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Experiment Study on the Effect of Specific COX-2 Inhibitor Celecoxib on Nucleating Proteins-immunoglobulins and Mucin

Author: CaiChenXiao
Tutor: CaiJianTing
School: Zhejiang University
Course: Internal Medicine
Keywords: Cholesterol stones Cyclooxygenase-2 Immunoglobulin Mucin
CLC: R575.6
Type: Master's thesis
Year: 2004
Downloads: 40
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Abstract

Purpose and background of gallstones is a common worldwide, with high disease prevalence rate of 10%. With the improvement of people's living standards, the incidence of cholelithiasis has gradually increased, especially cholesterol gallstone patients is increasing, people are concerned about how the prevention and treatment of cholelithiasis. Cholesterol gallstone formation process by a variety of factors, in addition to closely related to the class of abnormal lipid metabolism and gallbladder factors, its formation mechanism may also include a chronic inflammatory process. That cholesterol gallstone formation must meet the following conditions: liver metabolism, secretion of supersaturated hepatic bile, leading to nuclear factor and suppression of nuclear factor balance between destruction and gallbladder motility, have sufficient time to the formation of stones in the gallbladder bile. Immunoglobulin IgA, IgG, IgM as bile contributed to nuclear factor have been numerous reports in the literature, the immunoglobulin in the blood circulation is the main source of the bile immunoglobulin. The study found that the IgA, IgG, IgM were having a nucleating activity, in particular, is IgG, not only so that the nucleation time is shortened, but also the number and speed of the generated crystals were increased. Mucin is the main component of gallbladder epithelial cells secrete mucus as a contributing nuclear factor has been more scholars confirmed, but the mechanism is not yet fully understood. Mucin core peptides of mucin (MUC) gene encoding a specific MUC expression in patients with cholesterol gallstones gallbladder epithelial cells. Cyclooxygenase (cyclooxygenase, COX) is the rate-limiting enzyme catalyzed metabolism of arachidonic acid to prostaglandin (PG _S). Recent study found that COX There are two subtypes: structure type of COX-1 and inducible COX-2. COX-1 is stably expressed in the gastrointestinal mucosa, platelets, regulate prostaglandin and thromboxane A ₂ (Thronboxane the A ₂, the TXA ₂) formation of a protective mucous membranes, regulate blood vessels to dilate, promoting the role of platelet aggregation; COX-2 with little or almost non-existent in the gastrointestinal mucosa, it may be bacterial lipopolysaccharide (LPS, LPS), interleukin -1 (Interleukin IL-1) and other inflammatory cytokines Fenjiang the days of school dental schools' a master's degree in theory husband induced a sharp increase in the level of the sites of inflammation, and its expression levels associated with the severity of the inflammation, and the inflammatory response are closely relationship. Copy Cat chronic cholecystitis model, Nilsson and other people find that high expression of COX-2 in inflammation of the gallbladder exclusion membrane. A number of experiments show that COX-2 inhibitors can produce a role similar to the multi-body anti-inflammatory drugs (nonsteriodalanti infl ^{a tory drugs, NSAIDs) inhibit prostaglandin synthesis, but its role in the NSAIDs the position only produce anti-inflammatory and analgesic effect, reducing the number of adverse reactions. NSAIDs can reduce the formation of cholesterol gallstones, but while inhibition of COX-2, also inhibits COX-1 activity, reducing the COX-1 of PG synthesis induced by undesirable side reaction caused by the gastrointestinal tract. Selective COX-2 inhibitors reduce these adverse side effects, and has great application prospects. The study found that COX-one involved in the reaction of cholecystitis, selective COX-2 inhibitors can prevent LPS and other inflammatory factors to stimulate the gallbladder cell PG synthesis, and effective treatment of gallbladder disease. Experimental study on whether the selective COX-2 inhibitors in cholesterol gallstone formation process by affecting the inflammatory response to inhibit or reduce calculus formation and mechanism of action, has not been reported in the literature. In this study, high-cholesterol diet (high cholesterol diet, HCD) in rabbits with cholesterol gallstone model, selective COX-2 inhibitor celecoxib (celecoxib, trade name: West Celebrex) immunoglobulin, drill protein there is a certain relationship nucleation factor associated with inflammation in the rabbit bile and serum content changes and observe rabbit gallbladder epithelial degree of inflammation and bile crystallization, and to explore the role of selective COX-2 inhibitors in cholesterol gallstone formation. Called the feet of the dead will be Jun ordinary 24 rabbits (3 to 4 months of age, male or female, weight 2.2 2.gkg) were randomly divided into two groups: control and experimental groups. Ordinary pellet feed feeding 1 week after grouping individual cages in the control group fed a high cholesterol diet of 1.2% in the experimental group on the basis of the 1.2% high cholesterol diet by 10 mg · kg · d 'plus feeding selective COX inhibitor Celebrex. 7 weeks after 12h fasting rapidly fatal Caesarean section, take the cavity 5 ml of venous blood serum was separated by centrifugation and set a 20 ° C freezer storage equipment seized. Simultaneous separation of gallbladder bile duct ligation in the gallbladder neck breaking away from the gallbladder, bile collection into the stone crystallization was observed, and set a 70 ° C refrigerator equipment seized. The gallbladder via rinsed with physiological saline was visually observed after the internal Bo membrane changes. Visual observation of the formation of bile color, Burgundy and the presence or absence of cholesterol crystals the (diameter gt; lIIun, forming a thick sand-like particles). While observing the changes in the thickness of the gallbladder wall and gallbladder anvil film. Gallbladder specimens the the Yi Xue Ying learn Tuo master's degree in less play husband fixed in 10% formalin Fen congregation, the embedded sections by [Le staining,, post, Zhejiang University School of Medicine, Department of Pathology light microscope cholecystitis, with reference to the the Jinggangshan gastritis standard grading. Of normal rabbit serum 2ml, twice with saturated ammonium sulfate precipitation, centrifugation, PH7.04, complex 0.Olmol/LPBS capacity to 2-3ml dialysis three days, plus horseradish peroxidase zomg, stirring to mix. add sodium periodate, stirring was continued together with Sephadex G-25 antigen was concentrated, and set 4OC refrigerator stable antigen-24h, centrifuged, set the volume to 2 ml, was slowly added dropwise a of NaBH4 solution stability reaction, then saturated ammonium sulfate precipitate , centrifugal, double capacity to a 2ml dialysis three days, add equal amount of 60% glycerol mixing, the enzyme-labeled antigen, set 4 ℃ refrigerator and stored. Rabbit bile and serum workers GA, workers GG, Determination of workers gm content using enzyme-linked competitive inhibition assay. Diluted with CBS antibody (goat anti-rabbit ENGINEERING g) coated microtiter plate, 4'C overnight: washed five times with the washing liquid, closed with 15% fetal calf serum at 37 ° C is placed 4Omin; 2 times washed with a washing liquid, and Qiao% fetal calf serum dilution of antigen (rabbit bile and serum) at 37oC placed 2.5h; washed 5 times with washing solution, added with 15% fetal calf serum at 1:100 dilution of the enzyme-labeled antigen at 37 ° C for 2h placed; 5, washed with a washing liquid with TMB (tetramethylbenzidine}

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CLC: > Medicine, health > Internal Medicine > Digestive and abdominal diseases > Liver and gall bladder disease > Gallbladder disease
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