| This paper mainly researches the laxatives effects of Atractylodes seven particles granules on STC mice, and studies the changes of colon inhibitory neurotransmitter NO, VIP, excitatory neurotransmitter Ache, SP,ICC, C-kit, 5-HT4, AQP8 and so on in mice with this type of constipation under the intervention of Atractylodes seven particles granules, in order to find the possible mechanism of Atractylodes seven particles granuless in the treatment of STC. It not only provide experimental basis for the treatment of STC in traditional Chinese medicine, but also provides theoretical basis for the treatment method "Supplementing Qi and Nourishing Yin,Promoting qi circulation and transformation" of STC. At the same time, it also provides a theoretical reference for clinical treatment.The first part: preparation of STC modelPurpose:replicate the STC mouse model.Methods: 112 mice were randomly divided into blank group, model 1,2,3,4,5,6 group, a total of 7 groups,16 rats in each group, half male and half female. Model mice subcutaneous injection of morphine hydrochloride 2.5mg.kg-1.d-1, establish STC model, and the blank group was injected with the same amount of normal saline at a total of 45 d. measure dry weight of mice in 24 h before modeling. During the modeling period, the signs of mice were observed. After 45 d, measure the 24 h fecal dry weight of mice. Six mice were randomly selected from each group, half male and half female, and calculate intestinal propulsion rate by cervical dislocation method. In summary, verify if the STC mice model was established successfully.Results: there was no death in 112 mice. Compared with the blank group, the activity of mice in each group was less than that in the model group. The mice in each group had no significant difference in diet, urine and hair color.Before building the model. Compared with the blank group, the body weight of model mice in each group has no significant difference(P> 0.05), no statistical significance. 45 days after modeling, compared with blank group, the model group mice had no obvious weight loss, no significant difference(P> 0.05), no statistical significance. Before building the model. Compared with the blank group, the body weight of model mice in each group no significant difference(P> 0.05), no statistically significant. After modeling, compared with the blank group, model group mice stool was dry, a separation of hard agglomerate agglomerate or feces, significantly reduced the number of defecate, defecation weight was significantly reduced, there was significant difference(P< 0.05), and statistically significant. Compared with the blank group, the model group 1,3,4,5,6 mice intestinal propulsion rate was significantly reduced(P< 0.01), there was a high degree of statistical significance; model group 2 mice intestinal propulsion rate was significantly reduced(P< 0.05), and there was statistical significance.Conclusion: according to the dose of 2.5mg.kg-1.d-1, making mold using the method of Subcutaneous inject Morphine Hydrochloride Injection in 45 consecutive days, can successfully replicate the STC model, and this method is easy to operate and has high security. The model mice showed obvious dry stool, lump or mass like the separation of feces, feces number decreased significantly, which was meeting standard in change of character of stool. Model group mice intestinal transmission function was significantly lower, meets the STC clinical diagnosis standard. To sum up, the STC model was successfully copied.The second part: research on the effect of Atractylodes seven particles granules on motility in STC model micePurpose: study the effect of the STC model in mice under the prevention ofAtractylodes seven particles granules.Methods: After the STC mouse model successfully established, the remaining 70 mice, and the blank group stays unvaried. The rest 60 mice were randomly divided into model group, low, middle and high dose group of Atractylodes seven particles granules, Maren soft capsule group, Mo Sapele will group, 10 rats in each group, half male and half female. In the blank group, model group was given normal saline to fill the stomach, and the remaining groups were given corresponding drugs to fill the stomach in 14 d. Observe the influence of Atractylodes seven particles granules on the first defecation time and the intestinal propulsion rate of STC mice model.Results: there was no death in 70 mice;Compared with the blank group, there was no significant difference in the activities, diet, urine and hair color of mice in each group.Compared with the model group, after administration, the mice in each group of mice were less dynamic symptoms of fatigue symptoms gradually improved, fecal water increased, the number of bowel movements increased, sausage shaped or clumps like smooth and soft. Before administration, compared with the blank group, the model group had no significant difference in body weight(P> 0.05), and had no significant difference.Compared with the model group, Atractylodes seven particles granules were lower, middle and high dose group, Maren soft capsule group, Mo Sapele weight group no significant