| The first part; to evaluate the viability and energy metabolism of long warm ischemically damaged pancreas during preservation by the UW ( University of Wiscosin) solution cold storage method. The second part: to study the protective effects of L - Arginine ( L - Arg) on reperfusion injury following segmental pancreatic transplantation in canines. The Third part; to investigate the effects of the Astragalus during hypothermia perfusion and preservation of pancreas in-vitro.MethodsThe first part; the pancreas grafts subjected to 30-120 min warm ischemia were preserved by the UW solution cold storage method for 24h. The tissue concentration of adenosine triphosphate ( ATP) and total adenine nucleotides ( TAN ) were determined using high performance liquid chromatography ( HPLC) and the viability of the pancreas graft was tested in the canine model of segmental pancreas autotransplantation. The second part; Segmental pancreatic transplantation transplantation was used. The experimental groups using the L -Arg (200mg/kg) or N - nitro - L - argnine methyl ester ( L - NAME) (10mg/ kg) as an additive agent into the traditional Euro Collin (EC) solution,flushed the isolated pancreas of dogs and stored at 0 ~ 4℃ for 24h, in the meanwhile the group using the EC solution was the control. Segmental pancreatic transplantation transplantation was used. Serum concentration of amylase and lipase were detected. In the pancreas graft, the amount of Nitric oxide (NO) was assessed by Griess reaction method, nitric oxide synthetase ( NOS) was examined with acommercial kit, The expression of inducible nitric oxide synthetase (iNOS) mR-NA was detected by reverse transcription - polymerse chain reaction (RT -PCR) technique , and the activity of myeloperoxidase (MPO) was measured by photometrical method. Histological observation was performed. The Third part: The experimental group using the extractum of Astragalus as an additive agent into the traditional EC solution, flushed the segmental pancreas of canines invitro and stored at 0 ~4C€ for 6h,12h and 24h,in the meanwhile the group using the EC solution was the control. The pancreas cortex were taken for morphology a-nalysis and ultrastructure study from different perfusate used and different time preserved. In addition, autotransplanted these segmental pancreas of preserved 24h in the experimental dogs. Blood sugar (BG) was examined before operation and once a day after pancreas transplantation. Introvenous glouse tolerance test (IVGTT) curve was exeamined a week after pancreas transplantation.Statistical analysis: All data are expressed as mean ± SD. Analysis - of -variance (ANOVA) was used to compare values of the multiple groups. Student t test was used to compare values be twee two groups. Fisher test for comparison of viability.ResultsThe first part: the functional success rates of pancreas grafts of groups after 30 min,60 min,90 min, 120 min of warm ischemia were 100% , 100% , 67.7% ,0% respectively. There was an excellent correlation between the postt-ransplant viability and tissue concentration of ATP and TAN at the end of preservation. The second part: the amount of NO in the L - Arg group was higher than in the control group, while the L -NAME group was lower than control group(p <0. 05). The activitiy of NOS L - Arg group was higher than in the control group, while the L - NAME group was lower than control group (p <0.05). The expression of iNOS mRNA in L - Arg group was higher than in other two groups. The activity of MPO in the L - Arg group was obviously decreasd as compared with in the other groups. The pancrease inflammation was ameliorated when L -Arg was administered, while the pancrease damage was aggravated when L -NAME was administered . The Third part; lightmicroscopic and Electromicro-scopic studies showed that histomorphological changes of pancreas in experimental group were much less than that in control group. The value of BG in experimental group was significantly lower than that in control group ( p < 0. 05). IVGTT curve...
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