| Objective:To study the effect and the possible mechanisms of inflammation signal transduction pathways of severe acute pancreatitis rats by detecting the AMY,TNF-a,IL-1βin blood serum,observing changes of Panreatic tissues through light microscope and the expressions of TLR4,NF-ΚB in pancreatic tissues. Method:120 wister rats,female and male in half, with the weight of 220-260g,were randomly divided into 5 groups, including sham operation group (J group),severe acute pancreatitis group (M group),somatostatin group (S group),YI DAN extrinsic applyment group (Y group),YI DAN Combined Modality Therapy (Z group).Rats in J group whose pancreas were just flipped and striked gently with nothing injected 0.5% sodium taurocholate in 1.0 ml/kg were retrograde inject into pancreatic duct of the rats in M group,S group, Y group,Z group reduce SAP.After operation,rats were deprived of food with free access to water.Rats in S group,50 ug/kg subcutaneous injections respectively before duplicating model half a hour.Rats in Y group, ex-apply YI DAN on epigastric region of rats, changing dressings everyday,7 times for one course of treatment;Rats in Z group,50 ug/kg subcutaneous injections respectively before duplicating model half a hour ex-apply YI DAN on epigastric region of rats, change dressings everyday,7 times for one course of treatment.6 rats of every group were observed in 3h,12h,72h,2W.And then 6 rats of every group were humanely killed at 3h,12h,72h,2W after observing changing of severe acute panreatitis rats.Compared of pancreatic injury in every group through light microscope.Blood serum was obainted for detecting the rats'AMY.TNF-αand IL-1βwere detected by ELISA. TLR4 and NF-ΚB in Panreatic tissues were semiquantitative detected by immunohistochemical methods.All the results were analysed by Anova with SPSS 16.0.Result:①Pathological change of pancreatic tissues:In general morphous,the observation through light microscope:the structure of pancreatic of J group were normal in general.In M group, the pancreatic lobule filled with edema, phlegmonosis cell infiltrated obviously, with time continuing,on the edge of crypt expand gradually with phlegmonosis cell.pancreas parenchyma Inflammation>50%,pancreas parenchyma hemorrhage>50%,pancreatic crypt appeared spotty necrosis and lobule deletion at 12h extreme obviously.Those also appeared in S group and Y group,with time continuing,the pancreatic tissues were better.The Z group of pancreatic lobule filled with edema,phlegmonosis cell infiltrated obviously, with time continuing,on the edge of crypt expand gradually with phlegmonosis cell, pancreas parenchyma Inflammation>50%,pancreas parenchyma hemorrhage>50%.All those appeared at 3h extreme obviously.With time continuing, the edema,Inflammation,hemorrhage,riecrosis of pancreatic tissues were alleviated obviously.②Pathological change of pancreatic tissues and pathological score:the pathological score of J group had no obvious differences in different times (P>0.05).The pathological score of M group,S group and Y group was higher than J group(P<0.01).With time continuing, the pathological score of M group,S group and Y group was higher gradually,24h is the Peak time.Z group was lower than M group,S group and Y group in different times (P<0.01).(2) AMY (U/L):The level of AMY in blood serum of J group was lowest.The concentrations of AMY in blood serum were increased gradually after duplicating SAP model,they were 12h is the Peak time,72h reduce gradually,2W was lowest. The AMY of activity variance regularity of Z group in different times was identical to M group,S group and Y group, while the activity variance of Z group in different times was lower than M group,S group and Y group (P<0.01). (3) TNF-α(pg/ml):Compared with the J group,higher level TNF-αof M group,S group,Y group and Z group in blood serum were found in different times (P<0.01).Moreover, Compared with the M group,S group and Y group, the level TNF-αof Z group decrease obviously(P<0.05 or P<0.01).(4) IL-1β(pg/ml):Compared with the J group,higher level IL-1βof M group,S group,Y group and Z group in blood serum were found in different times (P<0.01).Moreover, Compared with the M group,S group and Y group, the level IL-1βof Z group decrease obviously(P<0.01).(5) Expressions of TLR4 protein in pancreatic tissues:The results of immunohistochemical method about TLR4 protein in pancreatic tissues were increased expressing by IOD.The activation of TLR4 in J group was lowest.The level of TLR4 protein was activated swiftly and secreted within 3h after duplicating SAP model,reached peak value at 12h,and then fell after 72h to normal level. The TLR4 of activity variance regularity of Z group in different times was identical to M group,S group and Y group, while the activity variance of Z group in different times was lower than M group,S group and Y group (P<0.01). (6) Expressions of NF-ΚB protein in pancreatic tissues:The results of immunohistochemical method about NF-ΚB protein in pancreatic tissues were increased expressing by IOD. The activation of NF-ΚB in J group was lowest. The level of NF-K B protein was activated swiftly and secreted within 3h after duplicating SAP model,meanwile,reached peak value at 3h,also reached peak value at 12h,72h reduce gradually,and then 2W was lowest.The NF-ΚB of activity variance regularity of Z group in different times was identical to M group,S group and Y group, while the activity variance of Z group in different times was lower than M group,S group and Y group (P<0.01). Conclusion:Among the pathological change of pancreatic tissues,the level of AMY,TNF-a and IL-1βin blood serum,the expression of TLR4 and NF-KB in Panreatic tissues,all those index ameliorate obviously after YI DAN combined modality therapy of rats with severe acute panreatitis (P<0.05);Horizontal movement and vertical movement of Z group increase obviously at anaphase (P<0.05).TLR4 participate in the signal transduction of inflammatory cell of SAP.YI DAN combined modality therapy can down regulate the expression of TLR4 level,also can inhibit NF-ΚB activation,suppress the level of Inflammatory cytokines,and finally attenuate the severity of SAP,which may be one of the mechanism of these drugs on the influence of on the signal transduction of inflammatory cell of the SAP rats.Meanwhile,which may be consensual consequence after releasing Inflammation of the SAP rats.
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