Study On Effects Of Electroacupuncture At Back-shu And Front-mu Points On GDNF-PI3K-Akt Signal Transduction Pathway In GFRα1-knockout And Functional Constipation Mice

Objective:To explore the mechanism of electroacupuncture(EA)at Back-shu and Front-mu points on GDNF-PI3K-Akt signal pathway in functional constipation(FC),based on the efficacy of acupuncture in the treatment of functional constipation and the guidance of previous results,so as to provide scientific basis for acupuncture Back-shu and Front-mu points for the treatment of functional constipation,in order to facilitate the clinical application.Methods:10 wild-type mice were set as control group,and 13 GFRα1 knockout mice were randomly assigned to two groups:Model control(n =7),acupuncture group(n = 6).We established the model by compound diphenoxylate orally.The animals underwent electroacupuncture(EA)treatment at Tianshu(ST 25),Dachangshu(BL 25)on each side about alternate acupuncture for 30 minutes,once a day,a total of five treatments.The Gastric emptying,The small intestine transmitting rate,were examined after the treatment.We examined the GDNF-PI3K-Akt after treatment by Digital scanning microscope,Electron microscope,TUNEL,Immunohistochemistry,in situ hybridization,PCR and Western Blot in the distal colon.Results:1、Gastrointestinal Transmitting Function:(1)Gastric emptying rateCompared with the control group,the gastric emptying rate in the model group and the acupuncture group was decreased(P<0.01,P<0.01);compared with the model group,the gastric emptying rate was not significantly changed in the acupuncture group(P>0.05).(2)Small intestine transmitting rateCompared with the control group,the small intestine transmitting rate in model group was decelerated(P<0.05).and the acupuncture group also decreased(P<0.01).In the acupuncture group,the effect of acupuncture on small intestine transmitting rate was not obvious compared with the model group(P>0.05).2、GDNF-PI3K-Akt signal pathway(1)The expression of GDNF protein and mRNACompared with the control group,the expression of GDNF protein and mRNA in the colon of the model group had a decreasing trend,but there was no statistical significance(P>0.05,P>0.05).In the acupuncture group,the GDNF protein and mRNA had no difference compared with the model group(P>0.05,P>0.05).(2)The expression of Rai protein and mRNAIn model group,the content of Rai protein and mRNA was significantly reduced than those in control group(P<0.01,P<0.01).In the acupuncture group,the GDNF protein and mRNAhad no difference compared with the model group(P>0.05,P>0.05).(3)The expression of RET protein and mRNAIn model group,the content of RET protein was significantly reduced than those in control group(P<0.01),the mRNA has no statistically significant(P>0.05).In the acupuncture group,the GDNF protein and mRNA had no difference compared with the model group(P>0.05,P>0.05).(4)The expression of PI3 K and AktIn model group,the content of PI3 K and Akt was significantlyincreased than that of control group(P<0.01,P<0.01).In acupuncture group,the level of PI3 K and Akt was decreased(P<0.05,P<0.05).Conclusions:1、After GFRα1 knockout,acupuncture at the Back-shu and Front-mu points has no obvious effect on the improvement of gastrointestinal transport function in mice withfunctional constipation.2、After GFRα1 knockout,acupuncture at the Back-shu and Front-mu points has no obvious effect on the expression level of each molecule of GDNF-PI3K-Akt signaling pathway in functional constipation mice.From the reverse side,it was confirmed that GDNF-PI3K-Akt signal transduction pathway may be an important path for functional constipation with acupuncture at the Back-shu and Front-mu points.3、GDNF is not the only molecule that mediates the PI3K-Akt signaling pathway.The effect of acupuncture on the expression of PI3K-Akt signaling pathway may include other ways.