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HSYA A on endothelial cells after hypoxia and hypoxia inducible factor expression and regulation mechanism of

Author: LianZeQin
Tutor: ZhuHaiBo；ZhangLing；ZhaoDaLong
School: Peking Union Medical College , China
Course: Pharmacology
Keywords: Hydroxysafflor yellow A hypoxia-inducible factor 1α von Hippel-Lindau gene tumor suppressor gene p53 phosphatidylinositol 3 kinase protein kinase B phosphatase and tensin homolog deleted on chromosome ten Eahy 926 cell line
CLC: R285.5
Type: Master's thesis
Year: 2008
Downloads: 102
Quote: 0
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Abstract


AIM.Hydroxysaffior yellow A is one of the main active ingredients extracted from Carthamus tinctorius.L.Our preliminary work suggested that it exerted a good protective effect on cerebral ischemia in rats by enhancement on angiogenesis around cerebral ischemic infarct area.In present study we investigated the mechanism involved in Hydroxysafflor yellow A（HSYA） regulating HIF-1αexpression in EAhy 926 cell line under 1%O2 hypoxia.Methord:Eahy 926 cells were incubated under hypoxia for different time after treated with HSYA(100μmol·L-1,10μmol·L-1,1μmol·L-1 ).Cell growth rate was detected using MTT assays.VHL,p53,PI3K,Akt and PTEN location and protein expression were analyzed by Immunocytochemical stain.RT-PCR was performed to investigate HIF-1α, VHL,p53,PI3K,Akt and PTEN mRNA expression.Protein expression of HIF-1α,VHL, p53,PI3K,Akt and PTEN were evaluated by Western-blotting method.Distribution of HSYA in culture EAhy926 cells was detective by HPLC.Metabolite profiling of MCAO rat cortex after treated with HSYA using high was collect using high resolution NMR combined with multivariate statistical methods.Result:HSYA at the concentration of 100μmol·L-1 promoted Eahy 926 cells growth rate under hypoxia.HIF-1αmRNA and protein expression was up-regulated in the presence of HSYA.HSYA shows down-regulation effect on VHL,p53 and PTEN and up-regulation on PI3K and Akt.Time series research showed that HSYA present strong relevance with hypoxia duration.Intracellular HSYA concentration was under the limit of quantification.HSYA could up-regulated lactic acid level and down-regulated glucose of cortex after ischemic in MCAO rat.Discussion:HSYA exerted an effect on protecting Eahy 926 cell from hypoxia as well as its up-regulating on HIF-1αexpression might be partially associated with its inhibition on VHL,p53 and PTEN expression and promotion onPI3K,Akt expression.As a water-soluble molecular,HSYA showed a lack of permeability through membrane.The metabonomic approach based on NMR profiling of MCAO rat cortex samples represents a powerful up-regulated on glucose and down-regulated on lactic acid,providing relevant information on the effects of HSYA on endothelia protection and HIF-1αregulation.

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