Research On The Influence Of Serum Containing Shaoyaoganchao Decoction On [ca²⁺]i In So Cells Of Rabbit With Hypercholesterolemia

Background：The sphincter of oddi is the most important anatomic structure in thebound fraction of cholecyst，pancreas and duodenum，and the only outflow tract ofbile run out of biliary passage.SO is compared to the strobe of bile and pancreatic juice.It consists of four parts,biliary duct sphincter,pancreatic duct sphincter, ampullarysphincter and longitudinal muscles.SO doesn‘t belong to the Small Intestinal SmoothMuscle, on the4th week of Human Embryo, biliary ducts and pancreiatic duct joinedinto ampullar in the form of Mesenchymal Stem before joined with duodenum.on the6th week, Small Intestinal Smooth Muscle formed,on the10th week, MesenchymalStem turned into myoideum.Research finds SO has the characteristic of Forward musclecontractions.The muscle contractions has a synchronization with duodenum,but the twoeach other is irrelevant.With this feature,SO can accommodate the excretion of bile andpancreatic juice，maintain the normal direction flow and the sterile environment ofbiliary tract.So the physiological function of SO has an affinity with Biliary dynamics.Sphincter of Oddi dysfunction is a common clinical disease, according to thePathological Mechanism,SOD can be divided into sphincter of Oddi stenosis andsphincter of Oddi dyskinesia,the essence of the sphincter of Oddi dyskinesia is theContinuously spasm of SO. Professor Wei Jing-guo has confirmedHypercholesterolemia can make SO produces a change on motivity,and can induce SODof Rabbit. As early as in1887, Italian physiologist Oddi found SO, and pointed out thatabdominal pain, fever and jaundice of unknown origin on Some patients maybe causedby SO lesion. After more than20years study at home and abroad, SOD is the Leadinglink and the Key secondary Pathological physiology change of the pathogenesis of mostbiliary diseases.People both at home and abroad do more research on the movement mechanism of vascular smooth muscle and gastrointestinal Smooth Muscle，but they do less researchon the movement mechanism of SO，it has been reported in the literature thatpaeoniflorin can diastole gastrointestinal Smooth Muscle in various ways.The Recentresearch has confirmed that paeoniflorin， which is the chief constituent ofshaoyaoganchao decoction,can diastole vitro SO of Rabbit.The mechanism ofpaeoniflorin diastole gastrointestinal Smooth Muscle is to reduce [Ca²⁺]i，is the samemechanism for shaoyaoganchao decoction to diastole SO，it has not been understandnow.In order to do further research on the mechanism of shaoyaoganchao decoction，weused SO Cells of Rabbit with Hypercholesterolemia loaded with serum containingshaoyaoganchao decoction，determine [Ca²⁺]i in SO cells by means of laser scanningconfocal microscope，so then to Clarify the mechanism of shaoyaoganchao decoctiondiastole SO Cells of Rabbit with Hypercholesterolemia at a cellular level，and providethe basis for clinical treatment of SOD with shaoyaoganchao decoction to some extent.Objective:1.To investigate the influence of high Cholesterol On [Ca²⁺]i in SO Cells ofRabbit with Hypercholesterolemia.2. To investigate the influence of serum containing shaoyaoganchao decoction On[Ca²⁺]i in SO Cells of Rabbit with Hypercholesterolemia.Methods:1.Mature New Zealand white rabbits were randomly divided into2groups: controlgroup, hypereholesterolemia group. control group were fed with standard diet,hypereholesterolemia group were fed with standard diet mixed with cholesterol, Raiseeight weeks to Induce hypercholesterolemia.2.Twenty four mature New Zealand white rabbits were randomly divided into8groups，3rabbits in each groups,they were all administered intragastrically withshaoyaoganchao decoction, twice a day for3days。Blood samples in each groups werecollected respectively from femoral artery after5,15,30,45,60,90,120,180minutes of thelast intragastric administration，use high performance liquid chromatography to measurepaeoniflorin and glycyrrhizic acid contents in blood samples，drawing medicine-timecurve to choose when is the best time to collect blood samples.3. Eight mature New Zealand white rabbits were randomly divided into2groups,control group, experiment group. control group were administered intragastrically withdistilled water, experiment group were administered intragastrically withshaoyaoganchao decoction, twice a day for3days,and75minutes after the last intragastric administration,blood was drown from femoral artery,and separate the serum,diluted to different concentrations and Inactivate complement in56℃constant waterbath for30minutes, sterilized by filtration with0.22μm of filter membrane,then putthem into the usual refrigerator at-20℃.4.Isolate the SO from the Rabbit with Hypercholesterolemia, obtaine SO cells bymeans of enzyme digestion, then primary culture and subculture were used, Identify thecells by Immunohistochemistry.loaded the cells with Fluo-3/AM, investigate [Ca²⁺]I bymeans of confocal laser microscope.Results:1. The hypereholesterolemia group has a higher [Ca²⁺]i in SO Cells of Rabbit thanthe control group;2.[Ca²⁺]i in SO cells of rabbit with hypercholesterolemia load with serumcontaining shaoyaoganchao decoction is lower than in SO cells loaded with normalserum，and has a concentration dependent.Conclusion:1.SO cell is anchorage-dependent cell with fusiform shape;2. Cholesterol can increase [Ca²⁺]i in SO Cells of Rabbit;3.Serum containing shaoyaoganchao decoction can decrease [Ca²⁺]i in SO Cellsof Rabbit with Hypercholesterolemia and has a concentration dependent;4.The change of [Ca²⁺]i in SO Cells can cause the contraction and relaxation ofSO.