Zhu Dan-Yan, Du Yue, Huang Xin, Guo Mei-Yuan, Ma Kui-Fen, Yu Yong-Ping, Lou Yi-Jia
Institute of Pharmacology & Toxicology and Biochemical Pharmaceutics, Zhejiang University, Hangzhou, China.
Stem Cells Dev. 2008 Aug;17(4):775-83. doi: 10.1089/scd.2007.0241.
The expressions of membrane-associated proteins in eicosanoid and glutathione metabolism (MAPEG), a superfamily involved in both inflammation and cell protection, were investigated in an in vitro system of mouse embryonic stem (ES) cell-derived hepatic tissue. Gene expressions of all MAPEG members were demonstrated in a developmental-dependent manner in the derived hepatic tissue. The protein expression of microsomal glutathione S-transferase 1 (MGST1) was not detected until differentiating day 14. It gradually increased by maturation of hepatic tissue. The microsomes of ES cell-derived hepatic tissue possessed the MGST1-like catalytic activity. However, MGST1 from the microsome preparation could not form dimers as usual when exposed to reactive nitrogen species ONOO. Among the other members in MAPEG, weak expressions of leukotriene C(4) synthase (LTC(4)S) and microsomal prostaglandin E synthase 1 (mPGES-1) were observed. A stable expression of 5-Lipoxygenase activating protein (FLAP) appeared during the entire course of differentiation. MGST2 and MGST3 failed to express in the derived hepatic tissue, although mRNA of them do existed. In conclusion, ES cell-derived hepatic tissue possess MAPEG gene expression features, but not all protein expression could be detected, which helps to understand not only the nature of the tissue derived, but also the fate of bioartificial liver system, and may as well provide a valuable in vitro model for research in both inflammation process and toxic events in hepatological fields.
在小鼠胚胎干细胞衍生肝组织的体外系统中,研究了类花生酸和谷胱甘肽代谢相关膜蛋白(MAPEG)的表达,MAPEG是一个参与炎症和细胞保护的超家族。在衍生的肝组织中,所有MAPEG成员的基因表达均呈发育依赖性。直到分化第14天,微粒体谷胱甘肽S-转移酶1(MGST1)的蛋白表达才被检测到。随着肝组织成熟,其表达逐渐增加。胚胎干细胞衍生肝组织的微粒体具有MGST1样催化活性。然而,当暴露于活性氮物质ONOO时,微粒体制备物中的MGST1不能像往常一样形成二聚体。在MAPEG的其他成员中,观察到白三烯C4合酶(LTC4S)和微粒体前列腺素E合酶1(mPGES-1)的弱表达。在整个分化过程中,5-脂氧合酶激活蛋白(FLAP)呈现稳定表达。尽管MGST2和MGST3的mRNA存在,但在衍生的肝组织中未能表达。总之,胚胎干细胞衍生的肝组织具有MAPEG基因表达特征,但并非所有蛋白表达都能被检测到,这不仅有助于了解所衍生组织的性质,还有助于了解生物人工肝系统的命运,并且可能为肝脏领域炎症过程和毒性事件的研究提供有价值的体外模型。