Effects Of The Expression Of SOCS-3 In Erigeron Injection Applied To Rats With Focal Cerebral Ischemia-reperfusion Injury

Objective:Observe the dynamical expression and features of apoptotic cells and suppressor of cytokine signaling-3(SOCS-3) in the homolateral brain tissue after focal cerebral ischemia-reperfusion of rats; analyze the effects of Erigeron Injection on the expression of apoptotic cells and SOCS-3 and provide the basis for preventing and curing ischemia-reperfusion injury by Chinese crude drugs.Methods:Male SD rats (280-320g) were randomly divided into three groups:1) shame operation group (SG) 6 rats; 2) control group (CG) 42 rats, rats with simple right middle cerebral artery occlusion/reperfusion (MCAO/R); 3) experimental group (EG) 42 rats, apply Erigeron Injection to rats with right middle cerebral artery occlusion/reperfusion. One rat in every group was used to TTC staining, rats in CG and EG were subdivided into ischemic 2h reperfusion 3h,6h,12h,24h,72h,7d groups(n=6). The rats in EG were received Erigeron Injection 20MG/KG everyday two week before operation, then judge the neurological functional defect state in cerebral ischemia-reperfusion model made by modified thread embolism method when reperfusion. Referring to the longa[1] standard, rats who were scored 1-3 would be bring into experiment, but those would be rejected that scored 0 or 4, meet the score but there were SAH, cannot come around in 3h after the anaesthetic or die after operation. The neurological functional defects at 7 time points after reperfusion were assessed respectively. All these experimental rats were sacrificed in batch for obtaining their brain tissue, and then their brain tissue would be cut into four sections, one for the water content in brain measured by wet/dry weight, one for the apoptotic cells measured by TUNEL, one for the content of SOCS-3 measured by RT-PCR and one for pathological stain. The same method was used to deal with the right brain tissue of the rats in SG.Results:1)Praxiology score comparison:there was no neurological functional defect in SG, neurological functional defect exist both in CG and EG and their neurological functional scores in every time point were remarkably different (P<0.05).2) The water content in brain:the water content in the ipsilateral brain tissue of focus in CG and EG began to increase at 6h, and reached at their peak at 24h. But the degree of this increase was lower in EG compared with CG rats (P<0.05).3) Pathological changes:At 24h after operation, there was no pathological change took place in SG, while serious tissue destruction in ischemic region, intercellular space raritas, celluar swelling, nucleus minification, disappearance and inflammatory cells infiltration were found in CG, and in the EG rats, the pathologic features were presented much slighter than CG, intercellular space were found a litter broaden, and the degree of cell degeneration were presented much slighter compared with CG, including part of cells swelling and unclear nucleus and endochylema.4) The apoptosis mensuration of neurocyte:It was observed under light microscopy by TUNEL. The apoptotic cells were occasionally found in sham operation group, while the apoptotic cells in CG and EG began to increase gradually after reperfusion, and reached at their peak at 24h, then decrease gradually. By comparison, the distribution of apoptotic region and the number of apoptotic cells in EG is less than CG (P<0.05).5) The content change of SOCS-3mRNA:the expression of SOCS-3 was invariable in SG. In CG and EG, the expression of SOCS-3 increased significantly at the early stage, reached the peak at 24h, and then reduced gradually. But after 7d reperfusion, the expression was still higher than EG. Compared EG and CG, the expressive content of SOCS-3 in EG was higher than that in CG at every time point (P<0.05). Conclusions:1) Focal cerebral ischemia-reperfusion model made by modified thread embolism method is easy to operate and success, so it is the relatively ideal model to investigate cerebral ischemia-reperfusion injury.2) Rat cerebral ischemia-reperfusion can cause nerve functional lesion because of brain high-moisture capacity and apoptosis,3) Rat cerebral ischemia-reperfusion can cause high express of SOCS-3, which relieves Inflammatory reaction,brain oedema and apoptosis, neurone may be protected; 4)The Erigeron Injection could increase the expression of SOCS-3, because of the close relationship among anti-inflammation, edema, and inhibition of cell apoptosis.