| Objective Functional dyspepsia(FD)is a common gastrointestinal syndrome without organic lesions,and its etiology and pathological mechanism are complicated.Cajal Interstitial cells of Cajal(ICC)are pacemaker cells with slow-wave activity in the gastrointestinal tract,which can influence gastrointestinal motility by transmitting excitation and inhibitory signals.C-kit is a specific receptor for gastrointestinal ICC,also known as stem cell factor receptor.Stem cell factors(SCF)act as a series of signaling molecules activated by C-kit-specific ligands and C-kits through binding to the extracellular domains 1-3,thereby regulating ICC proliferation,differentiation,Development and structural integrity.Cx43 is a gap junction protein coexisting with C-Kit in ICC cells.Its expression has important pathophysiological significance for gastrointestinal dyskinesia.Decreased Cx43 expression can cause gastrointestinal motility disorders.From this,we speculate that mild moxibustion may change ICC's ultra-reticular structure and regulate the expression of C-kit,SCF,Cx43 protein and m RNA through SCF / C-kit pathway-mediated ICC proliferation and apoptosis.To improve the gastrointestinal dysfunction of FD and provide experimental evidence and new ideas for effective prevention and treatment of functional dyspepsia.Method Fifty healthy adult SD rats using SPF were randomly divided into 5 groups: blank control group,model group,mild moxibustion group,inhibitor group,mild moxibustion + inhibitor group,10 rats in each group.Except the blank control group,the other four groups were modeled by Guo's tail-clamp stimulation + irregular feeding+ ice-saline saline.After the model of the mild moxibustion group was successfully modeled,pure moxa was used to perform mild moxibustion at the Zusanli acupoints on both sides of the rats.Each moxibustion was performed continuously for 30 min.In the inhibitor group,on the 5th day after the modeling,the ACK2 group was given intraperitoneal injection of Kit neutralizing antibody(ACK2),0.2mg / Kg,for a total of 14 days.Moxibustion + inhibitor group provided moxibustion intervention on the basis of inhibitor group.After treatment,HE staining was used to observe the morphological and structural characteristics of gastric antrum tissue in each group;to observe changes in gastric emptying rate in each group;to observe the ultramicroscopic ICC in gastric antral tissue of each group by transmission electron microscope(TEM)Structural changes: Western blot was used to detect the protein expression of C-kit,SCF,Cx43 in gastric antrum tissue of each group;Real-time RT-PCR was used to detect the m RNA expression of C-kit,SCF,Cx43 in gastric antrum tissue of each group Variety.Result1.HE staining analysis: There was no ulcer,erosion,mesothelial cell reduction,and no obvious inflammatory cell infiltration or other organic lesions in the gastric antrum mucosa of each group of rats.2.Comparison of gastric emptying rate of rats in each group:Compared with the blank control group,the gastric emptying rate of model rats was significantly reduced and there was a significant difference(P <0.01),indicating that the FD rats were successfully constructed and FD rats appeared.Gastrointestinal motility disorders.Compared with the model group,the gastric emptying rate of rats in the mild moxibustion group was significantly increased,and the difference was statistically significant(P <0.05).The gastric emptying rate of the inhibitor group and the mild moxibustion + inhibitor group was compared with the model group.No significant difference(P> 0.05).3.Transmission electron microscope observation: In the blank control group,the interstitial cells of Cajal in the muscle showed spindle shape,dense cytoplasm,abundant endoplasmic reticulum mitochondria,and extended bipolar protrusions.Cajal interstitial cells in the model group were spindle-shaped and parallel to smooth muscle cells,but the protrusions were shortened and did not form a connection with nearby smooth muscle cells.In the mild moxibustion group,the shape of interstitial cells of Cajal changed,but the shape of the bipolar protrusion did not change,and it formed a gap connection with nearby nerves and smooth muscle cells.The interstitial cells of Cajal in the inhibitor group and the mild moxibustion + inhibitor group showed a spindle shape,the extension was longer than that in the model group,and a gap junction was formed.4.Comparison of protein expression levels of SCF,C-kit and Cx43 in gastric antrum tissue: Compared with the blank control group,the protein expression levels of SCF,C-kit and Cx43 in the model group were significantly reduced,and the difference was statistically significant(P <0.01).Compared with the model group,the protein expression levels of SCF,C-kit,and Cx43 in the mild moxibustion group were up-regulated,and the differences were statistically significant(P <0.05).The SCF,C-kit of the inhibitor group and the mild moxibustion + inhibitor group The expression levels of Cx43 and Cx43 were not significantly different from those of the model group(P> 0.05).5.Comparison of SCF,C-kit,and Cx43 m RNA in gastric antrum tissue: Compared with the blank control group,the m RNA of SCF,C-kit,and Cx43 in the model group showed low expression,and the difference was statistically significant(P <0.01);Compared with the model group,the m RNA expression levels of SCF,C-kit,and Cx43 in the mild moxibustion group increased(P <0.05),while the m RNA expressions of SCF,C-kit,and Cx43 in the mild and moxibustion+ inhibitor group were significantly higher than those in the mild moxibustion group.There was no significant difference between the model groups(P> 0.05).Conclusion1.This study found that mild moxibustion at Zusanli can significantly increase gastric emptying rate in FD rats,and this effect was reversed by the intervention of inhibitor ACK2,suggesting that mild moxibustion at Zusanli acupoints can improve gastric emptying and gastrointestinal tract in FD rats ICC is closely related.2.Mild moxibustion at Zusanli can increase the expression and transcription levels of SCF,C-kit and Cx43 in FD rats to a certain extent,increase the number of ICCs,improve the ultrastructure and function of ICC,and promote gastric motility recovery.3.The inhibitor ACK2 was used to specifically block the SCF/ C-kit signaling pathway.The protein and m RNA expressions of SCF,C-kit and Cx43 in the inhibitor group and the mild moxibustion +inhibitor group were not significantly different from the model group.Mild moxibustion The inhibitory effect of SCF,C-kit and Cx43 on the expression of SCF,C-kit and Cx43 was antagonized by the inhibitor,suggesting that mild moxibustion Zusanli can improve gastrointestinal motility of FD by regulating the SCF /C-kit signaling pathway. |
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