difference(P> 0.05), no statistical significance.After 14 days administration, compared with the blank group, the model group had no significant difference in body weight(P> 0.05), and had no significant difference. Compared with the model group, Atractylodes seven particles granules were lower, middle and high dose group, Maren soft capsule group, Mo Sapele weight group no significant difference(P> 0.05), no statistical significance.Compared with the blank group, the model group was significantly longer(P< 0.01), which was the first time in the model group, which was highly significant.Compared with the model group, Atractylodes seven particles granules were lower, middle and high dose group, Maren soft capsule group, Mo Sapele group were the first black stool time significantly shortened(P< 0.01), was highly statistically significant.Compared with the seven Atractylodes compound granules of high dose group, seven Atractylodes compound granules of low, middle dose group, Maren soft capsule group, Mo Sapele were ranked first black stool time had no significant difference(P> 0.05). No statistical significance. Compared with the blank group, the model group significantly reduced the intestinal propulsion rate(P< 0.01), which was highly significant; Compared with the model group, Atractylodes seven particles granules were lower, middle and high dose group, Maren soft capsule group, Mo Sapele group mice intestinal propulsion rate increased significantly(P< 0.01), was highly statistically significant. Compared with the seven Atractylodes compound granules of high dose group, Atractylodes seven particles granules of low, middle dose group, Maren soft capsule group, Mo Sapele group mice intestinal propulsive rate of no significant difference(P> 0.05). No statistical significance.Conclusion: 1 Atractylodes seven particles granules can improve the STC model mice.2.Atractylodes seven particles granules can achieve the treatment of STC model mice by promoting defecate function. 3. Atractylodes seven particles granules can achieve the effect of treating STC model mice through the small intestine to accelerate the speed, improve small intestinal propulsion rate and promote intestinal peristalsis of STC mice. 4.Atractylodes seven particles granules can promote STC mice defecation, defecation function, promote intestinal peristalsis and Maren soft capsule and Mo Sapele.The third part: the research on the multi intervention mechanism of Atractylodes seven particles granules on the STC model micePurpose: study the colon inhibitory neurotransmitter NO, VIP, excitatory neurotransmitter Ache, SP,, ICC and c-kit,, 5-HT4, AQP8, and other changes in the STC model mice under the intervention of Atractylodes seven particles granules, in order to find the possible mechanism of Atractylodes seven granules in the treatment of STC.Method: Detect the expression of NO, VIP, Ache, SP, ICC and AQP8,, 5-HT4, c-kit, excitatory neurotransmitter in mice colon tissue of mice with STC model using the ELISA, RT-PCR, HE staining and immunohistochemical methods when using the Atractylodes seven particles granulesResults: ELISA test results analysis: compared with the blank group, the model group content was significantly increased(P< 0.05), there was statistical significance;The VIP content of the model group increased significantly(P< 0.01), a highly statistically significant; the ache content of the model group decreased significantly(P< 0.01), a highly statistically significant; the content of SP in model group decreased significantly(P< 0.01), a highly statistically significant. Compared with the model group, Atractylodes seven particles granule low dose group, content has no significant difference(P> 0.05), no statistically significant; Atractylodes seven particles granules dose group, no content decreased significantly(P< 0.05) and was statistically significant.Atractylodes seven granules of high dose group, Maren soft capsule group, Mo Sapele group, the no content decreased significantly(P< 0.01), a highly statistically significant. Seven Atractylodes compound granules of low, mid and high dose groups, Maren soft capsule group, Mo Sapele group, the content of VIP decreased significantly(P< 0.01), a highly statistically significant. Atractylodes seven granules of low dose of ache content no significant difference(P> 0.05). No statistical significance; Atractylodes seven particles granules in high dose group, Maren soft capsule group, Mo Sapele group ache content increased significantly(P< 0.01), highly statistically significant. Atractylodes seven particle agent of low, mid and high dose groups, Maren soft capsule group, Mo Sapele group SP content increased significantly(P< 0.01), highly statistically significant. Compared with the high dose of Atractylodes seven particles granules group, the content of middle dose of Atractylodes seven particles granules group was significantly increased(P< 0.01), highly statistically significant; Maren soft capsule group, Mo Sapele group, the content decreased significantly(P< 0.01), and has a highly statistically significant. Seven Atractylodes compound granules of low, VIP content in