Study On The Mechanism Of Interstitial Cells Of Cajal Participated In In Acute Acalculous Cholecystitis

Objective:Acute calculous cholecystitis(Acute Acalculous Cholecystitis,AAC)was a sudden onset,progress rapidly and higher complications and mortality disease.Gallbladder smooth muscle contraction weakened was one of the main characteristics of AAC.Interstitial Cells of Cajal(Interstitial Cells of Cajal,ICCs)that were distributed in biliary system has produced and spread the gallbladder spontaneous rhythm,regulated the gallbladder movement,caused the gallbladder contraction function.In AAC,quantity,distribution and morphological structure of ICCs were changed,and whether ICCs quantity,distribution,morphological structure can be restored or not when inflammation was reduced or eliminated,it is not clear.This study aims to research quantity,distribution,morphological structure change in ICCs in AAC and inflammation relieved.Methods:250-350g guinea pigs were randomly divided into the control group(the Sham Operated Group,Sham Group),the Common Bile Duct Ligation 24 hours Group(the Common Bile Duct Ligation-24h Group,CBDL-24h Group),the Common Bile Duct Ligation 48 hours Group(the Common Bile Duct Ligation-48 h Group,CBDL-48h Group),the Sham Group was only on laparotomy,the study Groups all were on CBDL.The animals in the study group were subjected to bile duct ligation and then to laparotomy and cholecystectomy at 24 and 48 hours after surgery and measured each animal gallbladder bile capacity,general pathology and HE staining were evaluated degree of AAC inflammation.Secondly,immunohistochemistry and immunofluorescence chemical,TdT-mediated dUTP Nick-End Labeling,transmission electron microscopy(TEM)to observe of gallbladder ICCs distribution and morphological structure changes,and western bolt,real-time fluorescence quantitative polymerase chain reaction(RT-PCR)to observe SCF,c-kit protein and mRNA expression level,SCF/c-kit signal pathway changes.Then,observed the distribution of each part of the gallbladder ICCs groups in animals for clarify the ICCs distribution change in gallbladder inflammation process.Finally,injecting Rabbit anti-polymorphonuclear antibody(Rabbit anti-Rat PMN)advance in AAC disease animal model,then,the Sham Group was only on laparotomy,the CBDL-24h Group,the CBDL-48h Group,anti-PMN Common Bile Duct Ligation-24h Group(anti-PMN CBDL-24h Group)and anti-PMN Common Bile Duct Ligation-48h Group(anti-PMN CBDL-48h Group)all were on CBDL.The animals in the study group were subjected to bile duct ligation and then to laparotomy and cholecystectomy at 24 and 48 hours after surgery.Observed histopathology that were evaluated degree of AAC inflammation,gallbladder tissue inflammation score,gallbladder ICCs distribution and morphological structure changes and SCF,c-kit protein and mRNA expression level,SCF/c-kit signal pathway changes in all groups.Results:After CBDL,with prolonged biliary obstruction,guinea pig gallbladder volume was enlarged gradually,bile was cloudy and floccule was formation in the gallbladder.Gallbladder tissue histopathology showed edema,blood-expanded hyperemia,and inflammatory cell infiltration.Gallbladder tissue inflammation scores is higher(P<0.01).Besides,the number of gallbladder ICCs was decrease and cells apoptosis was increased,cell ultrastructure was damage,SCF/c-kit signaling pathways was down-regulation(all P<0.05).Gallbladder ICCs number was decrease from the neck of gallbladder to the bottom of gallbladder,the quantity of ICCs in each part of the gallbladder in AAC groups were lower than the control group(all P<0.05).After injected anti-polymorphonuclear antibody for animals in advance,pretreatment groups animals gallbladder histopathology,gallbladder tissue inflammation score were all lower than the non-pretreatment groups of animals at the same time(P<0.05).And after gallbladder inflammation relieved,gallbladder damaged ICCs could be repaired in morphology and ultrastructure,the quantity of ICCs were increased,cell apoptosis was decreased,SCF/c-kit signaling pathways was up-regulation,and that may had an effect on the gallbladder contraction dysfunction recovery(all P<0.05).Conclusion:Gallbladder ICCs morphological structure was damage,cell apoptosis were increased,the quantity of ICCs was decreased in AAC,SCF/c-kit signaling pathways was down-regulation,and that may had an effect on cell function,it may be one of the main causes for gallbladder dynamic obstacles occurred in AAC early stage.When gallbladder inflammation relieved,the number of ICCs,and the morphological structure could be restored,suppressed SCF/c-kit signaling pathways was up-regulation,and it’s possible to affect the cell function and gallbladder contraction dysfunction recovery.