| Objective: In this study,diabetic gastroparesis model rats were selected to investigate the effect of acupoint selection on gastric movement.In addition,the possible mechanism of the effect difference should be discussed from the different perspectives of molecular biology and brain metabolism,so as to provide scientific basis for the research of material basis and function of acupoint selection according to location.Methods: In this study,Zucker rats were fed with high-fat diet for 16 weeks to produce diabetic gastroparesis model.The rats were divided into Normal group,Model group,Domperidone group,CV12 group,BL21 group,PC6 group and ST36 group according to the different acupoint selection sites.The effect of different acupoints on gastric movement in DGP model was evaluated by SPECT/CT.Systemic morphology,molecular biology and PET/CT related experimental techniques were used to observe the effects of acupuncture on the morphology of gastric antrum,the ultrastructure of ICCs,proteins related to SCF/c-kit signaling pathway,serum brain-gut peptides and glucose metabolism in different brain regions in DGP model rats.Results:1.The blood glucose and gastric emptying results: compared with Model group,Domperidone group had significant difference(P<0.001),CV12 group had significant difference(P<0.05),BL21 group had significant difference(P<0.01),PC6 group had significant difference(P<0.001),ST36 group had significant difference(P<0.01).There was no significant difference in blood glucose between groups(p>0.05).When gastric emptying for 30 minutes,there was significant difference between Model group and Normal group(P<0.01).At 40 min,there was a significant difference between Model group and Normal group(P<0.001),and a significant difference between ST36 group and Model group(P<0.05).At 50 min,there was a significant difference between Model group and Normal group(P<0.001),and a significant difference between ST36 group and Model group(P<0.01).At 60 min,there were significant differences between Model group and Normal group(P<0.001),Domperidone group and Model group(P<0.05),PC6 group and Model group(P<0.05),and ST36 group and Model group(P<0.01).2.The results of hematoxylin-eosin staining in gastric antrum: In Normal group,the SMC of the gastric antrum were arranged orderly and closely;the glandular cells of the gastric antrum mucosa and submucosa were arranged regularly,and the intercellular space was moderate,without vacuolation and vasodilation.In Model group,the cytoplasm of the SMC in the antrum was light and vacuolated;the glandular cells in the antrum mucosa and submucosa were not arranged orderly,the intercellular space became larger,the number of main cells decreased,the cytoplasm of the wall cells decreased,vacuolated and stained light,and the blood vessels were dilated.Compared with Model group,Domperidone group and other four acupuncture groups showed that the arrangement of SMC in the muscular layer of gastric antrum was relatively dense;the arrangement of glandular cells in the mucosal layer and submucosa was relatively orderly,the number of main cells was relatively increased,the cell gap was reduced,the vacuole was reduced,and the congestion was reduced.3.Ultrastructural results of ICCs in antrum of stomach under electron microscope: In Normal group,the ultrastructure of ICCs in gastric antrum was clear,large in number and volume,and the cell morphology was basically normal.The nucleus is normal,the structure is clear,the heterochromatin is rich,and the nucleolus is obvious;the organelle structure in the cytoplasm is normal,and the mitochondria are very rich,closely arranged in the cytoplasm.It is closely connected with adjacent cells and peripheral nerve fiber endings.Compared with Normal group,the number of ICCs in the antrum of Model group was significantly reduced,and the volume of ICCs was reduced,and the cell morphology and structure were abnormal.There are a lot of vacuoles and organelles,swelling and vacuolation of mitochondria,disintegration of nucleolus and marginalization of heterochromatin,and partial destruction of intercellular connections.The connection with adjacent cells and peripheral nerve fiber terminals is loose,and the reticular structure is damaged in varying degrees.The number of ICCs in antrum of Domperidone group was higher than that of Model group.Some cells were abnormal.There are autophagy bodies and secondary lysosomes in the cells;there are abundant mitochondria in the cytoplasm,and the morphology is basically normal,but some of