middle dose group increased significantly(P< 0.01), highly statistically significant. Ma Renruan capsule group, Mo Sapele Group VIP content no significant difference(P> 0.05), no statistically significant. Seven Atractylodes compound granules of low, middle dose group, Maren soft capsule group ache content have no significant difference Different(P> 0.05), no statistically significant; Mo Papele ache content in Sapele group increased significantly(P< 0.05) and was statistically significant. Seven Atractylodes compound granules of low, middle dose group, Maren soft capsule group, Mo Sapele group SP content decreased significantly(P< 0.01), a highly statistically significant.The analysis of RT-PCR results: compared with the control group, the content of c-kit in model group decreased significantly(P< 0.01), highly statistically significant. 5-HT4 of model group was significantly decreased(P< 0.01), a highly statistically significant. AQP8 in model group increased significantly(P< 0.01), a highly statistically significant. Compared with the model group, low, middle dose of the Atractylodes seven particles granules group, Maren soft capsule group c-kit content no significant difference(P>0.05), no statistically significant; Atractylodes seven particles granules high-dose group c-kit content increased significantly(P< 0.01), a highly statistically significant; Mo Sapele group and c-kit content increased significantly(P< 0.05), was statistically significant. low dose of Atractylodes seven particles granules, Maren soft capsule group 5-HT4 content no significant difference(P> 0.05), no statistically significant; Atractylodes seven particles granules in high dose group, Mo Sapele group 5-HT4 content significantly Elevated(P< 0.01), highly statistically significant; Atractylodes seven granules of low dose of AQP8 content no significant difference(P> 0.05), no statistical significance; Atractylodes seven particles granules in high dose group, Maren soft capsule group, Mo Sapele group AQP8 content decreased significantly(P< 0.01), and has highly statistically significant. Compared with the high dose of Atractylodes seven particles granules group, the low-dose of seven Rhizoma Atractylodis Macrocephalae granules group c-kit content decreased significantly(P< 0.01), highly statistically significant; Atractylodes seven particles granules dose group c-kit content decreased significantly(P< 0.05), was statistically significant; Maren soft capsule group, Mo Sapele group c-kit content no significant difference(P> 0.05), no statistically significant. Seven Atractylodes compound granules of low, middle dose group, Maren soft capsule group and Mo Sapele group 5-HT4 content no significant difference(P> 0.05), no statistically significant. Atractylodes seven particles granules low-dose AQ groups P8 increased significantly(P< 0.01), a highly statistically significant; Atractylodes seven particles granules in the dose group, Maren soft capsul group, Mo Sapele AQP8 in Sapele group no significant difference(P> 0.05). No statistical significance.The analysis of HE staining: model group mice colon tissue nerve cells decreased, and can also see infiltration of inflammatory cells in the mucous layer, score for + + or +++. mice in each treatment group colonic neurons was restored and colon tissue infiltration of inflammatory cells reduced, scoring of ± or-.The analysis of immunohistochemistry: compared with the blank group, model group, no average optical density increased significantly(P< 0.05), was statistically significant. The average optical density of VIP in model group increased significantly(P< 0.01), a highly statistically significant. The average optical density of ache in model group decreased significantly(P< 0.01), a highly statistically significant. The average optical density of SP in model group decreased significantly(P< 0.01), a highly statistically significant. The average optical density of c-kit in model group decreased significantly(P< 0.01), a highly statistically significant. The average optical density of model group 5-HT4 significantly reduced(P< 0. 01), there was a high degree of statistical significance. The average light density of the model group was significantly increased(P< 0.01), and there was a highly significant statistical significance. Compared with the model group, low, middle dose of Atractylodes seven particles granules group, no average optical density has significant difference(P> 0.05). No statistical significance; Atractylodes seven granules of high dose group, Maren soft capsule group, Mo Sapele were no mean optical density decreased significantly(P< 0.01), there is a high degree of statistical significance. Atractylodes seven granules of low dose group of VIP average optical density of no significant difference(P> 0.05). No statistical significance; Atractylodes seven particles granules in high dose group, Mo average optical density of Sapele VIP group decreased significantly(P< 0.01), a highly statistically significant. Ma Ren soft capsule Group VIP flat Are optical density decreased significantly(P< 0.05), was statistically significant. Atractylodes seven particles granules