Effect Of Kupffer Cell Inhibition On Severe Acute Pancreatitis

Objective: To investigate the effect and mechanism of Kupffer cell inhibition on severe acute pancreatitis.Methods: Healthy female Wistar rats were divided at random into three groups: Sham operation group (SO group), severe acute pancreatitis group (SAP group) and gadolinium chloride group (GdCl₃ group). Severe acute pancreatitis was induced by retrograde injection of 5% sodium taurocholate (0.1ml/100g) in common bile duct in SAP group rats. One day before the operation the gadolinium chloride solution (20mg/kg) was injected into the tail vein in GdCl₃ group rats. The operation which induced severe acute pancreatitis model of GdCl₃ group rats was same to SAP group rats. The pancreas and duodenum were gently dragged in SO group rats. The rats of each group were killed 24 hours after manipulation. The experiment included three parts. 1,Experiment one: (1) To detect endotoxin,alanine aminotransferase (ALT),amylase (AMS),tumor necrosis factor (TNF-α),nitrogen monoxide (NO),malondialdehyde (MDA) in blood and liver homogenate tumor necrosis factor (TNF-α) content. (2) HE stain, assisted with light microscope to observe pathological changes of pancreas and liver. (3) By using flow cytometer to estimate the liver cell DNA level and to judge the occurrence of apoptosis. What's more, we could calculate the liver cell apoptosis rate. 2,Experiment two: Before the execution, India ink 1:3 diluted with germ free physiological saline (0.25ml/100g) was injected into the vail vein in each group rats. (1) To calculate phagocytic index k and its correction a. (2) HE stain, assisted with high power light microscope to observe Kupffer cell which phagocytosed India ink particles and to count the cells. 3,Experiment three: To detect endotoxin,endothelin (ET-1),nitrogen monoxide (NO),thromboxane B₂ (TXB₂),6-Keto-prostaglandin1α(6-Keto-PGF_1α) in blood content. We could also calculate ET-1/NO and TXB₂/6-Keto-PGF_1α .Results: 1,Experiment one: (1)The levels of endotoxin,ALT,AMS,TNF-α,NO,MDA in blood and liver homogenate TNF-αin SAP group significantly increased (vs SO group, p<0.01). The level of endotoxin in blood in GdCl₃ group significantly increased (vs SAP group, p<0.01). The levels of ALT,TNF-α,NO,MDA in blood and liver homogenate TNF-αin GdCl₃ group significantly decreased (vs SAP group, p<0.01,0.01,0.05,0.01 and 0.01). The level of AMS in blood in GdCl₃ group did not apparently change vs SAP group. (2) In SO group rats'pancreas: Glandular lobule structure was normal. Acinus and duct formation were integrated. Cell constitution was natural. In SAP group rats'pancreas: Massive necrosis,...