The Study Of Effect Of Anti-P-selectin Monoclonal Antibody In Preventing The Ischemia/Reperfusion Injury Of Rat Pancreas

Transplantation of pancreas is one of the fundamental measures to treat diabetes mellitus. It is necessary to cut off and preserve donor pancreas before transplantation. Therefore, ischemia reperfusion injury is unavoidable. Preventing reperfusion injury is the key step for the survival of donor pancreas and is also the hot spot of the present research. The mechanism of ischemia re reperfusion injury has not been completely clarified, yet it is known that it is closely correlated with the expression of inflammatory factors. The activation of P - selection is an important link to switch on many inflammatory factors. The present research lays emphasis on the block of some kind of inflammatory medium and the effect is limited. In our study, anti - Ps monoclonal antibody is used to block the effect of Ps, thus blocking the initial step of inflammation, inhibiting the cascade reaction of inflammatory factors and preventing the reperfusion injury of donor pancreas basically. The effective prevention of ischemia/reperfusion injury means the reduction of the complications after transplantation of pancreas. This has important sense for the survival and functions of the transplanted pancreas.Material1. Examination animal;Male Wistar rat, 200 ± 20g, provided by animal center of China medical university.2. Apparatus and reagent; Microscope surgery instrument RefrigeratorySlice machineOLYMPUS Microscope and image analysis instrument Ps monoclonal antibody, purchased SuZhou medical college Ps, ICAM - 1 ,TNF - a monoclonal antibody and kit, purchased BOSTER biological technology ltd.Method1. Sample collection;This examination adopts pancreas ischemia/reperfusion model, divide all rats into control group, ischemia/reperfusion group and Ps monoclonal antibody managed group, every group include 15 rats, and all groups are divided into 3 parts according to 3 period of time. After opening the abdomen, find the subsplenic arterial, we do not deal with the control group; Ischemia reperfusion group shut off 30 min before reperfusion, get material at the time of 15,30,60 min; Exam group is injected Ps monoclonal antibody from dorsal vein of penis 5 min before reperfusion. The materials are fixed by 10% formaldehyde confected by PBS.2. Observed index; make the sample paraffin slice.2.1 HE dyeing; dye all tissue and observe tissue structure.2.2 Immunohistochemistry: show Ps,ICAM - 1,TNF - a express condition, put the slide on the microscopic image analysis machine and collect data.3. Statistical analysis by SPSS 10.0 system.Result1. Tissue configuration:There were not distinct injury in pancreatic structure of both control group and exam group, while pancreatic structure of ischemia/ reperfusion group change relative distinct and aggravated with the increase of reperfusion time.2. Immunohistochemistry:No significant difference was found between control group and exam group in Ps JC AM - 1 ,TNF - a ( P > 0.05 ). But, on the contrary, significant difference was found among the ischemia reperfusion group,control group and exam group ( P < 0.05 ). But there was significant difference in ischemia reperfusion group at different phase.ConclusionAnti - P - selectin monoclonal antibody has protection effect on ischemia /reperfusion injury. P - selectin,ICAM - 1 and TNF - a expressed increased lonely with time, Ps maybe the initial factor of ischemia reperfusion injury.