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OALib Journal期刊
ISSN: 2333-9721
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三七总皂甙增强人内皮源性一氧化氮合酶基因启动子活性
, PP. 1113-1116
Keywords: 三七总皂甙,一氧化氮合酶,基因表达调控,酶学,共转染
Abstract:
目的探讨三七总皂甙(tpns)对人内皮源性一氧化氮合酶(enos)基因启动子转录活性的调控机制。方法以基因重组技术构建人enos基因启动子区(-1~-1600bp)驱动的萤火虫荧光素酶报告基因载体penos-luc,采用脂质体介导的细胞基因共转染技术将penos-luc、空载体pgl2-basic和β-半乳糖苷酶表达质粒pcmv-β共转染nih3t3细胞,用细菌脂多糖(lps)、tpns和转移生长因子β1(tgfβ1)等因子分别刺激转染后的nih3t3细胞,检测并比较荧光素酶/β-半乳糖苷酶活性,以确定lps、tpns和tgfβ1对人enos基因启动子转录活性的影响。结果(1)酶切和测序结果均证实重组载体penos-luc构建正确;(2)重组载体penos-luc在nih3t3细胞中有效表达;(3)在lps刺激下,人enos启动子的转录活性降低,而在tpns和tgfβ1刺激下,其启动子的转录活性增强,其中,tgfβ1较tpns所致的转录活性更强。结论正确构建了人enos基因启动子区(-1~-1600bp)驱动的萤火虫荧光素酶报告基因载体penos-luc;tpns在nih3t3细胞中增强人enos基因启动子的转录活性。
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