| Objective:The acupuncture of "Tiaoshen Changzhi" was use to interfere with PD model rats with constipation,to observe that effect of acupuncture of "Tiaoshen Changzhi" on gastrointestinal function of model rats,as well as on the structure of Interstitial cells of Cajal(ICC)in colon tissue and the expression of SCF and c-kit proteins,and to explore the possible regulatory mechanism of acupuncture of "Tiaoshen Changzhi" on PD with constipation.Implementation method:85 healthy male SD rats were selected and randomly divided into 12 Blank groups and12 Sham-operated groups.The other 61 rats were subcutaneously injected with rotenone suspension into the neck and back to establish the rat model of PD with constipation.The success of the model was determined by relevant behavioral evaluation and gastrointestinal function test.48 successful models were randomly divided into Model group,"Tiaoshen Changzhi" acupuncture group,Western-medicine group and Ordinary-acupuncture group.After completing the corresponding treatment intervention time of each group,the changes of gastric content residual rate,small intestine propulsion rate and fecal moisture content of rats in each group were observed.The relative expression of SCF and c-kit protein in colon tissue of experimental rats were detected by Western Blot and the structural changes of ICC were observed by electron microscope.The mechanism of acupuncture treatment with "Tiaoshen Changzhi" on PD with constipation model rats was discussed from the perspective of autophagy of Interstitial cells of Cajal and SCF/c-kit signal pathway.Research results:1.Fecal moisture content: before treatment,the fecal moisture content of the two acupuncture groups,Western-medicine group and Model group was significantly lower than that of the Blank group(P<0.01);after treatment,the fecal moisture content of rats in the Western-medicine group and the two acupuncture groups was higher than that in the Model group(P<0.01);Western-medicine group> "Tiaoshen Changzhi" group>Ordinary-acupuncture group.The difference between the three groups was statistically significant(P<0.01).2.Gastric content residual rate: after treatment,the gastric content residual rate of Model group was significantly higher than that of Blank group(P<0.01);Compared with the Model group,the gastric content residual rate in the Western-medicine group and the two acupuncture groups decreased significantly(P<0.01);Ordinary-acupuncture group > "Tiaoshen Changzhi" group >Western-medicine group.The difference between the three groups was statistically significant(P<0.01).3.Small intestine propulsion rate: after treatment,the small intestine propulsion rate of the Model group was significantly lower than that of the Blank group(P<0.01);Compared with the Model group,the small intestinal propulsion rate of rats in the Western-medicine group and the two acupuncture groups was significantly higher(P<0.01);Western-medicine group > "Tiaoshen Changzhi" group > Ordinary-acupuncture group.The difference between the three groups was statistically significant(P<0.01).4.Structural changes of ICC in colonic tissue: The ultrastructure of ICC in colonic tissue of rats was observed under a transmission electron microscope(multiple ×10).The results showed that the cells in the Model group had prominent surface,microvilli sparsely shortened,irregular nucleus,abundant organelles,and a large number of damaged mitochondrial cristae.There were a large number of vacuoles in the cytoplasm,enlarged intercellular space,severe swelling,and severe lymphocyte infiltration in the submucosa.In the Blank group,the cells had complete structure,no protrusions on the surface,regular nucleus,abundant organelles,normal intercellular space and no swelling.Compared with the Model group,the ICC structural damage in the colon tissue of rats in the Western-medicine group,"Tiaoshen Changzhi" group,and Odinary-acupuncture group was improved after treatment.In particular,the Western-medicine group had the most significant effect on the ICC structure repair of colon tissue,and the efficacy of the "Tiaoshen Changzhi" group was superior to that of the Odinary-acupuncture group.5.The relative expression of SCF protein in colon tissue was detected by Western Blot:the relative expression of SCF protein in colon tissue of Model group was significantly lower than that of Blank group(P<0.01),and there was no significant difference between Sham-operated group and Blank group(P>0.05).The relative expression of SCF protein in colon tissue of rats in Western-medicine group and two acupuncture groups was significantly higher than that in Model group(P<0.01).The effect of "Tiaoshen Changzhi" group was inferior to that of Western-medicine group(P<0.01),but better than that of Ordinaryacupuncture group(P<0.01).6.The relative expression of c-kit protein in colon tissue was detected by Western Blot:compared with the Blank group,the relative expression of c-kit protein in colon tissue of Model group was significantly lower(P<0.01),and there was no significant difference in the relative expression of c-kit protein in colon tissue of Sham-operated group compared with the Blank group(P>0.05).The relative expression of c-kit protein in colon tissue of rats in Western-medicine group and two acupuncture groups was significantly higher than that in Model group(P<0.01).The effect of "Tiaoshen Changzhi" group was inferior to that of Western-medicine group(P<0.05),but better than that of Ordinary-acupuncture group(P<0.01).Experimental conclusion:1."Tiaoshen Changzhi" acupuncture can better improve the gastrointestinal transmission function of PD with constipation model rats,reduce the gastric content residual rate,increase the fecal moisture content and the small intestine propulsion rate.2."Tiaoshen Changzhi" acupuncture can better repair the severe damage of ICC ultrastructure in PD with constipation model rats,and significantly increase the contents of SCF and c-kit protein in colon tissue.3.One of the mechanisms of "Tiaoshen Changzhi" acupuncture in the treatment of PD with constipation may be through the regulation of ICC and SCF/c-kit signal pathway. |
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