low-dose ache light density without significant difference(P> 0.05), no statistically significant; Atractylodes seven particles granules in the dose group, Ma Renruan capsule group ache average optical density increased significantly(P< 0.05), was statistically significant. Atractylodes seven granules of high dose group, Mo average optical density of ache in Sapele group increased significantly(P< 0.01), highly statistically significant. Atractylodes seven granules of low dose SP average optical density of no significant difference(P> 0.05). No statistical significance; Atractylodes seven Granules in high dose group, Maren soft capsule group, Mo Sapele average optical density of SP increased significantly(P< 0.01), highly statistically significant.Atractylodes seven particles granules in high dose group, Maren soft capsule group, Mo Sapele average optical density of SP increased significantly(P< 0.01), a highly statistically significant. Seven Atractylodes compound granules of low, middle dose group, Maren soft capsule group c-kit average optical density of no significant difference(P> 0.05). No statistical significance; the average optical density of Atractylodes seven granules of high dose group of c-kit was significantly increased(P< 0.01), a highly statistically significant; Mo Sapele group c-kit average optical density increased significantly(P< 0.05), was statistically significant. Atractylodes seven granules of low dose 5-HT4 flat Were light density had no significant difference(P> 0.05), no statistically significant; Atractylodes seven particles granules in high dose group, Maren soft capsule group 5-HT4 average optical density increased significantly(P< 0.05), was statistically significant; Mo Sapele group 5-HT4 average optical density increased significantly(P< 0.01), highly statistically significant; Atractylodes seven granules of low dose group of AQP8 average optical density of no significant difference(P> 0.05). No statistical significance; Atractylodes seven particles granules in high dose group, Maren soft capsule group, Mo Sapele group AQP8 average optical density significantly increased(P< 0.05), there is statistical significance. Compared with high dose of Atractylodes seven particles granules of group low, middle dose of Atractylodes seven particles granules of group, Maren soft capsule group, Mo Sapele average optical density has no significant difference(P> 0.05), no statistically significant. Atractylodes seven granules of low dose group of VIP average optical density increased significantly(P< 0.05), was statistically significant. Atractylodes seven particles granules in the dose group, Maren soft capsule group, Mo Sapele Group VIP average optical density of no significant difference(P> 0.05). No statistical significance. Atractylodes seven granules of low dose group of ache average optical density decreased significantly(P< 0.05), statistical significance; Atractylodes seven particles granules in the dose group, Maren soft capsule group, Mo Sapele group ache average optical density of no significant difference(P> 0.05). No statistical significance. Atractylodes seven granules of low dose SP group average optical density decreased significantly(P< 0.05), was statistically significant; Atractylodes seven particles granules in the dose group, Maren soft capsule group, Mo Sapele group SP average optical density of no significant difference(P> 0.05). No statistical significance. Seven Atractylodes compound granules of low, middle dose group, Maren soft capsule group, Mo Sapele group c-kit average optical density without significant difference(P> 0.05). No statistical significance Righteousness. Seven Atractylodes compound granules of low, middle dose group, Maren soft capsule group, Mo Sapele group 5-HT4 no significant difference(P> 0.05), no statistically significant. Atractylodes seven granules of low dose group of AQP8 average optical density increased significantly(P< 0.05), was statistically significant. Atractylodes seven particles granule medium dose group, Maren soft capsule group, Mo Sapele group AQP8 average optical density of no significant difference(P> 0.05). No statistical significance.Conclusion: 1. Atractylodes seven particles can achieve the effect of treating STC model mice through reducing the content of NO and VIP to strengthen gastrointestinal smooth muscle contraction and the gastrointestinal peristalsis. 2.Atractylodes seven particles can achieve the effect of treating STC by increasing the content of Ache and SP,exciting smooth muscle and stimulating intestinal motility. 3.Atractylodes seven particles can achieve the effect of treating STC by increasing the c-kit content of ICC, and speeding up the transfer of electrical excitation, enhancing colonic motility. 4.Atractylodes seven particles can treat STC by increasing the 5-HT4 content, to promote intestinal peristalsis. 5.Atractylodes seven particles can cure STC by decreasing the content of AQP8, to reduce the intestinal water absorption. 6. The high dose of Atractylodes seven particles has the most significant therapeutic effect of STC. |
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