活性氧介导高糖负荷致人系膜细胞癌胚纤维连接蛋白的表达

摘要

BACKGROUND: Researches showed that oncofetal fibronectin is an important indicator of new synthetic extracellular matrix. But studies of relationship between oxidative stress in high glucose environment and oncofetal fibronection are rarely reported.OBJECTIVE: To investigate the effect of high glucose on the expression of reactive oxygen species (ROS) and oncofetal fibronectin mRNA in human mesangial cells.METHODS: The cultured mesangial cells were divided into following groups: normal control group (5 mmol/L D-glucose), osmotic control group (5 mmol/L D-glucose+20 mmol/L L-glucose) and high glucose group (25 mmol/L D-glucose); α-lipoic acid intervention group containing high glucose+α-lipoic acid 50 group (25 mmol/L D-glucose+50 μmol/L α-lipoic acid), high glucose+α-lipoic acid 100 group (25 mmol/L D-glucose+100 μmol/L α-lipoic acid) and high glucose+α-lipoic acid 200 group (25 mmol/L D-glucose+200 μmol/L α-lipoic acid). The expression of oncofetal fibronectin mRNA was detected by reverse transcription-PCR. ROS levels were tested by fluorescence microscopy and fluorescence microplate reader. RESULTS AND CONCLUSION: High glucose could promote the production of ROS and increase the expression of oncofetal fibronectin mRNA in the mesangial cells. Α-lipoic acid could significantly reduce the level of ROS and expression of oncofetal fibronectin mRNA in the mesangial cells under high glucose loaded, which showed a concentration-dependent manner. It is indicated that ROS mediated high glucose can induce the expression of oncofetal fibronectin in the mesangial cells, but this effect can be partially reversed by α-lipoic acid.%背景:研究发现,癌胚纤维连接蛋白系新合成细胞外基质的重要标志,而高糖环境氧化应激与癌胚纤维连接蛋白的关系尚未见报道.目的:观察高糖作用对人系膜细胞活性氧和癌胚纤维连接蛋白mRNA表达的影响.方法:将培养的系膜细胞分为以下各组:正常对照组(5 mmol/L D-葡萄糖);渗透压对照组(5 mmol/L D-葡萄糖+ 20 mmol/L L-葡萄糖);高糖组(25 mmol/L D-葡萄糖);α-硫辛酸干预组,分为高糖+LA50组(25 mmol/L D-葡萄糖+ 50 μmol/L α-硫辛酸)、高糖+LA100组(25 mmol/L D-葡萄糖+100 μmol/L α-硫辛酸)、高糖+LA200组(25 mmol/L D-葡萄糖+ 200 μmol/L α-硫辛酸).以RT-PCR法检测癌胚纤维连接蛋白mRNA表达水平,荧光显微镜和荧光酶标仪测定细胞内活性氧水平.结果与结论:高糖促进系膜细胞活性氧的产生,亦增加癌胚纤维连接蛋白mRNA的表达,α-硫辛酸可显著降低高糖负荷下细胞内活性氧水平,同时减少癌胚纤维连接蛋白mRNA的表达,且呈浓度依赖性,提示活性氧介导高糖负荷致人系膜细胞癌胚纤维连接蛋白的表达,α-硫辛酸可部分逆转这一效应.