| ObjectiveTo establish a model of alcoholic chronic pancreatitis in rats by ethanoland lipopolysaccharides(LPS, endotoxin). To observe TLR4(toll-likereceptor4)pathway activation status，analysis the relationship of TLR4pathway activation and alcoholic chronic pancreatitis.To test the bloodendotoxin level、the oxidative stress of pancreas in each group, to makepreliminary research on the mechanism of TLR4pathway activation.Methods1In order to establish a rat model of alcoholic chronic pancreatitis,twenty-four male SD rats were randomly divided into4groups: controlgroup, alcohol group, lipopolysaccharides (LPS) group and LPS combinedalcohol group(combined group). All the rats were fed with the same food.Rats in control group and LPS group were fed with water, and the otherswere fed with25%alcohol. After twelve weeks, the rats in model group andLPS group were intraperitoneal injected with LPS （2mg/Kg），once aweek, for4times，at the same time，control group and alcohol group wereintraperitoneal injected an equal volume of saline solution. All rats werekilled1week after the last injection.2To study the influence of alcohol combined with lipopolysaccharideon the blood endotoxin level, each time after endotoxin injection for1daysand6days took the rat tail blood of each group to determination theendotoxin concentration by azo reagent method.3To study the influence of alcohol combined with lipopolysaccharideon pancreatic toll-like receptor4(TLR4) pathways, the expression of leukocyte differentiation antigen14(CD14)mRNA、TLR4mRNA、tumornecrosis factor-α(TNF-α)mRNA in the pancreas of each group wereobserved by RT-PCR，and the expression of CD14protein、TLR4protein、TNF-α protein in the pancreas was determinated by immunohistochemistry.4To study the effect of alcohol combined with lipopolysaccharide onpancreatic antioxidation, xanthine oxidase method was used for thedetermination of the content of superoxide dismutas(SOD),thiobarbituricmethod for the determination of malonaldehyde (MDA)content in eachgroup.Results1Compared with other groups, the inflammatory cell infiltration andcollagen increasing were observed in combined group.2Compared with other groups, Every one days after injection ofendotoxin，blood endotoxin level in model group was significantly higherthan that of the control group and lipopolysaccharide group(P <0.01),higher than alcohol group (P <0.05), The endotoxin level of alcohol groupwas higher than that of the control group and lipopolysaccharide group (P <0.05). Every6days after injection of endotoxin，the endotoxin level ofmodel group and alcohol group were higher than those of the control groupand lipopolysaccharide group (P <0.05), no difference between the modelgroup and alcohol group.3Compared with alcohol group，in combined group the expression ofCD14protein, TLR4protein, TNF-α protein and CD14mRNA, TLR4mRNA, TNF-a mRNA was increased(P <0.05)，Compared with controlgroup and LPS group，in combined group the expression of CD14protein,protein TLR4, TNF-α protein and CD14mRNA, TLR4mRNA, TNF-αmRNA was significantly increased. 4Compared with other groups，in combined group the expression ofMDA was increased in pancreatic tissue，however, the expression of SODwas decreased in pancreatic tissue (P <0.05)。Conclusion1Combined with long-term ethanol intaking, intraperitoneal injectionof LPS can induce rat chronic pancreatitis.2Combined with long-term ethanol intaking, intraperitoneal injectionof LPS can significantly increase blood endotoxin concentration, long-termalcohol intaking may also increase blood endotoxin concentration, thepathogenesis of alcoholic chronic pancreatitis may be related to bloodendotoxin concentration.3Combined with long-term ethanol intaking, intraperitoneal injectionof LPS can effectively promote the expression of CD14, TLR4and TNF-α,long-term alcohol intake can also increase the expression of CD14, TLR4and TNF-α， the pathogenesis of alcoholic chronic pancreatitis may berelated to highly activation of TLR4pathway.4Combined with long-term ethanol intaking, intraperitoneal injectionof LPS can effectively promote the MDA expression, inhibite SODexpression, alcoholic chronic pancreatitis pathogenesis may be associatedwith pancreatic oxidative stress.. |
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