them have abnormal arrangement and swelling of cristae;there are abundant heterochromatism in the nucleus.The connection with adjacent cells and peripheral nerve fiber terminals was still loose in varying degrees,but it was improved compared with the model group.In the overall experimental group,the shape of Domperidone group was slightly better than that of the other four acupuncture groups,all of which were better than that of the model group,but there were still a lot of abnormalities.4.SCF/c-kit signaling pathway(1)The results of immunohistochemical in gastric antrum with SCF: compared with Normal group,the expression of Model group was decreased.Compared with Model group,the expression of Domperidone group,CV12 group,BL21 group,PC6 group and ST36 group were all increased to varying degrees.(2)The results of immunohistochemical in gastric antrum with c-kit: The expression in Model group was lower than that in Normal group.Compared with Model group,the expression of Domperidone group,CV12 group,BL21 group,PC6 group and ST36 group increased in varying degrees.(3)The results of RT-PCR: Compared with Normal group,the content of SCF mRNA and c-kit mRNA in antrum of Model group was lower than that of Normal group,and the difference was statistically significant(P<0.001);Compared with Model group,the SCF mRNA content in Domperidone group and four acupuncture groups was increased,only the increase in BL21 group,PC6 group and ST36 group was statistically significant(P<0.01,P<0.001);Compared with Model group,the content of c-kit mRNA in Domperidone group and four acupuncture groups was increased,only the increase in Domperidone group was statistically significant(P<0.05).(4)The results of ELISA: The serum SCF level in Model group was significantly lower than that in Normal group(P<0.001).Compared with Model group,the serum SCF content of Domperidone group and four acupuncture groups increased in different degrees,but the change rule was different,and only Domperidone group had significant difference compared with Model group(P<0.05).The content of c-kit in Model group was significantly lower than that in Normal group(P<0.01).The content of serum c-kit in PC6 group and BL21 group was significantly higher than that in Model group(P<0.05).The content of c-kit in other groups was higher than that in Model group,but there was no significant difference(P>0.05).5.Results of serum brain-gut peptide: The CCK level in Model group was significantly higher than that in Normal group(P<0.05).The levels of CCK in PC6 group and ST36 group were significantly lower than those in Model group(P<0.05,P<0.01).There was no significant difference between the other groups(P>0.05).The level of serum GAS in Model group was significantly lower than that in Normal group(P<0.05).Compared with Model group,the serum GAS content of other groups increased,but there was no significant difference(P>0.05).The level of Ghrelin in Model group was significantly lower than that in Normal group(P<0.001).The serum Ghrelin content of the other groups was higher than that of Model group,but there was no significant difference(P>0.05).The content of serum MTL in Model group was lower than that in Normal group,while that in the other groups was higher than that in the model group,but there was no significant difference(P>0.05).6.Results of PET/CT(1)Compared with the normal group,the positive activated brain areas in the model group were mainly concentrated in hypothalamus tuberal region,pontine basilar part of pons,midbrain tegmentum of midbrain,septal area,pontine tegmentum of pons,hypothalamus preoptic region,olfactory bulb,striatum,supraoptic region,visual cortex,olfactory tubercle,accumbens nucleus,midbrain substantia nigra,supraoptic region,medulla oblongata,mammillary region,bed nucleus of stria terminalis,cerebral peduncle,orbital cortex,thalamus midline nucleus group,third ventricle,tenia tecta,thalamus medial nucleus group,capsule,amygdaloid body,corpus callosum,dorsal peduncular cortex,anterior commissure,hippocampus,red nucleus,piriform cortex,thalamus lateral nucleus group,dentate gyrus,retrosplenial cortex,infralimbic cortex,olfactory tract,olfactory cortex,anterior nucleus group,blood vessel,middle cerebral artery,subthalamic nucleus.The negative activated areas were mainly concentrated in auditory cortex,sensory cortex,olfactory cortex,temporal association cortex,cerebellum poaterior lobe of cerebellum,parietal cortex posterior area,cerebellum cerebellar nucleus,cerebellum anterior lobe of cerebellum,insular cortex.