Study On Effects Of PAMD On Apoptosis And Expression Of Its Relative Proteins And Genes In Experimental Rats With MCAO

Objective: To study and explore the neuroprotective effects and the further mechanism of phenolic alkaloids from Menispermum dauricum (PAMD) on cerebral ischemia in rats.Methods: In order to observe the neuroprotective effect of PAMD on cerebral ischemia, the rats with incomplete cerebral ischemia were induced with fishing lines in the middle cerebral artery (MCA) and then were executed at 24h, 48h and 72h after cerebral ischemia. It was observed the effect of PAMD on the changes of ethology and pathomorphology due to nerve injury of rats with MCAO before and after treatments. HE staining was used to observe the changes of histomorphology. Electronic microscopy was used to study the ultrastructure of areas. It was detected the effects of PAMD on protein expression of heat shock protein 70 (HSP70), protein kinase B (PKB) ,c-fos and p53 with immunohistochemical method. In addition, we observe the rate of cell apoptosis with flow cytometry and the gene expression of Bcl-2 mRNA and Bax mRNA with reverse transcription polymerase chain reaction (RT-PCR) at 24h after cerebral ischemia.Results: PAMD could make obviously pathomorphology of cerebral ischemia better, improve change of ethology, decrease the rate of cell apoptosis of ischemic cerebral tissue, inhibit the expression of c-fos,p53 protein and increase the expression of HSP70,PKB protein in cerebral cortex. And PAMD can efficiently increase expression of Bcl-2 mRNA and decrease the expression of Bax mRNA in cortical neuron to protect the nerves after cerebral ischemic injury of rats.Conclusion: PAMD had protecting effect on cerebral ischemia and the mechanism of PAMD against cerebral ischemia is related to inhibiting cell apoptosis and affecting the related expression of gene and protein interrelated with cell apoptosis.