文章摘要
左志华,游 娜,徐家蓉,蒋秀琴,缪 珩,鲁一兵.高糖状态下肾小管上皮细胞肌肌醇加氧酶表达变化对TGF-β1表达的影响[J].南京医科大学学报,2009,29(8):1089~1094
高糖状态下肾小管上皮细胞肌肌醇加氧酶表达变化对TGF-β1表达的影响
Effects of the abnormal expression of myo-inositol oxygenase (MIOX) on the expression of TGF-β1 in rat renal tubular epithelial cells under high glucose
投稿时间:2009-03-27  
DOI:10.7655
中文关键词: 肌肌醇加氧酶  肾小管上皮细胞  转化生长因子-β1  表达
英文关键词: myo-inositol oxygenase  renal tubular epithelial cells  TGF-β1  gene expression
基金项目:2007年江苏省高校自然科学基础研究面上项目(07KJD320141)
作者单位
左志华 南京医科大学第二附属医院内分泌科,江苏 南京 210011 
游 娜  
徐家蓉  
蒋秀琴  
缪 珩  
鲁一兵  
摘要点击次数: 2202
全文下载次数: 283
中文摘要:
      目的:探讨高糖状态下肌肌醇加氧酶(MIOX)表达变化对肾小管上皮细胞TGF-β1表达的影响?方法:①在体外培养大鼠肾小管上皮细胞NRK-52E,应用不同浓度高糖刺激NRK-52E细胞48 h,观察MIOX?TGF-β1 mRNA和蛋白表达的改变;②观测不同浓度TGF-β1刺激NRK-52E细胞48 h后MIOX mRNA和蛋白表达的变化;③应用针对MIOX基因的ASODN干涉NRK-52E细胞抑制MIOX基因表达后再用高糖刺激,观测其对TGF-β1 mRNA和蛋白表达的影响?结果:①不同浓度(15?30 mmol/L)高糖刺激NRK-52E细胞48 h后MIOX?TGF-β1 mRNA和蛋白表达增加;②不同浓度(1?5?10?25 mmol/L)TGF-β1刺激NRK-52E细胞48 h后MIOX mRNA和蛋白表达无明显变化;③针对MIOX基因的ASODN干涉NRK-52E细胞使MIOX基因表达下降,同时NRK-52E细胞TGF-β1 mRNA和蛋白的表达也下降?结论:本研究结果提示,高糖刺激肾小管上皮细胞能使MIOX?TGF-β1 mRNA和蛋白表达增加,而且MIOX表达异常可能通过影响细胞因子TGF-β1的表达在糖尿病肾病中起作用?
英文摘要:
      Objective:To investigate the effects of the abnormal expression of myo-inositol oxygenase (MIOX) on the expression of TGF-β1 in rat renal tubular epithelial cells under high glucose. Methods:① In vitro, rat renal tubular epithelial cell line NRK-52E was maintained in dulbecco’s modified eagle’s medium (DMEM) containing 5.6 mmol/L glucose and 10% fetal bovine serum (FBS). Then the cells were treated with D-glucose at different concentrations (ranging from 15 to 30 mmol/L) for 48 h, D-mannitol served as control. Gene expressions were analyzed at mRNA level by semi-quantitative RT-PCR and at protein level by Western blot. ② The expressions of MIOX mRNA and protein were observed in NRK-52E cells treated with TGF-β1 at different concentrations (ranging from 1 to 25 mmol/L) for 48 h. ③ Transient transfection of the normal NRK-52E cells with anti-sense oligodeoxynucleotide (ASODN) for 8 h was performed to knockdown MIOX gene using LipofectamineTM 2000. The cells were then treated with D-glucose at concentration of 30 mmol/L for 48 h. Gene expressions were analyzed at mRNA level by RT-PCR and at protein level by Western blot. Results:① In vitro, the expression of MIOX mRNA and protein in NRK-52E cells was induced by high D-glucose in a dose-dependent manner. Paralleled increases were observed with respect to TGF-β1 (P < 0.05). ② There was no significant difference of the MIOX mRNA and protein levels in NRK-52E cells between each group after the stimulation of TGF-β1 at different concentrations (ranging from 1 to 25 mmol/L,P > 0.05). ③ Transient transfection of the normal NRK-52E cells with ASODN significantly attenuated the increase of TGF-β1 expression in normal NRK-52E cells induced by high D-glucose (P < 0.05). Conclusion:High glucose can induce the expression of MIOX and TGF-β1 gene in NRK-52E cells in a dose-dependent manner. MIOX might play a role in the progression of diabetic nephropathy in rat renal tubular epithelial cells through the activation of cytokine TGF-β1.
查看全文   查看/发表评论  下载PDF阅读器