The Mechanism Of Sandostatin For Pancreat-cell Apoptosis Of Severe Acute Pancreatitis And The Effect Of Sandostatin For Peripheral Blood Neutrophil Apoptosis In Rats

Objective This experiment study the effect of Sandostatin for peripheral blood neutrophil apoptosis in severe acute pancreatitis in rats; To study the mechanism of Sandostatin for pancreat-cell apoptosis of severe acute pancreatitis and the effect of Apoptosis-related gene bax, bcl-2 in rats.Methods: Forty-eight male SD rates were divided into 4 groups randomly: severe acute pancreatitis (SAP) group, Sandostatin-treated (SDT) group, Ulinastatin-treated (UTI) group, sham operated (SO) group. Each group was divided into 4-hour subset and 14-hour subset, 6 rats in each subset. After successful animal model production,the 4-hour subset was killed after 4 hours and the 14-hour subset was killed after 14 hours. And then draws blood 6 ml from the rat heart, 3 ml of which is reserved for detection of neutrophil apoptosis, and another of which is sended the biochemistry room to check serum amylase. Purification neutrophil sends to test neutrophil apoptosis by flow cytometry. Fetch fresh pancreatic tissue being put in 4% paraformaldehyde solution. Part of pancreatic tissue is changed under light microscope by HE staining; Part of pancreatic tissue is detected bax, bcl-2 protein expression by immunohistochemistry staining; Part of pancreatic tissue is tested the Apoptotic Index dy TUNEL staining.Results: 1.The level of AMY in SDT4,UTI₄ and SO₄ is significantly lower than the one in SAP4 (P <0.05),the same as the 14-hour subgroups in the statistical sense. The level of AMY in SO₄ is significantly lower than the one in SDT4, UTI₄ was statistically significant (P <0.05), the same as the 14-hour subgroups in the statistical sense. The others group is no significant difference in the level AMY. And there is no significant difference between each subgroup. 2. There was a massive ascites mixed the blood in UTI, SAP, SDT Groups, Congestive swelling of the pancreas, working closely with the surrounding tissue adhesions, or even a small black necrotic. There were minorly mesenteric edema in SO. There can been seen inflammatory cell infiltration, and the gland swelling, spotty, and fibrinoid necrosis of large, structural disorder, typical morphological changes of apoptosis cells in UTI, SAP, SDT Groups. There no obvious abnormalities in SO. 3. Immunohistochemical staining showed the bax gene expression in pancreatic acinar cell cytoplasm. Compared with SO₄(1.93±1.2),the bax gene expression had a significant lower level than that in SDT4(14.16±2.54),UTI₄ (11.07±2.57)and SAP4(11.25±2.33) (P<0.05),the same as 14-hour subgroup in the statistical sense (P<0.05). Each subgroups of SDT (4, 14) had significant higher level of the bax gene expression than that in UTI, SAP and SO (P<0.05). There was no significant statistical significance between the others. Every 4-hour subgroups and 14-hour subgroups didn't have significant statistical significance in each other. 4. Immunohistochemical staining showed the bcl-2 gene expression in pancreatic ductal cells. Compared with SO₄(2.82±1.80), the bcl-2 gene expression had significant lower level than that in SDT4(17.69±2.27),UTI4(17.23±3.65)and SAP4(13.84±2.38) (P<0.05) the same as 14-hour subgroup in the statistical sense (P<0.05). Each subgroups of UTI (4, 14) had significant higher level of the bcl-2 gene expression than that in SAP (P<0.05), but didn't have significant statistical significance with SDT. Each subgroups of SDT (4, 14) had significant higher level of the bcl-2 gene expression than that in SAP (P<0.05). Every 4-hour subgroups and 14-hour subgroups didn't have significant statistical significance in each other. 5. The Results of TUNEL showed that SDT4(6.03±0.93) had significant higher level of the pancreatic apoptosis index than that in SAP4(4.73±1.07),SO₄(1.69±1.39),UTI4(4.68±1.06) (P<0.05).The number of the pancreatic apoptosis index didn't have significant statistical significance in UTI4 and SAP4. SO₄ had significant lower level of the pancreatic apoptosis index than that in erery 4-hour subgroups (P<0.05),the same as the 14-hour subgroup in the statistical sense (P<0.05). SDT4 (6.03±0.93) had significant higher level of the pancreatic apoptosis index than that in SDT₁₄ (6.12±1.15)(P<0.05), and the others subgroups have shown the same changes, but they didn't have the statistical significance. 6. Neutrophil apoptosis detection: SDT4(5.53±2.19) had significant higher level of the rate of neutrophil apoptosis than that in SO₄(1.50±0.76), SAP4(2.85±1.27)(P<0.05), and had no statistical significance with UTI4(7.06±1.86). SDT₁₄(3.59±1.04) had significant higher level of the rate of neutrophil apoptosis than that in SO14(1.32±0.53), SAP14(1.90±0.89)(P<0.05), and had significant lower level than that UTI₁₄ (5.22±1.58)(P<0.05). UTI4 had significant higher level of the rate of neutrophil apoptosis than that in SO₄ and SAP4 (P<0.05). UTI₁₄ had significant higher level of the rate of neutrophil apoptosis than that in SO14, SAP14 and SDT₁₄ (P<0.05). SDT4 had significant higher level of the rate of neutrophil apoptosis than that in SDT₁₄, the same as UTI-group. SAP4 and SAP14 didn't have the statistical significance the same as SO-group.Conclusions: 1.In the early process of SAP Sandostatin can induce pancreatic Cell Apoptosis, which can relieve the pancreatic necrosis. Ulinastatin which can inhibit trypsin don't play significantly role in promoting apoptosis of pancreatic cells. The bax gene express in pancreatic acinar cell cytoplasm. Sandostatin can increase bax gene expression. The bcl-2 gene express in pancreatic ductal cells. Sandostatin and Ulinastatin can increase bcl-2 gene expression.2. Sandostatin and Ulinastatin can improve the lagging role in neutrophil apoptosis. But with the horary extension, Ulinastatin in prompting neutrophil apoptosis have the more obvious effects than Sandostatin.