(2)The Domperidone group had a decreased effect on the brain regions where glucose metabolism was abnormally elevated in the model group,such as thalamus lateral nucleus group,striatum,orbital cortex,olfactory bulb,olfactory tubercle,capsule,piriform cortex,anterior nucleus group,bed nucleus of stria terminalis,anterior commissure.The Domperidone group had an increased effect on the brain regions where glucose metabolism abnormally decreased in the model group,such as cerebellum poaterior lobe of cerebellum,cerebellum cerebellar nucleus,cerebellum anterior lobe of cerebellum.(3)The CV12 group had a decreased effect on the brain regions where glucose metabolism was abnormally elevated in the model group,such as medulla oblongata,corpus callosum,pontine tegmentum of pons,striatum,pontine basilar part of pons,midbrain tegmentum of midbrain,thalamus medial nucleus group,septal area,orbital cortex,hypothalamus tuberal region,amygdaloid body,supraoptic region.The CV12 group had an increased effect on the brain regions where glucose metabolism abnormally decreased in the model group,such as olfactory cortex.(4)The BL21 group had a decreased effect on the brain regions where glucose metabolism was abnormally elevated in the model group,such as corpus callosum,striatum,orbital cortex,third ventricle,hippocampus,dentate gyrus.The BL21 group had an increased effect on the brain regions where glucose metabolism abnormally decreased in the model group,such as olfactory cortex,piriform cortex,temporal association cortex,sensory cortex.(5)The PC6 group had a decreased effect on the brain regions where glucose metabolism was abnormally elevated in the model group,such as orbital cortex,striatum,corpus callosum,third ventricle,hippocampus,septal area.(6)The ST36 group had a decreased effect on the brain regions where glucose metabolism was abnormally elevated in the model group,such as corpus callosum,hippocampus,striatum,medulla oblongata,visual cortex,third ventricle,dentate gyrus,pontine tegmentum of pons,septal area,thalamus medial nucleus group,capsule,amygdaloid body,hypothalamus tuberal region.The ST36 group had an increased effect on the brain regions where glucose metabolism abnormally decreased in the model group,such as olfactory cortex,cerebellum anterior lobe of cerebellum,auditory cortex,temporal association cortex.Conclusion:1.Acupuncture at the acupoints selected according to the part can adjust the hyperglycemia state of diabetic gastroparesis and has a certain hypoglycemic effect.The effect of acupoint selection according to the part of acupuncture on gastric emptying is not the same,in which ST36 and PC6 have obvious promoting effect on gastric emptying.2.Acupoint selection according to the part of acupuncture can improve the pathological morphology of gastric antrum in diabetic gastroparesis rats,but the difference is not obvious.3.Acupoint selection according to the part of acupuncture can improve the ultrastructure of ICCs in gastric antrum of diabetic gastroparesis rats,but the difference is not obvious.4.Acupuncture at acupoints selected according to the part can increase the content of SCF/c-kit signal pathway related protein in diabetic gastroparesis rats,and play a role in promoting gastric movement,but the difference is not obvious.5.Acupuncture at acupoints selected according to the part can adjust the content of brain gut peptide in the serum of diabetic gastroparesis rats,ST36 and PC6 have stronger inhibitory effect on CCK secretion,while CV12 and BL21 have no significant effect.In addition,the secretion of ghrelin was also different between remote acupoint selection and local acupoint selection.6.In the process of glucose metabolism in different brain regions of diabetic gastroparesis rats,it can be observed that there are great differences in the intervention brain regions between different acupoints.In the model group,most of the brain areas with increased glucose metabolism were in the frontal cortex.In the model group,the brain area with increased glucose metabolism decreased mainly in the limbic system.7.There is difference in the effect of acupoint selection according to the part on gastric movement of diabetic gastroparesis rats.The effect of distal acupoint selection on gastric motility was more obvious.The mechanism of the effect difference may be through the different stimulation of brain area,through the brain gut axis pathway to the gastrointestinal tract,regulating gastric movement. |
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