AIM:To assess effects of heme on messenger RNA(mRNA) and microRNA(miRNA) profiles of liver cells derived from humans.METHODS:We exposed human hepatoma cell line Huh-7 cells to excess iron protoporphyrin(heme)(10 μmol...AIM:To assess effects of heme on messenger RNA(mRNA) and microRNA(miRNA) profiles of liver cells derived from humans.METHODS:We exposed human hepatoma cell line Huh-7 cells to excess iron protoporphyrin(heme)(10 μmol/L) or induced heme deficiency by addition of 4,6-dioxoheptanoic acid(500 μmol/L),a potent inhibitor of aminolevulinic acid dehydratase,for 6 h or 24 h.We harvested total RNA from the cells and performed both mRNA and miRNA array analyses,with use of Affymetrix chips,reagents,and instruments(human genome U133 plus 2.0 and miRNA 2.0 arrays).We assessed changes and their significance and interrelationships with Target Scan,Pathway Studios,and Ingenuity software.RESULTS:Changes in mRNA levels were most numerous and striking at 6 h after heme treatment but were similar and still numerous at 24 h.After 6 h of heme exposure,the increase in heme oxygenase 1 gene expression was 60-fold by mRNA and 88-fold by quantitative reverse transcription-polymerase chain reaction.We found striking changes,especially up-regulation by heme of nuclear erythroid-2 related factor-mediated oxidative stress responses,protein ubiquitination,glucocorticoid signaling,P53 signaling,and changes in RNAs that regulate intermediary metabolism.Fewer mRNAs were down-regulated by heme,and the fold decreases were less exuberant than were the increases.Notable decreases after 24 h of heme exposure were patatin-like phospholipase domain-containing protein 3(-6.5-fold),neuronal PAS domain protein 2(-1.93-fold),and protoporphyrinogen oxidase(-1.7-fold).CONCLUSION:Heme excess exhibits several toxic effects on liver and kidney,which deserve study in humans and in animal models of the human porphyrias or other disorders.展开更多
INTRODUCTIONLiver fibrosis is a dynamic course leading tocirrhosis from a various chronic liver diseases. Thepathological basis of fibrosis is the disturbance ofproduction and degradation of the extracellularmatrix (E...INTRODUCTIONLiver fibrosis is a dynamic course leading tocirrhosis from a various chronic liver diseases. Thepathological basis of fibrosis is the disturbance ofproduction and degradation of the extracellularmatrix (ECM), which causes accumulation of ECMin the liver[1,2].展开更多
目的:观察中药复方更年健对老年雌性大鼠下丘脑、垂体和卵巢雌激受体(ER)和 ER mRNA 的影响。方法:建立自然衰老的雌性大鼠模型,用放射配体结合分析法检测 ER,用 Norther blot 分析 ER mRNA 的改变。结果:雌性大鼠性减退期下丘脑,垂体...目的:观察中药复方更年健对老年雌性大鼠下丘脑、垂体和卵巢雌激受体(ER)和 ER mRNA 的影响。方法:建立自然衰老的雌性大鼠模型,用放射配体结合分析法检测 ER,用 Norther blot 分析 ER mRNA 的改变。结果:雌性大鼠性减退期下丘脑,垂体和卵巢ER 和 ER mRNA 水平随血清雌二醇(E_2)水平的下降较性成熟期显著下降,而更年健则使下降了的 ER 和 ER mRNA 水平明显提高。结论:本结果提示更年健在临床上治疗更年期综合征的机理可能是在不改变 E_2水平的情况下,提高下丘脑、垂体和卵巢 ER 和 ER mRNA 水平,从而增强雌激素的生物学效应,改善临床症状和体征。展开更多
基金Supported by A Grant from NIH/NIDDK (DK38825) to Bonkovsky HLInstitutional Funds from the Carolinas Health Care Foundation and Carolinas Medical Center
文摘AIM:To assess effects of heme on messenger RNA(mRNA) and microRNA(miRNA) profiles of liver cells derived from humans.METHODS:We exposed human hepatoma cell line Huh-7 cells to excess iron protoporphyrin(heme)(10 μmol/L) or induced heme deficiency by addition of 4,6-dioxoheptanoic acid(500 μmol/L),a potent inhibitor of aminolevulinic acid dehydratase,for 6 h or 24 h.We harvested total RNA from the cells and performed both mRNA and miRNA array analyses,with use of Affymetrix chips,reagents,and instruments(human genome U133 plus 2.0 and miRNA 2.0 arrays).We assessed changes and their significance and interrelationships with Target Scan,Pathway Studios,and Ingenuity software.RESULTS:Changes in mRNA levels were most numerous and striking at 6 h after heme treatment but were similar and still numerous at 24 h.After 6 h of heme exposure,the increase in heme oxygenase 1 gene expression was 60-fold by mRNA and 88-fold by quantitative reverse transcription-polymerase chain reaction.We found striking changes,especially up-regulation by heme of nuclear erythroid-2 related factor-mediated oxidative stress responses,protein ubiquitination,glucocorticoid signaling,P53 signaling,and changes in RNAs that regulate intermediary metabolism.Fewer mRNAs were down-regulated by heme,and the fold decreases were less exuberant than were the increases.Notable decreases after 24 h of heme exposure were patatin-like phospholipase domain-containing protein 3(-6.5-fold),neuronal PAS domain protein 2(-1.93-fold),and protoporphyrinogen oxidase(-1.7-fold).CONCLUSION:Heme excess exhibits several toxic effects on liver and kidney,which deserve study in humans and in animal models of the human porphyrias or other disorders.
基金Project supported by the National Natural Science Foundation of China, No. 39500138
文摘INTRODUCTIONLiver fibrosis is a dynamic course leading tocirrhosis from a various chronic liver diseases. Thepathological basis of fibrosis is the disturbance ofproduction and degradation of the extracellularmatrix (ECM), which causes accumulation of ECMin the liver[1,2].
文摘目的:观察中药复方更年健对老年雌性大鼠下丘脑、垂体和卵巢雌激受体(ER)和 ER mRNA 的影响。方法:建立自然衰老的雌性大鼠模型,用放射配体结合分析法检测 ER,用 Norther blot 分析 ER mRNA 的改变。结果:雌性大鼠性减退期下丘脑,垂体和卵巢ER 和 ER mRNA 水平随血清雌二醇(E_2)水平的下降较性成熟期显著下降,而更年健则使下降了的 ER 和 ER mRNA 水平明显提高。结论:本结果提示更年健在临床上治疗更年期综合征的机理可能是在不改变 E_2水平的情况下,提高下丘脑、垂体和卵巢 ER 和 ER mRNA 水平,从而增强雌激素的生物学效应,改善临床症状和体征。
