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国家自然科学基金(30370676)

作品数:4 被引量:8H指数:2
相关作者:韩晓杨晓敏林玲吕京澴更多>>
相关机构:南京医科大学更多>>
发文基金:国家自然科学基金国家重点基础研究发展计划更多>>
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Nono蛋白的原核表达、纯化和多克隆抗体的制备被引量:2
2007年
目的表达和纯化带多聚组氨酸(6×His)标签的Nono(non-POU-domain-containing,octamer-bindingprotein)融合蛋白并制备抗Nono多克隆抗体。方法构建pET-28a(+)-Nono重组表达质粒,转入Rosetta(DE3)大肠埃希菌,以IPTG诱导6×His-Nono融合蛋白表达,经镍离子金属螯合树脂纯化后,用纯化出的蛋白免疫BALB/C小鼠制备多克隆抗体,并用ELISA检测多克隆抗体的效价,Western印迹检测多克隆抗体的特异性。结果在大肠埃希菌中诱导出高水平表达的His-Nono融合蛋白,经亲和树脂纯化后免疫小鼠,获得了高特异性的抗Nono抗血清。结论成功构建pET-28a(+)-Nono原核表达质粒,表达并纯化出高纯度的目标蛋白,制备出高滴度、高特异性的多克隆抗体。
林玲杨晓敏吕京澴韩晓
关键词:COX-2基因基因重组融合蛋白多克隆抗体
应激状态下环加氧酶-2基因表达调控的研究进展被引量:3
2005年
环加氧酶-2是体内催化花生四烯酸合成前列腺素的关键酶。它在细胞因子、有丝分裂原、生长因子等诱导刺激下可以大量表达,并在炎症和肿瘤的发生发展过程中发挥着重要作用。近年来的研究表明,应激状态下,环加氧酶-2转录增强和(或)mRNA稳定性增强,导致其蛋白表达量增加及产物前列腺素增多,从而引起机体一系列的生理和病理反应。
杨晓敏韩晓
关键词:环加氧酶-2应激状态基因表达调控转录因子
Resveratrol prevents interleukin-1 β-induced dysfunction of pancreatic β-cells被引量:3
2010年
Objective:Interleukin-1β(IL-1β)plays an important role in the development of type 1 and type 2 diabetes mellitus.Resveratrol,a polyphenol,is known to have a wide range of pharmacological properties in vitro.In this research,we examined the effects of resveratrol on IL-1β-inducedβ-cell dysfunction.Methods:We first evaluated the effect of resveratrol on nitric oxide(NO)formation in RINm5F cells stimulated with IL-1βusing the Griess method.Next,we performed transient transfection and reporter assays to measure the transcriptional activity of peroxisome proliferator-activated receptor-γ(PPAR-γ).We also used Western blotting analysis to assess the effect of resveratrol on inducible nitric oxide synthase(iNOS)expression and nuclear factor-κB(NF-κB)translocation to the nuclei in cells treated with IL-1β.In addition,we assessed the transcriptional activity of NF-κB using an electrophoretic mobility shift assay(EMSA).Finally,we evaluated the effect of resveratrol on IL-1β-induced inhibition of glucose-stimulated insulin secretion in freshly isolated rat pancreatic islets.Results:Resveratrol significantly suppressed IL-1β-induced NO production,a finding that correlated well with reduced levels of iNOS mRNA and protein.The molecular mechanism by which resveratrol inhibited iNOS gene expression appeared to involve increased PPAR-γactivity,which resulted in the inhibition of NF-κB activation.Further analysis showed that resveratrol could prevent IL-1β-induced inhibition of glucose-stimulated insulin secretion in rat islets.Conclusion:In this study,we demonstrated that resveratrol could protect against pancreaticβ-cell dysfunction caused by IL-1β.
Fang ChenXiaohua ZhouYan LinChangwen JingTao YangYong JiYujie SunXiao Han
关键词:过氧化物酶体增殖物激活受体一氧化氮合成酶
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