| 4Background: Gallbladder as part of the human organs,and is mainly used for concentrated stores bile excrete bile,it is also the important parts of the biliar.The biliary disease also occupies certain proportion in the disease.Chronic calculous cholecystitis belongs to biliary dynamics function abate Chronic disease.As Hinescu ME [1]for the first time found Cajal interstitial cells(ICC)in biliary tract,also for the future study of biliary dynamics mechanism provides a direction.The discovery of the ICC in the gastrointestinal tract and its research progress of mechanisms is more deeply than on biliary dynamics,it also provides a research basis for study of biliary dynamic mechanism of and reference[2].According to the research shows that the ICC is gastrointestinal pacemaker cells,adjust the rhythm of gastrointestinal smooth muscle,also is the basis of the normal gastrointestinal movement condition.Gap junction protein 43,45(Connexin43,45,Cx43,Cx45)constitute the intercellular gap junction(GJ)channel,the basic structure and function of a large family of membrane proteins are relatively widely distributed in two kinds of protein[3].And GJ is between adjacent cells of small molecules and special electrochemical signal transmission channel,has moderate synergistic reaction of adjacent cells.With the progress of the ICC-SMC network research,existing related studies have shown that Cx43,Cx45 exists in gastrointestinal Cajal interstitial cells(ICC)and smooth muscle cells(SMC)[1],participate in the regulation of gastric bowel movement.And the gallbladder wall layer structure and lining structure of the gastrointestinal tract are similar,Gallbladder whether there are Cx45,Cx43 expression,whether the relationship between ICC,Cx45 and Cx43 in the gastrointestinal tract is similar,to participate in the regulation of biliary dynamics is unknown.From this point of view,we study the relationship between Cx43,Cx45 and ICC in gallbladder motility and its correlation.Artemisia capillaris soup as a traditional Chinese medicine(TCM),mainly used for heat,moisture,and the treatment of liver,gallbladder and other conditions[4?5],our study found[6] that in the treatment of chronic calculous cholecystitis has obvious effect,to deepen the understanding between the drug and chronic calculous cholecystitis and gall bladder the ICC,the relationship between the need to further research.Objective:Section 1: Through the case analysis,artemisia capillaris decoction in the treatment of chronic calculous cholecystitis patients of gallbladder contraction function changes.Section 2: This experiment through the given experimental guinea pigs with high cholesterol feed stone method [21],chronic calculous cholecystitis chronic model was established,and in 8 weeks after treated group Yinchenhao Decoction orally.The gallbladder was stained by HE,immunohistochemistry and immunofluorescence.The changes of the number and shape of gallbladder ICC were observed.The expression changes of Cx43,Cx45 protein and ICC were observed.To investigate the interaction between Cx43,Cx45 and ICC,and the effects of Yinchenhao Decoction on Cx43,Cx45 and ICC in the gallbladder.Methods:Section 1:1.Selected in the First Affiliated Hospital of Dalian Medical University,Dalian Medical University Second Affiliated Hospital of nearly two years of chronic calculous cholecystitis,in accordance with the diagnostic criteria for the disease index screening,were randomly divided into Artemisia Capillaris Soup in the treatment group and Heat cholagogue particles in the treatment group,and select no correlation of biliary system diseases volunteers as control group and study together.2.Relevant indicators by ultrasound measurement,and finally to obtain data,statistical analysis,draw the conclusion.Section 2:1.Select experiment with common guinea pigs,are all male,3-4 months old,weight 250-300 grams,after randomization,divided into three group and normal group,model group and treatment group.After the breeding and treatment.2.Line respectively in each group the gallbladder,frozen section,after fixation,dehydration,dyeing steps HE staining,and under the light microscopy to observe the pathology of the gallbladder constituting the change,after in the department of pathology,the first principle of independent double-blind,please the teacher evaluation,validation model was successful.3.Groups of gallbladder tissue,were taken by fixation,embedding,incubation and so on after immunohistochemical steps and immunofluorescence double standard immune steps,seal save,observation group the expression of Cx43 and Cx45 in gallbladder muscle layer and the relationship between the ICC collect images useful and statistical analysis.Results:Section 1:1.Artemisia capillaris soup and Heat cholagogue particles in the two groups before treatment respectively and the control group compared to the gallbladder contraction rate decreased significantly(P < 0.05),the gallbladder contractile function and performance is a bad one.2.Artemisia capillaris soup and Heat cholagogue particles in the treatment of two groups of drugs and treatment before gallbladder contraction rate increased significantly(P < 0.05),indicate that the two drugs have significantly improve the contraction function of gallbladder.3.Artemisia capillaris soup and Heat cholagogue particles after treatment in two groups of drugs compared two groups no significant difference(P > 0.05),but Artemisia capillaris soup compared obvious.Section 2:1.Groups of HE staining showed different structures,based on the change of cell morphology and organization structure,model group in epithelial cells arranged in disorder,epithelial shedding and defect,submucosal edema of the muscle layer of compensatory hypertrophy,a large number of lymphocyte invasion is visible.2.Immunohistochemical results show that in the gallbladder with the expression of Cx43 and Cx45 in the gallbladder muscle was brown and yellow.The expression quantity and color of Cx43 and Cx45 in model group were significantly lower than that in normal group(P<0.05).The expression quantity and color of Cx43 and Cx45 in treatment group were significantly higher than that in model group(P<0.05).3.Immunofluorescence,compared with the normal group,the expression of Cx43,Cx45 and ICC in model group was significantly decreased(P<0.05).Compared with the model group,the fluorescence intensity of Cx43,Cx45 and ICC expression was significantly increased(P<0.05).Conclusion:1.The expression of Cx43,Cx45 protein and Cx45 and Cx43 in the gallbladder muscle tissue was significantly decreased in the patients with chronic cholecystitis.Artemisia capillaris soup after treatment increased significantly2.In chronic calculous cholecystitis formation and structure of the ICC damage and expression decreased significantly,Cx43,Cx45 expression also decreased significantly,Cx43,Cx45 and ICC has a same expression rule,and with biliary dynamics occur corresponding links.3.After the treatment group,the Artemisia capillaris soup has the obvious effect on the treatment of chronic calculous cholecystitis. |
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