| BackgroundLiver ischemia-reperfusion(I/R) injury has been an important impaired factor which cause functional lesion in hepatic trauma,resection,transplantation and the treatment of multiple organ failure.The damage mechanism and the protective measure have been generally investigated.Ischemia-reperfusion injury is a multifactorial procedure.The production of active oxygen,the activation of Kupffer cell,lymphocyte and neutrophil are included.A series of harmful reaction are evoked which cause inflammatory reaction and apoptosis.The expression of many cytokines and endocrines such as nitrogen monoxidum(NO),heat-shock protein(HSP), endothelin(ET),hypoxia-induce factor-â… (HIF-â… ),insulin-like factor(IGF-â… ) are changed in the I/R injury.Among all these factors,the effect of IGF-â… has had been more and more investigated.IGF is a group of polypeptide which has insulin-like anabolic action and growth promotion action.An important role is played by them in histiocyte multiplication,differentiation and in inhibiting apoptosis.Recently, IGF-â… has had been paid more attention to because of its abnormal expression and potential effect in liver trauma,liver surgery and multiple organ failure.However,the relationship between IGF-â… and hepatic I/R injury is not elucidated,so do the space-time distribution and significance in hepatic I/R injury.Radix salviae miltiorrhizae is a kind of traditional Chinese medicine.It has been showed the protective effect during I/R injury to some organs.The mechanism includes two aspects.First of all,RSM is able to improve the micro-circumfusion and has an anti-ox radical effect.Secondly,RSM is able to reduce the level of[Ca2+]i and cell damage caused by calcium overload.In formal research,the effect of radix salviae miltiorrhizae on cytosolic free calciumion concentration[(Ca2+)i](nM) in isolated hepatocytes in rats and patients with ischemia reperfusion by microflurometry using fluorescent[Ca2+]i indicator Fura-2/AM has been studied. Changes of lipid peroxide free radical(ROO) signal ranges with in the liver tissue by ESR technique and those of hepatocellular ultrastructure by electron microscope were also observed.The results showed that RSM reduced the levels of[Ca2+]i and ROO·Ymax(mm) ESR signal rangs.RSM had an effect on protecting hepatocytes against ischemia/reperfusion injury as a useful blocker of receptor-operated calcium channels.However,it is not reported that whether these protective effects have any potential association with IGF-â… .Our subject is based on the formal research including two parts to study the effect of RSM on different period's ischemia-reperfusion.In part one,the rat hepatic I/R model was built to observe the expression of IGF-â… and Caspase-3 during different I/R periods and to discuss the effectiveness significance.In part two,by compare the different expression significance of IGF-â… and Caspase-3 between RSM group and no-treatment group,the time-space distribution and effectiveness significance were aim to know.By which a new prevention and cure method was researched. ObjectiveThe rat hepatic I/R model was built to observe the expression of IGF-â… and Caspase-3 during different I/R periods and to discuss the effectiveness significance.Materials and Methods1.75 healthy SD rats were randomly divided into 3 group including normal(N) group(n=25),sham operation(SO) group(n=25) and ischemia-reperfusion group(I/R) group(n=25) Each group was divided into 5 subgroup according to different reperfusion periods(0h,3h,12h,24h,72h).There were 5 rats in each subgroup.2.The rats were been fasting for 12 hours preoperation and were free to drink. Routine preoperational preparation was done.Liver tissues were directly taken for N group.Hepatic portal was dissected only for SO group.In I/R group,non-damage artery forceps was placed onto hepatoduodenum ligament for 45min.The liver tissues were taken in different time(0h,3h,12h,24h,72h).3.IGF-â… and Caspase-3 immunohistochemistry test was progressed on instruction.The ALT,AST levels were tested by auto-biochemistry instrument.The pathomorphological change was observed by light microscope and electron microscope.4.SPSS13.0 was used to do analysis.The method is repeated measured data analysis. ResultThe expressions of IGF-â… in N group and SO group have no significant difference.After ischemia 45min and reperfused,the expression of IGF-â… in liver decreased a little at 0h and significantly at 3h.It reached to the lowest point at 12h and backswing at 24h.The expressions of Caspase-3 in N group and SO group have no significant difference.In I/R group,the expression of Caspase-3 strengthened.It reached to the peak at 24h and was back to the baseline at 72h.After reperfusion,the serum ALT,AST levels were higher than N group and SO group at 0h,3h,12h,24h and was back to baseline at 72h.There was no change of organelles in N group and SO group.Significant damage of organelles in I/R group displayed by light microscope and electronic microscope.The damage is severest at 12h～24h.ConclusionThe expression of IGF-â… has a close relationship to liver function.And this effect has space specificity,that is,the RSM regulate IGF-â… expressing intense in central veins while weak in parenchymatous tissue.As an endogenous protective factor,IGF-â… may has an antagonistic effect to functional lesion by I/R injury. ObjectiveThe rats were divides into ischemia-reperfusion no-treatment group,sham operation group,normal group and RSM precondition group to observe the expression of IGF-â… and Caspase-3 and to investigate the influence of RSM on IGF-â… expression in different periods hepatic ischemia/reperfusion.materials and Methods1.100 healthy SD rats were randomly divided into 4 group including normal(N) group(n=25),sham operation(SO) group(n=25),no-treatment(NT) group(n=25) and Radix Salvia Miltiorrhiza(RSM) group(n=25).Each group was divided into 5 subgroup according to different reperfusion time(0h,3h,12h,24h,72h).There were 5 rats in each subgroup.2.The rats were been fasting for 12 hours preoperation and were free to drink. routine preoperational preparation was done.Hepatic portal was dissected only for SO group.In NT group,non-damage artery forceps was placed onto hepatoduodenum ligament for 45min.the RSM group was built by intravenous injection RSM before I/R. The liver tissues were collected according to different periods(0h,3h,12h,24h,72h).3.IGF-â… and Caspase-3 immunohistochemistry test was progressed on instruction.The ALT,AST level were tested by auto-biochemistry instrument.The path morphological change was observed by light microscope. 4.SPSS13.0 was used to do analysis.The method is repeated measured data analysis.ResultThe expression lever of IGF-â… with the treatment of RSM was higher at 3h,12h and 24h than that in ischemia/reperfusion group(P＜0.05).The expression level of Caspase-3 with the precondition of RSM was lower at 0h,3h,12h and 24h than that in NT group(P＜0.05).The level of ALT and AST with the precondition of RSM was higher than SO group at 3h,12h And 24h(P＜0.05) and was lower than the NT group at 3h,12h And 24h(P＜0.05).There was no change of organelles in SO group and N group.It has significant damage on organelles in NT group displayed by light microscope.And the damage in RSM group is slighter than the NT group.At 72h,the four groups have no special change except a few cell got light swell.ConclusionThe result showed that the expression of IGF-â… in RSM group was stronger than that in NT group.During hepatic ischemia reperfusion injury,the cellular damage was slighter and the hepatic function j better in RSM group than in NT group.The reason why may be RSM has a protective effect against hepatic ischemia-reperfusion injury by improve the microcirculation,improve the intracellular hypoxia environment, enhance the synthesis of IGF-â… in liver cell,up-regulate the expression of IGF-â… , inhibit cellular damage. |
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