| Purpose:The experimental study took the role of PVN in improving the gastrointestin al function of IBS-C rats with acupuncture and its mechanism as the starting poi nt,selected constipation-type irritable bowel syndrome animal models,and observ ed damaged PVN and electroacupuncture from the perspective of intestinal dyna mics The effects of four single points on the intestinal motility,visceral sensitivit y and the expression of CGRP and 5-HT in the serum and colon of IBS-C mod el rats.The expression of c-fos gene in the brain tissue of each group of rats w as observed to explore the effect of PVN.The role and mechanism of acupunctu re in improving the intestinal function of IBS-C rats.Method:(1)The IBS-C model was replicated using Cai Ying’s modified cold-ice-bind ing-starvation and satiety synthesis method.(2)The pain threshold of colorectal dilatation is used to represent the visceral sensitivity to evaluate the establishmen t of the IBS-C model.(3)The minimum volume threshold that can cause the rat’s abdominal retraction reflex is the pain threshold,and the effect of electroacupu ncture at the four single points and damaged PVN on the sensitivity of the rat’s viscera is evaluated.(4)Measure the intestinal propulsion rate and visceral sensi tivity of rats together as functional indicators to evaluate the effect of damaged PVN on the intestinal propulsion rate and visceral sensitivity of rats.(5)The ex pression of PVNc-fos protein in rats was measured by immunofluorescence techn ique.(6)Enzyme-linked immunosorbent assay was used to determine serum 5-H T and CGRP levels in rats.(7)The expression levels of 5-HT and CGRP in col on were detected by immunohistochemical method.Result:(1)(1)The results of visceral sensitivity showed that comparison to the nor mal control group,the minimum physiological saline water content,namely the minimum volume threshold of abdominal retraction reflex(AWR),which caused abdominal wall contraction reflex in model group was decreased(P<0.01),indicating that the visceral sensitivity of irritable bowel syndrome rats was increased.Th e model was copied successfully.Compared with model group,including four sing le cavity group’s abdomen of retraction reflex threshold(who runs AWR)minimu m capacity increased(P<0.01),PVN damaged-EA group who runs AWR mini mum capacity threshold no significant change(P>0.05),shows that acupuncture c an improve IBS-C rats state of visceral high sensitive,and damaged PVN after a cupuncture effect on the improvement of the visceral sensitivity is decreased obvi ously.(2)The measurement results of intestinal propulsion rate showed that,comp ared with the normal control group,the distance from the pylorus to the front of the black semi-solid pasting and the ratio of the pylorus to the ileocecum in m odel group were significantly decreased(P<0.01),indicating that the intestinal pr opulsion rate of IBS-C model rats was significantly decreased.Compared with the model group,the intestinal propulsing rate in the four single points group was significantly increased(P<0.01),while the intestinal propulsing rate in the damag ed group was not significantly changed(P>0.05),indicating that the intestinal pr opulsing rate in IBS-C rats could be significantly improved by the four single p oints electroacupuncture,while the effect of electroacupuncture on the improveme nt of intestinal propulsing rate was obviously weakened or even disappeared after PVN damage.(2)The immunofluorescence results of brain tissue showed that the nucleus was stained blue and the c-fos gene protein was stained green;compared with th e normal group,the expression of c-fos gene protein in the model group increas ed;compared with the model group,electroacupuncture IV The expression of c-f os gene protein decreased in single-hole group.(3)The enzyme-linked immunosorbent assay showed that the contents of 5-HT and CGRP in serum in model group were significantly higher than those in normal control group(P<0.01).Compared with the model group,the contents of 5-HT and CGRP in serum of the four single point electroacupuncture group were decreased(P<0.01).Compared with model group,serum 5-HT and CGRP content s in PVN-damaged EA group were not significantly different(P>0.05).These resu lts indicated that IBS-C could increase the contents of 5-HT and CGRP in seru m of rats,and the contents of 5-HT and CGRP in serum of IBS-C model rats were significantly decreased after electroacupuncture at four single points.However,after the damage of PVN,the regulation effect of acupuncture on the content s of 5-HT and CGRP in serum of IBS-C model rats was significantly weakened or even disappeared.(4)Colon immunohistochemical results showed that 5-HT and CGRP in the rat colon were mainly expressed in the mucosa and submucosa,the stained cells were brown and the nuclei were colorless.Compared with the normal group,th e positive expression rate of 5-HT and CGRP in the colon of the model group i ncreased significantly,and the difference was statistically significant.Compared w ith the model group,the positive rate of 5-HT and CGRP in the colon of the el ectroacupuncture four single-point group decreased,and the positive rate of 5-HT and CGRP in the colon of the PVN-EA group did not change significantly.It s hows that IBS-C increases the expression of 5-HT and CGRP in the colon tissue of rats.After electroacupuncture at four single points,the expression rate of 5-HT and CGRP in the colon tissue of IBS-C rats is regulated to close to the nor mal level,and it is damaged.After PVN,the effect of electroacupuncture on the four single points disappeared.Conclusion:(1)Electroacupuncture at four single points can reduce the expression of c-f os protein in PVN of brain tissue.(2)Electroacupuncture at four single acupoints can effectively increase the minimum volume threshold of AWR in IBS-C rats and improve the hypersensitiv ity state of IBS-C rats’ internal organs;at the same time,it can increase the inte stinal propulsion rate of IBS-C rats and promote intestinal motility.Electroacupu ncture was performed after the PVN was damaged.There was no statistically sig nificant change in intestinal propulsion rate and visceral sensitivity in IBS-C rats.(3)Electroacupuncture at four single acupoints can down-regulate the levels of 5-HT and CGRP in serum and colon of IBS-C rats,which may reduce viscer al sensitivity and improve intestinal function by reducing the levels of 5-HT and CGRP in serum and colon;After PVN is damaged,the down-regulation of 5-H T and CGRP levels in serum and colon of IBS-C rats by electroacupuncture disa ppears,indicating that electroacupuncture may use PVN as a channel to regulate5-HT and CGRP levels in serum and colon of IBS-C rats,And then have a the rapeutic effect on constipation-type irritable bowel syndrome. |
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