The Relation Between The Bleeding Amount Of Experimental Intracerebral Hemorrhage In Rats And Nse, S100 As Well As The Effect Of Tcm Intervention

Purpose:On the basis of Eighth Five Year's Plan",this research aims to investigate the evidence and mechanism of Chinese internal medicine coNSErvative treatment on ICH within critical amount of bleeding(<50ml);Research the change of NSE, serum S100, protein expression and regulation of gene expression before and after the TCM intervention and the correlation with the bleeding amount and NDS,which to confirm the optimum critical point of the sensibility and the specific change of serum when ICH causes damage on Brain Parenchyma, and to further probe into the correlation between the Traditional Chinese medical treatment and the optimum ICH critical amount of bleeding, and the diagnostic criteria in specificity and sensibility.Method:(1) A stable and feasible ICH rats model is built to observe the NIHSS and brain tissue pathomorphology change before and after the intervention of Traditional Chinese Medicine. (2) By means of ELIS A to detect NSE and S100 serum values; By means of SP to detect the protein expression n brain tissue of NSE and S100 in brain tissue of positive expression; By means of RT-PCR to detect a dynamic change of mRNA regulation in brain tissue NSE,S100. (3)By comparing with the preliminary experiment, the amount of bleeding, the time of bleeding and the dynamic change can be observed and the critical point of serum change when the brain parenchyma is damaged, and the optimum amount of bleeding and the best diagnostic criteria with the treatment of Traditional Chinese Medicine can be confirmed.Results:(1) Five days after the modeling,the model group,there is significant difference(p<0.01) in NIHSS between the model group and the treatment group with sham group. When the amount of bleeding are the same,the model group's NIHSS is higher than the treatment group and the difference is significant(P<0.05).With the gradual increasing of the amount of bleeding,NIHSS of model group and treatment group both have the trend of increasing. (2) The microscopic HE stain reveals that the brain tissues in the model group and the treatment group have been damaged when comparing with the sham group,but the level of the brain damage in treatment group is far less than that in model group.(3)Five days after the modeling,the serum NSE levels of model group and treatment group are higher than that of sham group.and the difference is significant (p<0.01).With the increase of bleeding amount, the serum level of the former two groups also increases. A comparison has been made inside the model group and there is significant difference (P<0.01) in the amount of bleeding between 40μl.53μl.67μl and 27μl, while there is no significant difference (P>0.05) betweer 40μl.53μl and 67μl.After using Traditional Chinese Medicine.when compared with the model group.the serum NSE level of the treatment group has been decreased;and in comparison with the different bleeding amount,there is significant difference (p<0.01) between model group and treatment group in 27μl> 40μl.When the bleeding amount is 53μl,there is significant difference(P<0.05)between model group and treatment group,and when the bleeding amount is 67μl,there is no statistic point between model group and treatment group.The result of immunohistochemistry shows that except 27μl in treatment group,the rats' brain tissue NSE optical density in other groups are higher than sham group.and NSE is increased;the more the bleeding amount is,the stronger the NSE;after using the Traditional Chinese Medicine.the NSE in treatment group is weaker than that of model group.when compared between model group and treatment group.the bleeding amount with 27μl>40μl is more significant(P<0.01). NSE semiquantitative PCR result indicates that five days after ICH,the levels of transcription of NSE in model groups increase significantly,with the increase of bleeding amount.it gradually increases.When the bleeding amount is the same,compared with the corresponding model group,NSE transcriptional level of TCM treatment group has been in decrease. (4)Five days after modeling.the serum S100 in model group and treatment group is higher than the sham group;with the increase of bleeding amount.the level of the serum S100 in model group and treatment group gradually increases.With the intervention of TCM, the level of serum S100 decreases,especially significant (P<0.01) in less bleeding amount (27μl,40μl).The result of SP shows that the rats'brain tissue S100 in treatment group and model group is higher than sham group. With the increase of bleeding amount.S100 is stronger;after TCM treatment 100 in treatment group decreased relatively compared with model group;S100 semiquantitative PCR result indicates that five days after ICH.S100 transcriptional level in model groups is stronger compared with sham group. With the increase of bleeding amount,it gradually increases. When the bleeding amount is the same, compared with the corresponding model group.S 100 transcriptional level of TCM treatment group has been in decrease. (5) Compared between NSE and S100 density level three days and five days after rats' ICH in model group,it is found that when the rats' bleeding amount is 27μl,the serum density level is lower than the other groups(P<0.01或P<0.05),when the bleeding amount reaches 40μl,the serum density level has been leveled off.And there is no statistic point in the bleeding amount between 40μL 53μl, and 67μl.There is significant difference (p<0.01) in rats'serum density level between treatment group and model group when the bleeding amount is 27μl and 40μl before and after TCM treatment.There is significant difference (p<0.05) or no significant difference (p>0.05) when the bleeding amount is 53μl and 67μl.When the ROC curve analysis is made in serum NSE and S100 density level,serum NSE density level ranges from 19.95ng/ml to 21.65ng/ml,and the serum S100 density level ranges from 6.3ng/ml to 8.16ng/ml.Conclusion:(1) It preliminarily deduced that 40μl in ICH rats (i.e.30ml in human ICH) is the critical point of specificity and sensitivity of NSE and S100 when the brain parenchyma has been damaged, which provides the valuable reference data for clinical diagnosis. (2) It preliminarily established that the optimum sensitive point of Detection of NSE and S100 is three days after ICH. which provides the best treatment for clinical diagnosis of opportunity. (3) It preliminarily deduced that less than 40μl in ICH rats (i.e.30ml in human ICH) is the best curative effect with TCM on ICH, which provides the basis for clinical choose reasonable and effective treatment methods. (4) It preliminarily deduced that when the serum NSE density level is between 19.95ng/ml-21.65ng/ml and the serum S100 density level is between 6.3ng/ml-8.16ng/ml, that is the safe and effective interval with TCM on ICH and which provides the objective diagnostic criteria for clinical treatments.