| Objective: By observing the expression of autophagy related factors LC3 and beclin-1 in substantia nigra and colon of rats with rotenone induced PD with gastrointestinal dysfunction,to explore the mechanism of the acupuncture method of ”regulating the spirit and activating the flow” on gastrointestinal dysfunction.Methods :Adult SD rats were randomly divided into blank group,sham operation group and model group.The rats in the sham operation group were intraperitoneally injected with 2.5ml/kg rotenone solution at the dose of 2.5ml/kg,5 days a week for 4weeks;the sham operation group was intraperitoneally injected with the same dose of1.25% chloroform and 4% sodium carboxymethyl cellulose solution,with the same cycle,and the blank group was not treated.Four weeks later,two rats were randomly selected from the blank group,two rats from the sham operation group,and eight rats were taken from the model group.After the determination,40 rats were divided into six groups according to the same method: tiaoxiangqing group,conventional acupuncture group,model group and Western medicine group,plus blank group and sham operation group,with 10 rats in each group.The western medicine group was given 75 mg / kg Madopar combined with 2 mg / kg domperidone by gavage;the sham operation group was given 5ml / kg normal saline;the mind regulating acupuncture group: the acupoints in Professor Wang Shun's "Tiao Shen Qing 36 9 acupuncture method" were selected,Baihui toutaiyang,Zhongwan,Qihai,Shenmen,Taichong and Sanyinjiao.Uniform use of0.25 mm × 25 mm filiform needle(and I brand),specific operation: Baihui penetrates the sun,the depth of acupuncture is 5mm,the lifting and inserting twisting range is large,the frequency is 200 times / min,and the needle is 9 times during the needle retaining period;the acupuncture depth of Zhongwan and Qihai points is 5mm,the lifting and twisting range is small,the frequency is 100 times / min,and the needle is6 times during the needle retaining period;the acupuncture depth of Shenmen,Sanyinjiao and Taichong is 3mm,only twisting without lifting and inserting,frequency is 3 mm The rate was 50 times / min.during the period of retaining needle,the needle was kept for 30 minutes for 3 times.Once a day for 2 weeks.Acupuncturewas performed by the same person;in the conventional acupuncture group,the "dance tremor control area" and "Tianshu","Zusanli" and "Shangjuxu" points were selected,with a depth of 5mm,and conventional acupuncture was performed for intervention treatment;each group was treated once a day for 2 weeks.After 2 weeks of treatment,the expression levels of autophagy related factors LC3 and Beclin1 in substantia nigra and colon were observed by immunohistochemistry and protein imprinting.Results:1.The expression of LC3 and beclin-1 in the substantia nigra of the rat midbrain was observed by immunohistochemistry and protein imprinting.(1)Compared with the normal group,the content of LC3 and Beclin1 in the substantia nigra of the model group decreased significantly(P < 0.01),which was statistically significant.(2)Compared with the model group,the content of LC3 and Beclin1 in the substantia nigra of the middle brain of the rats in the mood group increased significantly(P <0.01),which was statistically significant.(3)Compared with the model group,the contents of LC3 and Beclin1 in the substantia nigra of the midbrain in the conventional acupuncture group were significantly higher(P < 0.01).(4)Compared with the model group,the content of LC3 and Beclin1 in the substantia nigra of the middle brain in the western medicine group increased slightly(P < 0.01),which was statistically significant.2.The expression of LC3 and beclin-1 in colon of rats was observed by immunohistochemistry and protein imprinting.(1)Compared with the normal group,the LC3 and Beclin1 contents in the colon tissue of the model group were significantly higher(P < 0.01).(2)Compared with the model group,the content of LC3 and Beclin1 in the colon tissue of the rats in the mood group decreased significantly(P < 0.01),which was statistically significant.(3)Compared with the model group,the contents of LC3 and Beclin1 in the colon tissue of rats in the conventional acupuncture group were significantly reduced(P<0.01).(4)Compared with the model group,the content of LC3 and Beclin1 in colon tissue of the western medicine group was slightly lower(P < 0.01),which was statistically significant.Conclusion:1.Western medicine has the most obvious effect in improving behavioral score,fecal water content and solid food residue rate in stomach of PD rats with gastrointestinal dysfunction,followed by "Tiao Shen Qing" acupuncture therapy,and "Tiao Shen Qing" acupuncture therapy is superior to conventional acupuncture treatment.2.The results showed that the contents of LC3 and Beclin1 in brain tissue were significantly decreased in PD rats with gastrointestinal dysfunction,while the contents of LC3 and Beclin1 in colon tissue were significantly increased.3."Tiaoshenqing" acupuncture treatment can significantly increase the content of LC3 and Beclin1 in brain tissue of PD gastrointestinal dysfunction model rats,and it is better than conventional acupuncture treatment;it can significantly reduce the content of LC3 and Beclin1 in colon tissue,which is better than conventional acupuncture treatment.4.The treatment of PD gastrointestinal dysfunction by "Tiao Shen Qing Qing" acupuncture may be achieved by regulating the content of autophagy related factors LC3 and Beclin1 in different parts. |
PDF Full Text Request