| Objective:To investigate the effect of Guirenrunchang Particles on colon neurotransmitters NO,Ache,VIP and SP in STC mice model.To investigate the effect of Guirenrunchang Particles on colon AQP3 in STC mice model.To investigate the effect of Guirenrunchang Particles on the pathological changes of colon tissue in STC mice model.Methods:1 Establishing STC mice model by morphine method.2 Testing the effects of Guirenrunchang Particles on neurotransmitters NO,Ache,VIP and SP by Elisa method.3 Observing the effect Guirenrunchang Particles on the pathological changes of mice colon tissue by HE stain4 Observing the effect Guirenrunchang Particles on AQP3 expression in mice colon tissues by immunohistochemistry.Results:1 Replication of STC mice modelAfter 45 days of molding by morphine method,no mice died in two group,It shows that morphine method is safe and reliable.Compared with Normal Control group,mice in Model Control group were tired and less activities,their back hair was less shiny and frizzled.There was no significant difference between Normal Control group and Model Control group in diet and urine.There was no difference in body weight between Model Control group and Normal Control group.2 Effect of Guirenrunchang Particles on intestinal motility in STC mice model.In the course of administration,72 mice did not die,indicating that the experimental method was safe and reliable.Compared with Normal Control group,mice in Model Control group were tired and less activities,their back hair was less shiny and frizzled.The number of feces particles was less,showing a smaller granular shape and a dry surface.Comparedwith Model Control group,the mice in Low Dosage group,Medium Dosage group,High Dosage group and Positive Control group(mosapride group)showed tired and less activities improved,less shiny and frizzled back hair improved,fecal particle number increased,feces are more watery and the surface is wetter,feces form is smooth and soft.In Low Dosage group,Medium Dosage group,High Dosage group and Positive Control group,Low Dosage group had more granular feces.There was no significant difference in feces between the other three groups,feces surface is wet,shape is shape.There was no significant difference in diet water and urine among Normal Control group,Model Control group,Low Dosage group,Medium Dosage group,High Dosage group and Positive Control group.Weight change before and after administration: Before administration,there was no significant difference in body weight between Normal Control group and Model Control group,Low Dosage group,Medium Dosage group,High Dosage group and Positive Control group(P>0.05).There were no significant differences among Model Control group,Low Dosage group,Medium Dosage group,High Dosage group and Positive Control group(P>0.05).After administration,there was no significant difference in body weight between Normal Control group and Model Control group,Low Dosage group,Medium Dosage group,High Dosage group and Positive Control group(P>0.05).There were no significant differences among Model Control group,Low Dosage group,Medium Dosage group,High Dosage group and Positive Control group(P>0.05).Fecal weight change before and after administration: before administration,compared with Normal Control group,the average dry weight of 24 h feces was significantly reduced in Model Control group,Low Dosage group,Medium Dosage group,High Dosage group and Positive Control group(P<0.01).After 14 days of administration,compared with Normal Control group,the average dry weight of 24 h feces was still significantly reduced in Model Control group,Low Dosage group and Medium Dosage group(P<0.01);The average dry weight of 24 h feces was reduced in High Dosage group and Positive Control group for24h(P<0.05).Compared with Model Control group,the average dry weight of 24 h feces in Medium Dosage group increased(P<0.05).The average dry weight of 24 h feces in HighDosage group and Positive Control group increased significantly(P<0.01).Mice intestinal propulsion experiment: the intestinal propulsive rate of Model Control group was significantly lower than that of Normal Control group(P<0.01);Compared with Model Control group,there was no significant difference in Low Dosage group(P>0.05),the intestinal propulsion rate increased in Medium Dosage group(P<0.05),and significantly increased in High Dosage group and Positive Control group(P<0.01);Compared with High Dosage group,the propulsion rate of Low Dosage group was significantly different(P<0.01),the propulsion rate of Medium Dosage group was significantly different(P<0.05),and the propulsion rate of Positive Control group was similar(P>0.05).3 Effects of Guirenrunchang Particles on multiple mechanisms in STC mice model.Effects of Guirenrunchang Particles on neurotransmitter NO: compared with Normal Control group,the concentration