| Objective : By establishing the model of subarachnoid hemorrhage(SAH) in rats to detect the dynamic change of PICP concentration in cerebrospinal fluid(CSF) and the expression of Wnt3a、β- catenin protein in the brain tissue,and to explore the role of Wnt /β- catenin signaling pathway in leptomeningeal fibrosis in rats after suba rachnoid hemorrhage,which may provide more theoretical basis to treat chronic hydrocephalus after SAH through the anti-fibrosis treatment way.Methods : 1.To establish the subarachniod hemorrhage model of rats by double autologous blood injection into cisterna magna.2.The 75 SD healthy male rats were randomly divided into three groups:blank group,sham operation group,model group.the three groups were distributed 25 rats each, observed at five time points, which were 3d、6d、10d 、 14 d and 21 d after operation.In the corresponding time points after operation,cerebrospinal fluid of each rat was collected for testing,then Fixed and removed brain tissue,paraffin embedded and section.3.The succession of SAH model was estimated by observing the general conditions and performing the behavioral scores of rats in each group at 1d,3d,6d,10 d,14d,21 d after operation.4.Masson staining was performed to the twenty-first day of the rat brain slices of each group,including blank group,sham operation group and model group,we analysised the average gray and thickness of each group leptomeningeal collagen fiber by using Motic image advances 3.2 imageanalysis system, and carried on the quantitative analysis.5.Using radioimmunoassay to measure the levels of PICP in collected CSF of blank group,sham operation group, model group at each time point.6.Using immunohistochemical staining to observed the expression of wnt3 a,β-catenin protein in brain tissue of blank group,sham operation group and model group at each time point.Rerults:1.The rats of model group were listlessness,less activity and poor appetite.at 1d,3d,the behavioral scores of model group was significantly lower than that of sham operation group and blank group(P<0.01),and at 6d,10 d,14d,21 d,there was no statistical significance between each group(P>0.05).2.Masson staining of the twenty-first day rat leptomeningeal: The average gray of model group leptomeningeal was significantly lower than that of sham operation group and blank group, the average thickness was significantly higher than that in sham operation groupand blank group(P<0.01);The average content of collagen fiber of leptomeningeal in model group was significantly higher than that in sham operation group and blank group(P<0.01).The average gray,average thickness and average collagen fiber content had no statistical significance between the sham operation group and the blank group(P>0.05).3.The change of PICP concentration:The PICP concentration of model group at each time point in CSF was significantly higher than that in sham operation group and blank group(P<0.01).There was no statistically significant difference between the sham operation group and the blank control group(P>0.05).4.The expression of Wnt3 a protein: The expression of Wnt3 a protein of model group significantly increased at each time point compared with that in sham operation group and blank group(P<0.01),There was a positive correlation between Wnt3 a protein and PICP in model group( P<0.01).The sham operation group and the blank control group have no statistically significant difference(P>0.05).5.The expression of β-catenin protein: The expression of β-catenin protein of model group significantly increased at each time point compared with that in sham operation group and blank group(P<0.01),There was a positive correlation between Wnt3 a protein、β-catenin protein and PICP in model group( P<0.01). Between the sham operation group and the blank control group,the difference was not statistically significant.(P>0.05).Conclusion:1.The SAH model of rat established by double autologous blood injection into cisterna magna is very simple and operational, the pathophysiology in this model is nearly as same as human, thus it can be used generally.2.The levels of PICP in CSF elevated after SAH,which prompted the fibrosis of leptomeningeal. There was a correlation between the Continuous high expression of PICP and the severity of fibrosis.The continuous occurrence and accumulation of leptomeninges fibrosis may accelerate chronic hydrocephalus after the formation of SAH.3.There was a positive correlation between the increase of Wnt3 a protein、β-catenin protein and PICP in model group,which prompt that the activation of Wnt3a/ β-catenin signaling pathway might play an important role in the development of leptomeningeal fibrosis after SAH. |
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