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zjandzq

金虫 (正式写手)

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In the present
study, we investigated the effect of Ap on the expression of the pi class of glutathione S-transferase (GSTP) in rat primary hepatocytes.
Hepatocytes were treated with 25 or 50 lg/mL of ethanol or ethyl acetate extracts of Ap (ApEE or ApEAE) or 10 or 20 lM androgra-
pholide, which is the major active diterpene lactone of Ap, for 48 h. ApEE, ApEAE, and andrographolide dose-dependently induced
GSTP protein and mRNA expression. In a GST activity assay, GST activity was significantly higher in cells treated with the maximum
concentrations of ApEE, ApEAE, and andrographolide than in control cells (P < 0.05). The pTA-2713 luciferase reporter construct con-
taining rat GSTP enhancer 1 (GPE1) was transiently transfected into Clone 9 liver cells. Cells treated with ApEE, ApEAE, and androg-
rapholide showed a dose-dependent increase in luciferase activity. GPE1 deletion abolished the induction efficiency of Ap. Also, the
induction of GSTP expression by Ap was inhibited by wortmannin, which is an inhibitor of the phosphatidylinositol 3-kinase
(PI3K)/Akt pathway. These results indicate that ApEE, ApEAE, and andrographolide induce GSTP expression. This induction is likely
related to the PI3K/Akt pathway, and GPE1, an enhancer element in GSTP promoter, is essential for the induction

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Miranda6276

木虫 (正式写手)

★ ★ ★ ★ ★ ★
zjandzq(金币+6,VIP+0):非常感谢! 6-15 21:48
在本研究中,我们考察了Ap对老鼠初级肝细胞中Pi类谷胱甘肽S转移酶表达情况的影响。肝细胞用25 or 50 lg/mL的Ap (ApEE or ApEAE)的乙醇或者是乙酸乙酯的萃取液处理48小时,或者用10 or 20 lM 的穿心莲内酯(Ap的唯一二萜内酯活性物质)处理48小时。ApEE, ApEAE和穿心莲内酯引起GSTP(谷胱甘肽S转移酶)和mRNA浓度依赖的表达。在GTS活性分析中,用最高浓度的ApEE, ApEAE和穿心莲内酯培养的 细胞中GTS的活性明显的比空白细胞中GTS活性高(P < 0.05). 含有鼠GSTP增强子的 pTA-2713 荧光素酶报告体系被转染到Clone 9肝细胞中。用ApEE, ApEAE和穿心莲内酯培养的细胞中荧光素酶表现出浓度依赖的活性增强现象。GPE1剔除使得Ap丧失了诱导活性。并且渥曼青霉素(一种磷脂酰肌糖激酶(PI3K)/Akt 路径抑制剂)可以抑制Ap对GSTP表达的诱导作用。这些结果说明ApEE, ApEAE和穿心莲内酯可以诱导GSTP表达,这种诱导过程可能与磷脂酰肌糖激酶(PI3K)/Akt 路径有关,并且GPE19(一种GSTP促进因子的增强子)对该诱导过程十分重要。
2楼2009-06-15 12:03:56
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