of NO was significantly increased in Model Control group(P<0.01).Compared with Model Control group,NO of blood in Medium Dosage group,High Dosage group and Positive Control group all decreased(P<0.05).Among them,blood NO of High Dosage group and Positive Control group decreased significantly(P<0.01).Effects of Guirenrunchang Particles on neurotransmitter VIP: compared with Normal Control group,the concentration of VIP was significantly increased in Model Control group(P<0.01).Compared with Model Control group,the blood VIP in Medium Dosage group,High Dosage group and Positive Control group all decreased(P<0.05),among which the blood VIP in High Dosage group and Positive Control group decreased significantly(P<0.01).Effects of Guirenrunchang Particles on neurotransmitter SP:compared with Normal Control group,the concentration of SP was increased in Model Control group(P<0.05).Compared with Model Control group,the blood SP of Medium Dosage group,High Dosage group and Positive Control group were significantly increased(P<0.01).Effects of Guirenrunchang Particles on neurotransmitter Ach: compared with Normal Control group,the concentration of VIP was significantly increased in Model Control group(P<0.01).Compared with Model Control group,the blood Ach in Low Dosage group,Medium Dosage group,High Dosage group and Positive Control group were all increased(P<0.05).Among them,the blood Ach of Medium Dosage group,High Dosage group and Positive Control group was significantly increased(P<0.01).Effects of Guirenrunchang Particles on HE staining of mice colon tissues: Goblet cells in normal colon tissues were evenly distributed,without obvious goblet cell cytoplasmic expansion or obvious infiltration of inflammatory cells in the interstitium.In Model Control group,the cytoplasm of goblet cells in the colon tissue was significantly dilated,the cytoplasm was enlarged,and the interstitium was infiltrated by more chronic inflammatory cells dominated by lymphocytes.The distribution of goblet cells in colon tissues of moxabilli group and moxabilli group was relatively uniform,the degree of goblet cell cytoplasm dilation was reduced,a small amount of goblet cell cytoplasm locally increased,and the infiltration of chronic inflammatory cells dominated by lymphocytes was rare in the interstitium.In High Dosage group of Guirenrunchang Particles,the distribution of goblet cells in colon tissues was relatively uniform,the degree of goblet cell cytoplasm dilation was reduced,a small amount of goblet cell cytoplasm was increased locally,and the infiltration of chronic inflammatory cells dominated by lymphocytes was rare in the interstitium,and the effect was better than that in moxabilli group.In Medium Dosage group of Guirenrunchang Particles,the distribution of goblet cells in colon tissues was relatively uniform,the degree of goblet cell cytoplasm dilation was reduced,the local goblet cell cytoplasm was increased,and the infiltration of chronic inflammatory cells dominated by lymphocytes was rare in the interstitium.In Low Dosage group of Guirenrunchang Particles,the distribution of goblet cells in colon tissues was relatively uniform,the degree of goblet cell cytoplasmic dilatation was reduced,the local goblet cell cytoplasmic dilatation was larger and more frequent,and the infiltration of chronic inflammatory cells dominated by lymphocytes was more frequent in the interstitium,and the effect was worse than that in moxabelic group.Effect of Guirenrunchang Particles on AQP3 expression by immunohistochemical detection: the positive AQP3 expression is brownish yellow or dark brown,scattered in thecolonic gland stroma.The positive AQP3 cells in the colon tissues of Normal Control group were brown-yellow,and the positive cells were scattered and distributed in the glandular stroma,with less positive cells and low positive expression.The positive AQP3 cells in Model Control group were brownish yellow or dark brown,and the positive cells were scattered and distributed in the glandular stroma,with more positive cells and high positive expression.The positive AQP3 cells in Positive Control group were brown-yellow,and the positive cells were scattered and distributed in the glandular stroma.The number of positive cells was significantly reduced,and the positive expression was low.The positive AQP3 cells in High Dosage group of Guirenrunchang Particles were brown-yellow,and the positive cells were scattered and distributed in the glandular stroma,and the number of positive cells was significantly reduced,and low positive expression.Compared with Positive Control group,the number of positive cells was significantly lower and the positive degree was decreased.Compared with Positive Control group,the number of positive cells was significantly lower and the positive degree was decreased.The positive AQP3 cells in Medium Dosage