibustion and 5-HT2 C antagonist on symptoms and visceral sensitivity in IBS-D model ratsCompared with the normal group,the rats in the model group showed depression,dark hair color,significantly increased fecal water content,defecation times,fecal character score and AWR score(P < 0.01),and significantly decreased food intake and physical growth rate(P < 0.01);Compared with the model group,the umbilical moxibustion group significantly improved the above symptoms of IBS-D model rats(P <0.01);Compared with umbilical moxibustion group,the curative effect of antagonistic group on IBS-D model rats decreased significantly(P < 0.01).2.Effects of herb-separated umbilical moxibustion and 5-HT2 C antagonist on5-HT2 C,CGRP and SP protein in plasma,hippocampus,spinal cord and colon of IBS-D model ratsCompared with the model group,the umbilical moxibustion group significantly decreased the expression of 5-HT2 C,CGRP and SP protein in plasma,hippocampus,spinal cord and colon of IBS-D model rats(P < 0.01);Compared with the umbilical moxibustion group,the effect of down regulating 5-HT2 C,CGRP and SP protein in the antagonistic group decreased significantly(P < 0.01).3.Effects of medicine separated moxibustion on umbilicus and 5-HT2 C antagonist on5-HT2 C,CGRP and SP m RNA in hippocampus,spinal cord and colon of IBS-D model ratsCompared with the model group,the umbilical moxibustion group significantly decreased the expression of 5-HT2 C,CGRP and SP m RNA in plasma,hippocampus,spinal cord and colon of IBS-D model rats(P < 0.01);Compared with the umbilical moxibustion group,the effect of down regulating 5-HT2 C,CGRP and SP m RNA in the antagonistic group decreased significantly(P < 0.01).Conclusion:1.Herb-separated umbilical moxibustion may improve the symptoms of IBS-D model rats by reducing the activity of 5-HT pathway on brain-gut axis,and then down regulating the levels of CGRP,SP and other related brain gut peptides in 5-HT pathway..2.5-HT pathway may be one of the targets of medicine separated moxibustion on umbilicus in the treatment of IBS-D. |
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