全反式视黄酸对大鼠原代肝细胞铁代谢的影响

Effect of all-trans retinoic acid deficiency on iron metabolism in rat primary hepatocyte

目的研究全反式视黄酸(atRA)对大鼠原代肝细胞铁代谢的影响。方法按seglent胶原酶消化法分离大鼠原代肝细胞,将活性大于90%的肝细胞培养于10%血清DMEM培养基中,随机分为4个剂量组,即维生素A完全缺乏组、边缘性缺乏组、正常对照组、治疗剂量组,分别给予0、0.5、1.0和50μmol/L的atRA。收集培养72h后的肝细胞,用RT-PCR法检测铁调节蛋白2(IRP2)mRNA、转铁蛋白受体(TFR)mRNA、铁蛋白(Fn)mRNA的表达。结果VA缺乏时可导致肝细胞的活性及合成功能降低,使IRP2 mRNA、TFR mRNA表达增加,而Fn mRNA表达下降;而补充atRA可抑制IRP2 mRNA的表达水平。结论atRA可通过影响IRP2 mRNA的表达,进而影响TFR mRNA、FnmRNA的水平来调节铁代谢。

Objective To study the influence of atRA on iron metabolism in cultured primary rat hepatocyte. Methods Rat primary hepatocytes were isolated by two-step in situ collagenase perfusion method by Seglen, and after that Cell viability was observed by 0.4 % trypan blue. And then primary hepatoeyte were treated into 6 wells plate with 0,0.5,1 and 50μmol/L atRA and DMEM contained 10 % fetal bovine serum. After 72h, IRP2 mRNA, TFR mRNA, Fnm RNA levels were measured by RT-PCR. Results VA deficiency can decrease the viability and function. Moreover hepatic IRP2 mRNA and TFRmRNA levels were increased by VA deficiency, which diminishing expression of Fn mRNA. Conclusion vitamin A deficiency can change cellular iron metabolism by inducing 1RP2-Fn-TFR pathway. AtRA supplementation inhibited the increase in IRP2 mRNA expression. Taken together, these results indicate that vitamin A deficiency can regulate iron metabolism by IRP2-TFR-Fn pathway.