group of Guirenrunchang Particles were brown-yellow,and the positive cells were scattered and distributed in the glandular stroma,and the number of positive cells was significantly reduced,and low positive expression.Compared with Positive Control group,the number of positive cells was similar and the degree of positive cells was decreased.In Low Dosage group of Guirenrunchang Particles,the positive AQP3 cells were brown-yellow,and the positive cells were scattered and distributed in the glandular stroma.More positive cells scattered and distributed,a small part of cells were clustered,and the positive expression was high.Effect of Guirenrunchang Particles on AQP3 expression by western blot detection:Compared with Normal Control group,the relative gray value of the Model Control group was significantly increased(P<0.01),indicating that the content of AQP3 in Model Control group was significantly increased.Compared with Model Control group,the relative gray value of AQP3 in the Medium Dosage group,High Dosage group and the Positive Control group all decreased(P<0.05),and the relative gray value of AQP3 in the High Dosage groupand Positive Control group all decreased significantly(P<0.01).Conclusion:1 Establishing STC mice model by morphine method,Model Control group mice were tired and less activities,their back hair was less shiny and frizzled,fecal weight is lighter than Normal Control group,the shape of feces was dry and hard granular,and the number of feces particles was reduced,which was consistent with the state and the change of fecal shape of STC mice model.Mice intestinal propulsion experiment showed that the intestinal transport function of mice in Model Control group was lower than that in Normal Control group,which was consistent with the clinical characteristics of STC,suggesting that the preparation of STC mice by morphine method was successful.STC mice model by morphine method is simple,stable,safe,economical and effective.2 Guirenrunchang Particles can improve fecal properties of STC mice.Compared with Model Control group,Medium Dosage group,High Dosage group and Positive Control group were significantly improved,the mice showed tired and less activities improved,less shiny and frizzled back hair improved,fecal particle number increased,feces are more watery and the surface is wetter,feces form is smooth and soft,the feces of mice in Low Dosage group were also improved,but not as much as those in the other three groups.Guirenrunchang Particles can increase the weight of dry feces of STC mice to reach the effect of treating STC.Medium dosage of Guirenrunchang Particles and high dosage of Guirenrunchang Particles have effects.High dosage of Guirenrunchang Particles is more obvious,which is equivalent to mosapride.Guirenrunchang Particles can accelerate the intestinal propulsion rate of STC mice,which increases at low,medium and high dosage.The improvement of Guirenrunchang Particles at high dosage is the most obvious,which is equivalent to mosapride,indicating that Guirenrunchang Particles can improve the intestinal peristalsis of mice and have the effect of treating STC.3 Guirenrunchang Particles can reduce the content of NO and VIP,increase the content of Ach and SP,stimulate the smooth muscle from the neurotransmitter,and strengthen thegastrointestinal propulsion speed to treat STC.By reducing the expression of AQP3 and the absorption of water by AQP3 in the intestinal tract,Guirenrunchang Particles can achieve the effect of treating STC.The expression of AQP3 in the colon of mice was decreased by medium dosage of Guirenrunchang Particles,high dosage of Guirenrunchang Particles and mosapride In each group,the effect of high dosage of Guirenrunchang Particles on STC was the most significant.In terms of its effect on neurotransmitter,high dosage of Guirenrunchang Particles similar to mosapride,and its effect on AQP3 expression is better than mosapride.The effect of medium dosage and low dosage of Guirenrunchang Particles on STC is weaker than that of mosapride,and the effect of low dosage of Guirenrunchang Particles on STC is not obvious.4 The changes of NO,VIP,Ach and SP by Guirenrunchang Particles were obvious.Combined with the change of intestinal propulsion rate in mice,it is possible that Guirenrunchang Particles could treat STC by affecting NO,VIP,Ach,SP and other neurotransmitters to accelerate intestinal transport.Guirenrunchang Particles had significant effect on the expression of AQP3.The morphology and moistening degree of mice feces were also significantly changed.It is possible that Guirenrunchang Particles could treat STC by inhibiting the expression of AQP3 to reduce the reabsorption of water in the intestinal tract,increase the water content of feces,and improve the peristaltic movement of the intestine. |
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