Vesey David A, Cheung Catherine, Cuttle Leila, Endre Zoltan, Gobe Glenda, Johnson David W
Department of Renal Medicine, Princess Alexandra Hospital, Brisbane, Qld, Australia.
J Lab Clin Med. 2002 Nov;140(5):342-50. doi: 10.1067/mlc.2002.128468.
One of the hallmarks of progressive renal disease is the development of tubulointerstitial fibrosis. This is frequently preceded by macrophage infiltration, raising the possibility that macrophages relay fibrogenic signals to resident tubulointerstitial cells. The aim of this study was to investigate the potentially fibrogenic role of interleukin-1beta (IL-1beta), a macrophage-derived inflammatory cytokine, on cortical fibroblasts (CFs). Primary cultures of human renal CFs were established and incubated for 24 hours in the presence or absence of IL-1beta. We found that IL-1beta significantly stimulated DNA synthesis (356.7% +/- 39% of control, P <.003), fibronectin secretion (261.8 +/- 11% of control, P <.005), collagen type 1 production, (release of procollagen type 1 C-terminal-peptide, 152.4% +/- 26% of control, P <.005), transforming growth factor-beta (TGF-beta) secretion (211% +/- 37% of control, P <.01), and nitric oxide (NO) production (342.8% +/- 69% of control, P <.002). TGF-beta (1 ng/mL) and the phorbol ester phorbol 12-myristate 13-acetate (PMA, 25 nmol/L) produced fibrogenic effects similar to those of IL-1beta. Neither a NO synthase inhibitor (N(G)-methyl-l-arginine, 1 mmol/L) nor a protein kinase C (PKC) inhibitor (bis-indolylmaleimide 1, 1 micromol/L) altered the enhanced level of fibronectin secretion or DNA synthesis seen in response to IL-1beta treatment. However, addition of a TGF-beta-neutralizing antibody significantly reduced IL-1beta-induced fibronectin secretion (IL-1beta + IgG, 262% +/- 72% vs IL-1beta + alphaTGF-beta 156% +/- 14%, P <.02), collagen type 1 production (IL-1beta + IgG, 176% +/- 28% vs IL-1beta + alphaTGF-beta, 120% +/- 14%, P <.005) and abrogated IL-1beta-induced DNA synthesis (245% +/- 49% vs 105% +/- 21%, P <.005). IL-1beta significantly stimulated CF DNA synthesis and production of fibronectin, collagen type 1, TGFbeta, and NO. The fibrogenic and proliferative action of IL-1beta on CF appears not to involve activation of PKC or production of NO but is at least partly TGFbeta-dependent.
进行性肾病的一个标志是肾小管间质纤维化的发展。这通常先于巨噬细胞浸润，这增加了巨噬细胞将促纤维化信号传递给肾小管间质驻留细胞的可能性。本研究的目的是研究巨噬细胞衍生的炎性细胞因子白细胞介素-1β（IL-1β）对皮质成纤维细胞（CFs）的潜在促纤维化作用。建立人肾CFs原代培养物，并在有或无IL-1β的情况下孵育24小时。我们发现IL-1β显著刺激DNA合成（对照的356.7%±39%，P<.003）、纤连蛋白分泌（对照的261.8±11%，P<.005）、I型胶原产生（I型前胶原C末端肽释放，对照的152.4%±26%，P<.005）、转化生长因子-β（TGF-β）分泌（对照的211%±37%，P<.01）和一氧化氮（NO）产生（对照的342.8%±69%，P<.002）。TGF-β（1 ng/mL）和佛波酯佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA，25 nmol/L）产生与IL-1β相似的促纤维化作用。一氧化氮合酶抑制剂（N(G)-甲基-L-精氨酸，1 mmol/L）和蛋白激酶C（PKC）抑制剂（双吲哚基马来酰亚胺1，1 μmol/L）均未改变IL-1β处理后纤连蛋白分泌或DNA合成增强的水平。然而，添加TGF-β中和抗体显著降低了IL-1β诱导的纤连蛋白分泌（IL-1β+IgG，262%±72%对IL-1β+αTGF-β 156%±14%，P<.02）、I型胶原产生（IL-1β+IgG，176%±28%对IL-1β+αTGF-β，120%±14%，P<.005），并消除了IL-1β诱导的DNA合成（245%±49%对105%±21%，P<.005）。IL-1β显著刺激CF DNA合成以及纤连蛋白、I型胶原、TGFβ和NO的产生。IL-1β对CF的促纤维化和增殖作用似乎不涉及PKC的激活或NO的产生，但至少部分依赖于TGFβ。
