| Objective:To observe the effect of electroacupuncture at Shu and Mu acupoints in large intestine on the intestinal transit function of STC rats,the expression of GDNF in colon tissue and the methylation status of GDNF gene promoter region,and to treat the viscera disease with electroacupuncture Shu and Mu acupoints providing a scientific basis for the clinical application and mechanism research of acupoints.Methods: Sixty-four rats were randomly divided into 16 normal groups,16 saline groups,and 32 model rats.The STC model was replicated by compound diphenoxylate gavage method.The normal group was routinely fed and not treated.The saline group was administered with the same dose of normal saline.After the model was successfully established,the model was again randomly divided into 16 model groups and 16 electroacupuncture groups.The electroacupuncture group was treated with bilateral Tianshu and Dachangshu at the same time.The electroacupuncture treatment was performed for 15 minutes,once a day for 14 days.The rats in the remaining group were only fixed.The changes of behavior and defecation in each group were observed.The changes of intestinal transit function in each group were detected by black semi-solid paste propulsion method.The rat colon in different states was detected by Western blot and real-time fluorescent quantitative PCR.The expression of GDNF protein and mRNA was detected;the methylation level of the promoter region of GDNF gene was detected by BSP method.Result:1.Behavioral observation:(1)There was no significant difference in the scores of attack,modification and inquiry behavior between the normal group and the saline group(P>0.05).(2)Compared with the normal group and the saline group: the scores of attack,modification and inquiry behavior of the model group were significantly increased(P<0.01);the electroacupuncture group attack and inquiry behavior scores were statistically different(????);the modified behavior scores showed nosignificant difference(? ????))(3)Compared with the model group:electroacupuncture group modification,attack,and inquiry behavior score decreased(????).It is suggested that there is anxiety in STC rats.Electroacupuncture canimprove the attack,modification and inquiry behavior of STC rats,but the improvement of attack and inquiry behavior is worse than normal.2.Defecation:Comparison of 24-hour defecation weight and fecal trait scores in rats: After treatment,there was no significant difference between the normal group and the saline group(? ????);compared with the normal group and the salinegroup,the model group had a significant decrease in bowel movements(???),fecal trait score Reduced(????),there was no significant difference in theelectroacupuncture group(? ????);compared with the model group,theelectroacupuncture group showed a significant increase in the amount of defecation in24 hours(???),and the fecal trait score increased(????).It is suggestedthat the amount of defecation in STC rats is reduced,the feces are dry,and the electroacupuncture of the large intestines can promote defecation in rats and improve the symptoms of dry and hard stool texture.3.Comparison of intestine propulsion rate:There was no significant difference between the normal group and the saline group(P>0.05).Compared with the normal group and the saline group,the intestinal propulsion rate of the model group decreased significantly(P<0.01);The difference was statistically significant(P>0.05).The difference between the electroacupuncture group and the model group was statistically significant(P<0.01).It is suggested that the small intestine propulsion rate of STC rats is reduced,and the electroacupuncture of Shu and Mu acupoints in large intestine can increase the intestinal propulsion rate of STC rats,promote intestinal peristalsis and improve intestinal transit function.4.Expression of GDNF:GDNF protein and mRNA expression: There was no significant difference in the expression of histone and mRNA between the normal group and the saline group(P>0.05).Compared with the normal group and the saline group,the expression of histone and mRNA was significantly decreased(P><0.01);the expression of electroacupuncture group was increased compared with the modelgroup(P<0.05).It is suggested that the expression of GDNF protein and mRNA in colon tissue of STC rats is down-shifted,and the acupuncture of Shu and Mu acupoints in large intestine can increase the expression of GDNF protein and mRNA in colon tissue of STC rats.5.GDNF methylation level : There was no significant difference between the normal group and the saline group(P>0.05).Compared with the normal group and the saline group,the methylation level of the model group increased(P<0.05).Significance(P>0.05);the difference between the electroacupuncture group and the model group was statistically significant(P<0.05).It is suggested that the CpG methylation level of the GDNF gene promoter region in colon tissue in STC rats is increased,and the electroacupuncture of Shu and Mu acupoints in large intestine can reduce the methylation level.Saline administration had no significant effect on the results.Conclusion:1.Electroacupuncture Shu and Mu acupoints in large intestine can effectively improve constipation symptoms and intestinal transit function in STC rats.2.Electroacupuncture Shu and Mu acupoints in large intestine promotes the expression of GDNF by regulating the methylation status of the promoter region of GDNF gene,which may be one of the important mechanisms for the treatment of STC by electroacupuncture. |
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