摘要
目的观察红景天甙(salidroside,Sal)对氯化钴(cobaltous chloride,CoCl2.6 H2O,Co)诱导的低氧状态下正常大鼠近端肾小管上皮细胞(normal rat kidney tubular epithelia cell,NRK52E)转分化的影响并分析其可能的机制。方法体外培养NRK52E细胞,分为正常对照组,氯化钴低氧组,10、50、100μmol/L不同浓度红景天甙组。观察低氧标志蛋白低氧诱导因子-1α(hypoxia inducible foctor-1α,HIF-1α)的表达。倒置相差显微镜观察细胞形态变化;荧光免疫及细胞免疫法检测NRK52E细胞α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)表达情况;RT-PCR检测NRK52E细胞α-SMA、转化生长因子-β1(transforming growth factor-β1,TGF-β1)mRNA的表达;Western blot蛋白印迹检测HIF-1α和α-SMA蛋白的表达;酶联免疫吸附法检测细胞上清液胶原Ⅰ(collagenⅠ,Col-Ⅰ)和纤维粘连蛋白(fibronectin,FN)的含量。结果100μmol/L Co诱导NRK52E细胞表达HIF-1α。Co组细胞明显肥大、拉长,呈肌成纤维细胞外观。不同浓度Sal组细胞形态介于正常与Co组之间。Co组α-SMA基因、蛋白和TGF-β1mRNA表达量较对照组升高(P<0.05),不同浓度Sal组α-SMA基因、蛋白和TGF-β1mRNA表达量较Co组减少(P<0.05)。Co组细胞上清液Col-Ⅰ和FN的含量较对照组增加(P<0.05),不同浓度Sal组Col-Ⅰ和FN的含量明显下降(P<0.05),以高剂量组最为明显。结论Sal可在一定程度上改善Co诱导的低氧状态和细胞转分化,减少细胞外基质的增加。其机制可能是通过抑制TGF-β1和HIF-1α的表达实现的。
Objective To investigate the effect and possible mechanism of salidroside on the transdifferentiation of normal rat kidney tubular epithelia cells （NRK52E） under cobaltous chloride （Co） induced hypoxic status. Methods Cultured NRK52E cells were divided into control group, Co group and Co plus salidroside treatment groups at a dosage of 10 μmol/L, 50 μmol/L, and 100 μmol/L. Hypoxia-inducible factor-1α （HIF-1α）, a master regulator of oxygen homeostasis was measured as a marker of hypoxic status. Morphologic alteration of cells was observed by inverted phase contrast microscope. The expression of α-SMA in NRK52E cells was detected by fluorescent immunocytochemistry （FICC） and immunohistochemistry （IHC）. The a-SMA and TGF-β1 mRNA were assessed using reverse transcription-polymerase chain reaction （RT-PCR）. The expressions of HIF-1α and α-SMA protein were detected by Western blot analyses. The enzyme-linked immunosorbent assay was performed to detect collagen I （Col- l ） and fibronectin （FN） in the supernatant. Results The expression of HIF- la in NRK52E cells was induced by 100μmol/L of Co in vitro. Co induced transdifferentiation of NRK52E cells, showing fibroblast-like in morphology. Salidroside partly blocked morphologic transformation of tubular epithelial cells. Salidroside decreased the expressions of α-SMA protein and mRNA and TGF-β1 mRNA significantly（P〈 0.05）, although they were still higher than the controls（P〈0.05）. Salidroside, especially in high dosage, inhibited the increase in Col- l and FN induced by Co （P〈0. 05）. Conclusion Hypoxia can induce tubular epithelial- myofibroblast transdifferentiation （TEMT）. Salidroside improves Co-induced hypoxic status and inhibits TEMT possibly through reducing Col I and FN in NRK52E cells.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2010年第1期43-48,共6页
Journal of Sichuan University(Medical Sciences)
基金
教育部博士点基金(项目编号20060610063)
成都市卫生局科研重点项目(项目编号07-17)资助
关键词
低氧
红景天甙
肾小管上皮细胞转分化
低氧诱导因子-1α转化生长因子-β1
Hypoxia Salidroside Tubular epithelial-myofibroblast transdifferentiation（TEMT） Hypoxia induced foctor-1 （HIF-1） Transforming growth factor-β1 （TGF-β1）
