| Objective:Inspect the dynamic variation of smad3and smad7in brain tissue after cerebral ischemia/reperfusion (I/R) in SD rats, analyze their correlation, explore their effects in cerebral I/R. Investigate the effects of Danhong injection on smad3and smad7after cerebral I/R in rats, clarify the possible mechanism of Danhong injection.Method:Randomly divide60250g-320g healthy SD rats into4groups:normal group (n=10), Sham-operated group (n=10), I/R group (n=20) and Danhong injection treatment group (n=20). Build the cerebral ischemia (2h) and reperfusion model in rats with Longa method. Further divide the I/R group and Danhong injection treatment group into4subgroups (5rats for each subgroup):6hours group (6h),1day group (Id),3days group (3d) and7days group (7d). Evaluate the nervus function coloboma of rats with the Bedersonl5-grades evaluation method. Investigate the pathology variation with HE staining method; Inspect the dynamic variation of the positive cells expression level of smad3and smad7with immunohistoch-emistry.The experimental results were analyzed with Spss17.0.Results:1. Danhong injection intervention group of rats neural function defect scale with time point obviously better than the ischemia reperfusion group. Danhong injection intervention group7days neural function defect scale is obviously better than the others each time point intervention group.(P<0.05)2. HE staining:normal group and Sham-operated group show that there is no obvious focus of infarct in the brain tissue, neuron structure and morphology are normal, no interstitial edema, I/R group shows neurons necrosis, and the lesions side neurons necrosis degeneration phenomenon is elevated and coupled with gliosis with time. And compared with subgroup with the same time point of I/R group, the treatment group shows obvious less brain tissue damage.3. Immunohistochemistry results:(1)Smad3occasionally shows positive cells in normal group and Sham-operated group. No difference between these two groups (P>0.05). Positive signal located mainly in the neurons and glia cytoplasm and the nuclei.The expression of smad3starts to increase in I/R6h group, reach the summit after3days, and then starts to decrease.7days group positive cells has the obvious decrease, but still higher than the normal group. Compared with the Sham-operated group and normal group, I/R group shows significantly (P<0.05) different smad3expression.(2) Smad7occasionally shows positive cells in normal group and Sham-operated group. No difference between these two groups (P>0.05). Positive signal located mainly in the neurons and glia cytoplasm and the nuclei. Smad7starts to increase rapidly in I/R6h group, reach the summit after3days, and then starts to decrease with time.7days group positive cells has the obvious decrease, but still higher than the normal group. Compared with the Sham-operated group, I/R group shows significantly (P<0.05) different smad7positive cell number.(3) In Dan-hong injection treatment group, the smad3expression time is compar-able with I/R group, but the positive cell number is significantly (P<0.05) higher than I/R group. And In Danhong injection treatment group and ischemia and reperfusion group Smad7expression of the different. In Danhong interention group h Smad76in the expression of rise somewhat, reach the summit after1days, and then starts to decrease with time.3days group and the7days group positive cells number significantly reduced, and below the IR group. The difference was statistically significant (P<0.01).Conclusions:1. Expressions of smad3and smad7both increase after cerebral I/R, smad3is a nerve to protect the role of the transformation growth factor beta1(TGF-β1) of the substrate, and combined TGF-β1with neural protection. And smad7is smads transcription inhibition, Can restrain smad3role. smad3will promote the expressions of smad7rise.2. Danhong injection treatment can make SD rats ischemia-reperfusion afterbrain organization smad3express rises, and smad7expression first increased and then decreased. |
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