白细胞介素-1β诱导人近端肾小管细胞损伤、α-平滑肌肌动蛋白表达和纤连蛋白生成。

Interleukin-1beta induces human proximal tubule cell injury, alpha-smooth muscle actin expression and fibronectin production.

作者信息

Vesey David A, Cheung Catherine W Y, Cuttle Leila, Endre Zoltan A, Gobé Glenda, Johnson David W

机构信息

Department of Renal Medicine, Princess Alexandra Hospital, Woolloongabba, Brisbane, Queensland 4102, Australia.

出版信息

Kidney Int. 2002 Jul;62(1):31-40. doi: 10.1046/j.1523-1755.2002.00401.x.

DOI:10.1046/j.1523-1755.2002.00401.x

PMID:12081561

Abstract

BACKGROUND

Tubulointerstitial lesions, characterized by tubular injury, interstitial fibrosis and the appearance of myofibroblasts, are the strongest predictors of the degree and progression of chronic renal failure. These lesions are typically preceded by macrophage infiltration of the tubulointerstitium, raising the possibility that these inflammatory cells promote progressive renal disease through fibrogenic actions on resident tubulointerstitial cells. The aim of the present study, therefore, was to investigate the potentially fibrogenic mechanisms of interleukin-1beta (IL-1beta), a macrophage-derived pro-inflammatory cytokine, on human proximal tubule cells (PTC).

METHODS

Confluent, quiescent, passage 2 PTC were established in primary culture from histologically normal segments of human renal cortex (N = 11) and then incubated in serum- and hormone-free media supplemented with either IL-1beta (0 to 4 ng/mL) or vehicle (control).

RESULTS

IL-1beta significantly enhanced fibronectin secretion by up to fourfold in a time- and concentration-dependent fashion. This was accompanied by significant (2.5- to 6-fold) increases in alpha-smooth muscle actin (alpha-SMA) expression, transforming growth factor beta (TGF-beta1) secretion, nitric oxide (NO) production, NO synthase 2 (NOS2) mRNA and lactate dehydrogenase (LDH) release. Cell proliferation was dose-dependently suppressed by IL-1beta. NG-methyl-l-arginine (L-NMMA; 1 mmol/L), a specific inhibitor of NOS, blocked NO production but did not alter basal or IL-1beta-stimulated fibronectin secretion. In contrast, a pan-specific TGF-beta neutralizing antibody significantly blocked the effects of IL-1beta on PTC fibronectin secretion (IL-1beta, 268.1 +/- 30.6 vs. IL-1beta+alphaTGF-beta 157.9 +/- 14.4%, of control values, P < 0.001) and DNA synthesis (IL-1beta 81.0 +/- 6.7% vs. IL-1beta+alphaTGF-beta 93.4 +/- 2.1%, of control values, P < 0.01).

CONCLUSION

IL-1beta acts on human PTC to suppress cell proliferation, enhance fibronectin production and promote alpha-smooth muscle actin expression. These actions appear to be mediated by a TGF-beta1 dependent mechanism and are independent of nitric oxide release.

摘要

背景

以肾小管损伤、间质纤维化和成肌纤维细胞出现为特征的肾小管间质病变是慢性肾衰竭程度和进展的最强预测指标。这些病变通常先于肾小管间质巨噬细胞浸润，这增加了这些炎症细胞通过对肾小管间质驻留细胞的促纤维化作用促进进行性肾病的可能性。因此，本研究的目的是探讨巨噬细胞衍生的促炎细胞因子白细胞介素-1β（IL-1β）对人近端肾小管细胞（PTC）的潜在促纤维化机制。

方法

从人肾皮质组织学正常节段原代培养建立汇合、静止、第2代PTC（n = 11），然后在补充有IL-1β（0至4 ng/mL）或载体（对照）的无血清和无激素培养基中孵育。

结果

IL-1β以时间和浓度依赖性方式显著增强纤连蛋白分泌达四倍。这伴随着α-平滑肌肌动蛋白（α-SMA）表达、转化生长因子β（TGF-β1）分泌、一氧化氮（NO）产生、NO合酶2（NOS2）mRNA和乳酸脱氢酶（LDH）释放显著增加（2.5至6倍）。IL-1β剂量依赖性抑制细胞增殖。NOS特异性抑制剂NG-甲基-L-精氨酸（L-NMMA；1 mmol/L）阻断NO产生，但不改变基础或IL-1β刺激的纤连蛋白分泌。相反，泛特异性TGF-β中和抗体显著阻断IL-1β对PTC纤连蛋白分泌（IL-1β，对照值的268.1±30.6 vs. IL-1β+αTGF-β 157.9±14.4%，P < 0.001）和DNA合成（IL-β 81.0±6.7% vs. IL-1β+αTGF-β 93.4±2.1%，对照值，P < 0.01）的作用。