The Regulatory Mechanism Study Of Hydroxysafflor Yellow A On Inflammation Pathway Related Factors In Permanent MCAO Rats

Hvdroa:vsafflor yellow A(HSYA),is a mono-active component of flower,Carthamus tinctorius L.Recent years,the pharmacology mechanism of HSYA has been widely studied,especially on its anti-cerebral ischemia effect.The mechanisms involved in this anti-cerebral ischemia neuroprotection effect contain its anti-platelet aggregation,anti-oxidative damage as well as anti-excitatory amino acids toxicity and so on.However,the exactly mechanism involved was still unknown and there's less the study on its anti-inflammation after cerebral ischemia.So with the suggestion of our previous study on microarray after cerebral occlusion that HSYA may regulate the post ischemic inflammatory cytokines to achieve its neuroprotection,we use a rat permanent focal cerebral ischemia model which using intraluminal suture occlusion method to examine its anti-inflammation action.Firstly,we compared the different ways of HSYA administration and chose the successive administration to study HSYA's mechanism which was the most effective way.After cerebral artery occlusion 3h,6h,12h and 24h, cortex was removed for the next experiments.Western blotting was used to detect the expression of p65 protein and the phospho-IκB-α(pIκB-α) in the cytoplasm or the nucleus.NF-κB DNA binding activity was measured by Trans-AM transcription factor assay kits.mRNA and protein expression of cytokines TNF-α,IL-1β,IL-6 and IL-10 was measured by the RT-PCR and ELISA method.Moreover,we compared HSYA with the NF-κB inhibitor Pyrrolidine dithiocarbamate ammonium salt(PDTC) on the inhibition effect.Finally,HPLC was used to determine the contents of HSYA in plasma and in brain after rat MCAO and NMR was used to detect the metabolic substances.The result showed that successive intravenous injection of HSYA(10mg·kg^-1) to rats after cerebral occlusion,the p65 translocation activity and the phosphorylation of IκB-αwere significantly inhibited.At the same time,HSYA suppressed p65 binding activity and the transcriptional and protein level of pro-inflammatory cytokines:TNF-α,IL-1β,IL-6 and promoted both the mRNA and the protein expression of anti-inflammatory cytokine IL-10.HSYA also showed an effect of inhibiting the toxicity metabolic substances lactic acid and glutamic acid but elevate the GABA and glucose.In conclusion,our study suggested that the inhibition of NF-κB activity and the mRNA and protein expression of cytokines in the inflammatory transduction pathway as well as inhibition of the production of toxicity substances to maitein the energy metabolism of the ischemic brain probably explained the anti-cerebral ischemic mechanism of HSYA.