高糖通过血清和糖皮质激素诱导蛋白激酶-1信号通路刺激人系膜细胞中纤连蛋白的合成

[High glucose stimulates synthesis of fibronectin in human mesangial cells via serum and glucocorticoid induced protein kinase-1 signaling pathway].

作者信息

Wang Quan-sheng, Zhang Xiao-li, Wang Yu-mei, Zhang A-li, Deng An-guo, Zhu Zhong-hua, Feng Yu-xi

机构信息

Renal Division, Union Hospital, Tongji Medical College, Huazhong Science and Technology University, Wuhan 430022, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2005 Jun 22;85(23):1591-5.

PMID:16185523

Abstract

OBJECTIVE

To investigate the role of serum and glucocorticoid induced kinase-1 (SGK(1)) pathways in fibronectin (FN) synthesis in human mesangial cell (HMC) under high glucose condition and the mechanism by which SGK(1) contributes to glomerulosclerosis in diabetic nephropathy (DN).

METHODS

HMCs were cultured and transfected with (P)IRES2-EGFP-(S422D) SGK(1) mutant (SD), plasmid containing SGK(1) dominant activation mutant, or blank plasmid. Non-transfected HMCs were used as control group. Then the HMCs were divided into 6 groups: transfected with SD + high glucose (SD-HG, 25 mmol/L D-glucose), transfected with FP + high glues (FP + HG), non-transfected + high glucose (NT-HG), transfeted with SD + normal glucose (SD-NG, 5.5 mmol/L D-glucose), transfected with FP + normal glues (FP + NG), and non-transfected + normal glucose (NT-NG). Eight hours after the glucose stimulation, RT-PCR was used to examine the SGK(1) mRNA expression and fibronectin (FN). Western blotting was used to detect the fibronectin (FN) protein expression.

RESULTS

The SGK(1) mRNA expression of the SD + HG group was 0.709, significantly higher than those of the FP + HG and NT + HP groups (0.497 and 0.491, both P < 0.01). The SGK(1) protein expression of the SD + HG group was 1,178,497, significantly higher than those of the FP + HG and NT + HP groups (193,875 and 195,597 respectively, both P < 0.01). The FN mRNA expression of the SD + HG group was 0.749, significantly higher than those of the FP + HG and NT + HP groups (0.463 and 0.475 respectively, both P < 0.01). The FN protein expression of the SD + HG group was 659,550, significantly higher than those of the FP + HG and NT + HG groups (342,354 and 340,428 respectively, both P < 0.01). There were not significant differences in the expressions of FN mRNA and protein among different NG groups.

CONCLUSION

SGK(1) may be involved in the signal transduction leading to the increase of fibronectin production in DN and therefore may play an active part in glomerulosclerosis in DN.

摘要

目的

探讨血清和糖皮质激素诱导激酶-1（SGK(1)）通路在高糖条件下人系膜细胞（HMC）中纤连蛋白（FN）合成中的作用，以及SGK(1)在糖尿病肾病（DN）肾小球硬化中的作用机制。

方法

培养HMC并分别用（P）IRES2-EGFP-（S422D）SGK(1)突变体（SD）、含SGK(1)显性激活突变体的质粒或空白质粒进行转染。未转染的HMC作为对照组。然后将HMC分为6组：转染SD + 高糖（SD-HG，25 mmol/L D-葡萄糖）、转染FP + 高糖（FP + HG）、未转染 + 高糖（NT-HG）、转染SD + 正常糖（SD-NG，5.5 mmol/L D-葡萄糖）、转染FP + 正常糖（FP + NG）、未转染 + 正常糖（NT-NG）。葡萄糖刺激8小时后，采用RT-PCR检测SGK(1) mRNA表达及纤连蛋白（FN）。采用蛋白质印迹法检测纤连蛋白（FN）蛋白表达。

结果

SD + HG组的SGK(1) mRNA表达为0.709，显著高于FP + HG组和NT + HP组（分别为0.497和0.491，均P < 0.01）。SD + HG组的SGK(1)蛋白表达为1,178,497，显著高于FP + HG组和NT + HP组（分别为193,875和195,597，均P < 0.01）。SD + HG组的FN mRNA表达为0.749，显著高于FP + HG组和NT + HP组（分别为0.463和0.475，均P < 0.01）。SD + HG组的FN蛋白表达为659,550，显著高于FP + HG组和NT + HG组（分别为342,354和340,428，均P < 0.01）。不同NG组之间FN mRNA和蛋白表达无显著差异。