| Objective retrograde Cholangiopancreatic duct injection of 5% sodium taurocholate caused injury model in rats SAP, monitoring and testing of the physiological and laboratory parameters that associated with SIRS, used to clear the mechanism of early hyperlipidemic increase pancreatitis.Methods Clean SD (SPrague Dawley) male rats were 90, were randomly divided into normal control group(N):Basal diet for 8 weeks, the day of surgery Measuring rectal temperature, monitoring heart and respiratory rate after the success of anesthetized rats, then only flip the duodenum and pancreas after the median abdominal incision, free water after they palinesthesia. Hyperlipidemia control group(HLN): High fat diet for 8 weeks to modeling HTG model, Method of operation at the same modeling with normal control group. Severe acute pancreatitis(SAP) group: Basal diet for 8 weeks, Measuring rectal temperature, monitoring heart rate and respiratory rate after the success of anesthetized rats, retrograde injection of electronic micro-pump of 5% taurocholic acid (0.1ml/100g, 0.1ml/min) to establish animal model of SAP mouse. Hyperlipidemic Severe acute pancreatitis group (HLSAP): High fat diet for 8 weeks to modeling HTG model, Method of operation at the same modeling with SAP group. Each group was divided into three experimental time points, total of 12 groups. Rectal temperature measurement, monitoring heart and respiratory rate again after Each period rats were re-anesthetized, then abdominal cavity was opened along the original incision. Collection, measurement of ascites, collected 1ml abdominal aorta blood for arterial blood gas analysis, remaining after centrifugation, retain the serum for Measurement of serum amylase, serum lipase, IL-10, IL-6 and TNF, etc. Conventional dehydration of pancreatic tissue, embedded in paraffin, paraffin routine HE staining, pathological scoring pancreas tissue with microscopic.Results Rats in each group, none died during the feeding period, the sham operation group model no death also. HLSAP 12 hours group with four deaths, 6 hours group with two deaths; 2 hours was no death. SAP12 hours group, there are two death within 12 hours, 6 hours group with 2 deaths, 2 hours was no death. After 8 weeks high fat diet fed rats weighing between 330g-486g, compared with normal diet group 223g-326g increased significantly, the difference was significant.high-fat diet group Serum TG,TC, respectively1.63±0.51mmol/L, 1.34±0.62mmol/L, compared with normal control group TG 0.54±0.24mmol/L, TC0.57±0.37mmol/L increased significantly, compared with two sets of data P<0.05, significant difference.Gross pathological changes of pancreas: Eye view of the control group at each time point pancreas did not change significantly, the appearance of normal, was pale pink, uniform color, glossiness good. SAP group: Modeling at different time points after re-laparotomy found: 2h group showed a small amount of reddish abdominal ascites, congestion and edema of pancreatic tissue began, there were several spots hemorrhage; texture is still soft. 6h group of ascites increased, the color was red or dark red bloody ascites, flaky pancreatic necrosis, gastric omentum, mesentery at the small quantity of yellow-white punctate calcification. 12h group the color of ascites was further deepen, but the amount no significant increase, pancreatic texture hard, see large black necrotic foci, some showed cystic degeneration of pancreatic tissue, omentum, superior mesenteric see multiple calcification. HLSAP group: compared with SAP group 2 hours and 6 hours time point was no significantly different in gross pathological changes, 12-hour group compared with SAP group have bigger necrotic area.Microscopic pathological changes of pancreas: Structural integrity of the control group glands, glandular epithelium showed no abnormal changes, some rats can be seen in varying degrees of leaves gap widened, occasionally can be seen inflammatory cells. HL group: Lipid vacuoles were seen in some cells, no significant bleeding and necrosis, remaining the same as the control group. SAP in each group showed different degrees of change. 2 hour group: Pancreatic interstitial edema, leakage of red blood cells, acinar cell edema, focal necrosis, part of the gland duct ectasia. 6 hours group, Edema, hemorrhage and necrosis increased, infiltration of inflammatory cells increased. 12 hours group: There was no structure of a large necrotic area and inflammatory cell infiltration. compared with SAP group HLSAP group 2 hours and 6 hours time the pathological changes and pathological score was no significant difference, But 12hours group compared with SAP group have more serious necrosis, Lipid vacuoles were seen in some cells and a significant difference between the pathological score.SIRS complicated situation: HLSAP 2 hours group 3 rats can be diagnosed as SIRS. 6 hours group of 5, 12 hours group all diagnosed as SIRS; 2 hours after a normal diet comparison group without SIRS, 6 hour group had 3, 12 hours group of 5.Serum IL-6, TNF-αand IL-10 changes: After the modeling HLSAP and SAP group serum IL-6, TNF-αconcentration started to increase in the time of 2 hours. Compared with the respective control group, significant difference (P <0.05). With the modeling time extended serum IL-6, TNF-αlevels increased gradually, to 12 hours after the model reached the highest value. HLSAP6h serum TNF-α, HLSAP12h serum IL-6, TNF-αat he same time compared with SAP group were significantly different (P <0.05). HLSAP group and SAP group increased serum concentrations of IL-10, With their respective control group, there was significant difference (P <0.05). HLSAP 2h group was higher than the same time point SAP group, the difference was significant (P <0.05), HLSAP12h group was lower than the same time with the SAP group, the difference was significant (P <0.05) Changes of biochemical markers in serum: HLSAP group and SAP serum amylase and lipase concentration Increase, compared with their respective control group, there was significant difference (P <0.05). HLSAP6h serum amylase and lipase levels lower than the same time point SAP group, the difference was significant (P <0.05). HLSAP group and SAP group had the higher blood glucose, SAP group 6 hours, HLSAP group 6 hours and 12 hours compared with the control group, there was significant difference (P <0.05), with the modeling time Extended,blood sugar Gradually have the increasing concentration trend, HLSAP 12h group and the SAP group compared with the same time there was significant difference (P <0.05). HLSAP group and SAP serum ionized calcium were lower compared with their respective control group, there was significant difference (P <0.05). Concentration decreased gradually with the modeling time extended. HLSAP6h, 12h group of calcium were significantly lower than the same time point of SAP group, the difference was significant (P <0.05). HLSAP 2h and 12 group compared with SAP group in the same time there was significant difference (P <0.05)。Changes in the phase of ascites: sham group at each time point occasional have small reddish abdominal ascites , There was red to red turbid ascites after abdominal cavity was opened,With the time extended turbidity And the amount of ascites increased. 6 hours group have the largest amount of ascites, compared with SAP group there was significant difference (P <0.05).SAP group received laparotomy also have red or dark red turbid ascites, 6h and 12h group changed little amount of ascites.Conclusion 1 SD rats by giving high-fat diet for 8 weeks retrograde cholangiopancreatography combined with injection of 5% sodium taurocholate solution can be successfully induced hyperlipidemia in rats with severe acute pancreatitis model, which pathogenesis and pathological similarities with human diseases, Has induced a high success rate, feasibility and good features. 2 Heavier fat SAP group showed earlier SIRS, making it a heavier early indicator of pancreatitis, clinical attention on the need for drug intervention in early. |
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