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甘露糖结合凝集素对高糖培养的人肾小球内皮细胞中转化生长因子-β1和核因子-κB表达的影响

[Influence of mannose binding lectin to expression of TGF-beta1 and NF-kappaB in high concentration of glucose cultured human renal glomerular endothelial cells].

作者信息

Qiu Hong-Yu, Fan Wen-Xing, Zuo Chuan, Huang Song-Min, Liu Fang, Tang Wan-Xin

机构信息

Department of Nephrology, West China Hospital, Sichuan University, Chengdu 610041, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2011 Sep;42(5):604-9.

Abstract

OBJECTIVE

To investigate the effect of mannose binding lectin (MBL) complement pathway on expression of transforming growth factor-beta1 (TGF-beta1) and NF-kappaB in cultured human renal glomerular endothelial cells (HRGECs) stimulated by high concentration of glucose.

METHODS

Human glomerular endothelial cells in culture were randomly divided into 5 groups according to different managements: normal concentration of glucose as controlled group, MBL + normal concentration of glucose group, high concentration of glucose, MBL + high glucose and MBL + high glucose + MBL blocker respectively. Flow cytometry was used to detect the depositions of MBL and C3 on the surfaces of HRGECs. Real-time PCR method was used to detect the mRNA levels of TGF-beta1. Human TGF-beta1 ELISA kit was used to detect the concentration of TGF-beta1 in supernatant fluid. ESMA was used to detect the activity of NF-kappaB in HRGECs.

RESULTS

Compared with the normal glucose group and high glucose group, the depositions of MBL, C3 were apparently increased in MBL + high glucose group (P < 0.05). Expression of TGF-beta1 were significantly higher (P < 0.05) in MBL + high concentration of glucose groups than the normal glucose group and the high concentration of glucose group. Compared with the high glucose group, the activity of NF-kappaB in HRGECs was apparently increased in MBL + high glucose group, which could be significantly downregulated by MBL blocking antibody.

CONCLUSION

High concentration of glucose can increase the expression of TGF-beta1 of cultured human glomerular endothelial cells. At the same time, high glucose together with MBL can up regulate the expression of TGF-beta1 and the activity of NF-kappaB in HRGECs.

摘要

目的

探讨甘露糖结合凝集素(MBL)补体途径对高糖刺激的人肾小球内皮细胞(HRGECs)中转化生长因子-β1(TGF-β1)和核因子-κB(NF-κB)表达的影响。

方法

将培养的人肾小球内皮细胞根据不同处理随机分为5组:正常葡萄糖浓度组作为对照组、MBL+正常葡萄糖浓度组、高糖组、MBL+高糖组和MBL+高糖+MBL阻断剂组。采用流式细胞术检测HRGECs表面MBL和C3的沉积情况。采用实时荧光定量PCR法检测TGF-β1的mRNA水平。使用人TGF-β1 ELISA试剂盒检测上清液中TGF-β1的浓度。采用电泳迁移率变动分析(EMSA)检测HRGECs中NF-κB的活性。

结果

与正常葡萄糖组和高糖组相比,MBL+高糖组MBL、C3的沉积明显增加(P<0.05)。MBL+高浓度葡萄糖组TGF-β1的表达显著高于正常葡萄糖组和高浓度葡萄糖组(P<0.05)。与高糖组相比,MBL+高糖组HRGECs中NF-κB的活性明显增加,而MBL阻断抗体可使其显著下调。

结论

高浓度葡萄糖可增加培养的人肾小球内皮细胞中TGF-β1的表达。同时,高糖联合MBL可上调HRGECs中TGF-β1的表达及NF-κB的活性。

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