| Part 1:The Structure and Function Lesions of Rat Pituitary with Severe Acute PancreatitisObjective:To explore the structure and function lesions of rat pituitary with severe acute pancreatitisMethods:76 SD rats were randomized grouping into 9 groups, each group has 8 rats, the extra 4 rats were grouped into the SAP-12h group. The 9 groups are the blank control group (the N group), the sham-operated group (the A group) and the severe acute pancreatitis model group (the SAP group). The A group and the SAP group were separated into 4 sub-time group, that was the 1h,3h,6h and 12h sub group. We chose the retrograde injection of sodium taurocholic acid into the bile pancreatic duct as the method to make the severe acute pancreatitis model. The operation was under the anesthesia of isoflurane, and the A group was only treated with the flip of pancreas and duodenum. The rats were execute during 12am to lpm, and the exact modeling time and the execute time should be satisfied with the set time of each sub-group. The serum were used to test AMY, LIPA, ALT, Cr and the pituitary hormone GH, ACTH, TSH and FSH. The pancreas tissue were used to measure the pancreas injury condition and the pathological score. The pituitary tissue were used to measure the injury condition and to manufanture the electron microscope slices.Results:Along with the prolonged SAP modeling time, the serum level of AMY, LIPA, ALT and Cr level was elevated significantly (P<0.05). The pancreas tissue slices were found with a obvious change in the edema, hemorrhage, necrosis and inflammatory infiltration, the pathological score changed as the same with a highest score at the 12h (P<0.05). The serum GH level were stable among each group (P> 0.05). The serum ACTH and TSH level of SAP groups were rised fist and then declined with a lower level at 12h when compared with the A group at the same time point (P<0.05). The serum FSH level of SAP groups were continued lower, the 12h serum FSH level was significantly lower when compared with the N group and the A group at the same time point (P<0.05). The pathological slices of pituitary showed the main pathological lesion was hyperaemia, the electron microscope results showed the pituitary cell had a significan endoplasmic reticulum swelling, some mitochondrial swelling and autophagosome and karyopyknosis.Conclusion:There was a pituitary injury in rat with severe acute pancreatitis. The SAP pituitary injury is showed in its functional changes and histomorphology changes.Part 2:The Exact Pathological Injury Form and the Mechanism of Pituitary in Rats with Severe Acute PancreatitisObjective:To explore exact pathological injury form and the mechanism of pituitary in rats with severe acute pancreatitisMethods:60 SD rats were used in this part,24 of those were used in the first phase of this part, they were randomized grouping into 3 groups, these groups were the blank control group (the N group), the 12h sham-operated group (the A-12h group) and the 12h severe acute pancreatitis model group (the SAP-12h group) with 8 rats in each group. The other 36 rats were used in the second phase of this part, they were randomized grouping into 9 groups, these groups were the blank control group (the N group), the sham-operated group (the A group) and the severe acute pancreatitis model group (the SAP group), and the A group and the SAP group were separated into 4 sub-time group, that was the lh,3h,6h and 12h sub group. Each of those groups has 4 rats. We chose the retrograde injection of sodium taurocholic acid into the bile pancreatic duct as the method to make the severe acute pancreatitis model. The operation was under the anesthesia of isoflurane, and the A group was only treated with the flip of pancreas and duodenum. The rats were execute during 12am to lpm, and the exact modeling time and the execute time should be satisfied with the set time of each sub-group. The pituitary tissues in the first phase were used to detect the expression level of Caspase 3,8,9,12 and CHOP, and the second phase’s tissues were used to measure the injury condition by electron microscope.Results:The SAP rats’s pituitary has a high expression level of Caspase 3,8,9,12 and CHOP when compared with the N group and A group (P<0.05). When we compared the expression level of Caspase 3,8,9,12 and CHOP of pituitary in SAP rats, we found the level of CHOP is significantly higher than the other molecules (P< 0.05). Along with the prolonged SAP modeling time, the electron microscope results showed the pituitary cell had a significan endoplasmic reticulum swelling, this change was found in different cells of pituitary, and the endoplasmic reticulum change was even worse along the time of SAP.Conclusion:The main pathological injury form of SAP rat’s pituitary is endoplasmic reticulum stress and its induced cell change. The way of unfolded protein response to CHOP to apoptosis maybe the mechanism of the SAP pituitary injury.Part 3:The Interfere Effect of 4-PBA on Pituitary Injury of Rat with Severe Acute Pancreatitis and Its MechanismObjective:To explore the interfere effect of 4-PBA on pituitary injury of rat with severe acute pancreatitis and its mechanismMethods:36 SD rats were used in this part, they were randomized grouping into 3 groups, each group has 12 rats. The 3 groups are the sham-operated group (the A group), the severe acute pancreatitis model group (the SAP group) and the 4-PBA interfere group (the 4-PBA group). We chose the retrograde injection of sodium taurocholic acid into the bile pancreatic duct as the method to make the severe acute pancreatitis model. The operation was under the anesthesia of isoflurane, and the A group was only treated with the flip of pancreas and duodenum. The rats of 4-PBA group were treated with the intraperitoneal injection of 4-PBA at the concentration of 10mg/mL, the dose was 50mg/kg of the body weight. The treatment of 4-PBA injection was started three days before the SAP modeling. The rats were execute during 12am to lpm, and the exact modeling time were set 12 hours before the execution. The serum were gathered to test the serum AMY, LIPA, ALT, Cr and the serum GH, ACTH, TSH and FSH, the pancreas tissue were used to measure the pancreats pathological changes, the pituitary tissue were gathered too, the 8 of them were used to measure the expression level of CHOP, Bip and ORP 150, the other 4 pituitary were used to confirm the electron microscope changes.Results:By using of 4-PBA, the rats’ pancreatitis were controlled, the serum AMY, LIPA, ALT, Cr level were lower than the SAP rats, the pancreas pathological change were improved either, the pathological of pancreas is lower in 4-PBA group when compared with the SAP group (P<0.05). We also found that the serum ACTH, THS, FSH level of 4-PBA group were higher than the SAP group (P<0.05). When compared with the SAP group, the 4-PBA group pituitary has a lower expression of CHOP, Bip, ORP 150 (P<0.05), the pathological change is also improved. By view the electron microscope slices of each groups, we found the endoplasmic reticulum swelling were significantly improved by interfere of 4-PBA.Conclusion:4-PBA can protect the pituitary in rats with severe acute pancreatitis by inhibit the unfolded protein response, the unfolded protein response to CHOP pathway of endoplasmic reticulum stress may paly an important part in 4-PBA’s protection process on SAP pituitary injury. |
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