Study Of Expressions Of HSP10, Caspase-8 And DR5 Proteins And The Effects Of Dl-3-n-butylphthalide In Brain Tissue Following Cerebral Ischemic Reperfusion In Rats

Objective:To investigate the expression of the heat shock protein 10(HSP10), Caspase-8 and DR5 proteins in the rat brain tissues following global cerebral ischemic reperfusion and treatment of dl-3-n-butylphthalide, and to study the molecular mechanism of the curative effects of dl-3-n-butylphthalide.Methods:(1) Animal Groups:Fifty healthy SD rats were divided into four groups:Normal Group;Sham Operation Group; Ischemia Reperfusion Group (I/R); NBP Intervention Group (NI/R); Further, the I/R Groupand NI/R Group are divided into four subgroups:six hours, one day, three days and seven days after I/R injury.(2) Models:15 minutes'common carotid artery occlusion(CAO) of both sides and reperfusion for six hours, one day, three days and seven days. Sodium Nitroprusside(SNP) control low blood pressure in rats. The sham operation group only give segregation but no occlusion; gavage of NBP (80mg/kg, BID) until death.(3) H&E staining and immunohistochemical detection of NSE expression in neurons were used to count the numbers of neurons and characteristic of tissue injury in each group under the light microscope.(4) The expressions of Caspase-8,DR5 and HSP10 proteins in brain tissue were detected by immunohistochemistry technique. Then the quantitative analysis was performed by Image Analysis System.(5) Two-sided T test was examined all data using statistical software package SPSS 12.0, P≤0.05 was considered statistically signification. Results:(1) Significant glial cells hyperplasia, interstitial edema, cellular edema and manipulus focal necrosis were observed in the I/R group by increasing ischemic times. But, on the same time point, there were much mild degree of neuronal degeneration, glial cell proliferation and brain tissue necrosis in the each subgroups of NBP intervention compared with I/R Group (P<0.05)(2) There was low expressions of HSP10 protein in both normal control group and sham operation group, also, there was no significant difference between normal control group and sham operation group (P>0.05). Increasing expression of HSP10 was conformed in I/R group compared with the sham operation group in the rat brain tissues by treatment of different time points, such as level of proteins expression was increasing in the six hours, the expression level reached the peak in the third day and kept the high expression level until seven days. Also, there was significant difference between I/R group and the sham operation group (P<0.05).(3) There were low expressions of Caspase-8 and DR5 proteins in both normal control group and sham operation group, also, there was no significant difference between normal control group and sham operation group (P>0.05). Increasing expression of Caspase-8 and DR5 were conformed in I/R group compared with the sham operation group in the rat brain tissues by treatment of different time points, such as level of proteins expression was increasing in the six hours, the expression level reached the peak in the third day, but the expression level was decreasing from third day until seventh day. Also, there was significant difference between I/R group and the sham operation group (P<0.05).(4) The increasing trendy of expression HSP10 protein in the NBP group was somewhat alike to I/R group, but, on the same treatment of time points, the positive expression of HSP10 was significantly high in NBP group than that of in I/R group (P<0.05). The decreasing trendy of expression Caspase-8 and DR5 in the NBP group were similar with I/R group, but, on the same treatment of time points, the positive expression of Caspase-8 and DR5 were significantly low in NBP group than that of in I/R group (P<0.05).Conclusions:1. After Treatment of Ischemia-Reperfusion was experimented in the rats, the expression of HSP10, Caspase-8 and DR5 proteins were evidently increase in the brain tissue of experimental groups compared with control groups.2. The curative effects of NBP in rat's cerebral ischemia-reperfusion injury might be increase expression of HSP10 protein and decrease the expression of Caspase-8 and DR5 proteins to inhibit cell apoptosis and reduce neuronal death.