Mechanism of Modified Si Junzitang on Neuronal Anti-anoikis of Cells in Rats with Cerebral Ischemia Through Fibulin-5 and p-Akt / 中国实验方剂学杂志

ABSTRACT

Objective:To investigate the effect of modified Si Junzitang on the expression of fibrous protein-5（Fibulin-5）， phosphorylated protein kinase B（p-Akt ）in hippocampus of rats after cerebral ischemia-reperfusion （I/R） injury and the anoikis of nerve cells. Method:The 60 male SD rats of SPF grade were randomly divided into sham operation group， model group， Edaravone group （3.2 mg·kg-1）and modified Si Junzitang high， medium and low-dose groups（19.08，9.54，4.77 g·kg-1）.The middle cerebral artery occlusion （MCAO） model was established by suture method，the rats were killed 7 days later，neurological deficit score was evaluated before the death，histopathological observation was performed by hematoxylin eosin staining， apoptosis index of nerve cells was detected by TdT-mediated dUTP nick end labeling（TUNEL）staining， the expression of Fibulin-5， p-Akt and protein in ischemic hippocampus were detected by immunohistochemistry and Western blot. Result:The neurological deficit score showed that，compared with the sham operation group， the neurological deficit score of the model group was significantly increased （P<0.01）， compared with model group， the neurological deficit score of Edaravone group，the high， medium， low dose groups of modified Si Junzitang were decreased （P<0.05，P<0.01）. Immunohistochemical results showed that，compared with the sham operation group， the expression of Fibulin-5， p-Akt protein and the apoptosis index of nerve cells in the model group were significantly increased （P<0.05，P<0.01）， compared with model group， the protein expressions of Fibulin-5 and p-Akt in Edaravone group， high， medium and low-dose groups of modified Si Junzitang were significantly increased （P<0.05，P<0.01）， and the apoptosis index of nerve cells was obvious，there was a significant decrease （P<0.05，P<0.01）. Western blot results showed that，compared with the sham operation group， the relative expression of Fibulin-5 and p-Akt protein in the model group was significantly down-regulated （P<0.01）， compared with model group， the protein expressions of Fibulin-5 and p-Akt in the Edaravone group， the high， medium and low-dose groups of modified Si Junzitang were significantly up-regulated （P<0.05，P<0.01）. Conclusion:The modified Si Junzitang may stabilize the extracellular matrix （ECM） Fibulin-5， increase the adhesion of ECM to cells and promote the expression of p-Akt protein， thus inhibiting neuronal apoptosis and protecting cerebral ischemia injury.