Regulation of CCN2/CTGF and related cytokines in cultured peritoneal cells under conditions simulating peritoneal dialysis
Regulation of CCN2/CTGF and related cytokines in cultured peritoneal cells under conditions simulating peritoneal dialysis
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DOI：
10.1093/ndt/gfn524
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发表时间：
2009-02-01
影响因子：
6.1
通讯作者：
Lai, Kar Neng
中科院分区：
文献类型：
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作者：
Leung, Joseph C. K.;Chan, Loretta Y. Y.;Lai, Kar Neng
背景持续性非卧床腹膜透析（CAPD）是终末期肾功能衰竭的主要治疗方式。腹膜表现出与透析持续时间相关的病理变化。这些变化是由于腹膜暴露于葡萄糖含量高的非生理性腹膜透析液（PDS），并含有潜在毒性物质，包括葡萄糖降解产物（GDP）和晚期糖基化终产物（AGE）。结缔组织生长因子（CTGF/CCN 2）是CAPD中进行性纤维化和腹膜功能障碍的决定因素之一。在这项研究中，我们研究了CCN 2的表达及其调节腹膜驻留细胞使用细胞培养模型。通过定量PCR（qPCR）检测与各种PDS及其组分一起培养的人腹膜间皮细胞（HPMC）、人腹膜成纤维细胞（HPF）或内皮细胞系EA.hy926（EC）中转化生长因子-β（TGF-β）、CCN 2和血管内皮生长因子（VEGF）的表达。使用条件培养基转移系统检查在HPMC与HPF或EC之间的串扰下CCN 2合成的调节，其中将HPMC暴露于从与PDS及其组分一起孵育的HPF或EC获得的条件培养基。采用凝胶电泳迁移率变动分析（EMSA）和qPCR检测TGF-β、CCN 2和VEGF诱导转录因子以及白细胞介素6（IL-6）、基质金属肽酶9（MMP-9）和I型胶原表达的差异。PDS及其组分对HPMC、HPF和EC中TGF-β、CCN 2和VEGF的表达具有差异性调节作用。在与HPF条件培养基和EC条件培养基共同培养后，HPMC表达CCN 2的量显著增加。中和抗TGF-β抗体减少但不完全消除与HPF或EC条件培养基一起培养的HPMC中的CCN 2合成。CCN 2、TGF-β和VEGF激活了HPMC中不同的转录因子，导致了不同的生物学反应，包括IL-6、MMP-9和I型胶原mRNA的表达。PDS中的AGE和GDPs差异调节腹膜驻留细胞合成CCN 2。通过HPMC的CCN 2合成可以通过从HPF或EC释放的TGF-β进一步扩增。不同转录因子的差异激活和HPMC对CCN 2、TGF-β和VEGF的不同反应表明，这些细胞因子/生长因子对HPMC具有重叠和不同的作用。
Background. Continuous ambulatory peritoneal dialysis (CAPD) is a major treatment modality for end-stage renal failure. The peritoneal membrane exhibits pathological changes that correlate with the duration of dialysis. These changes are due to the exposure of the peritoneum to non-physiologic peritoneal dialysis solution (PDS) with a high glucose content, and containing potentially toxic substances including glucose degradation products (GDP) and advanced glycation end products (AGE). Connective tissue growth factor (CTGF/CCN2) is one of the determinants of progressive fibrosis and peritoneal membrane dysfunction in CAPD. In this study, we examined the CCN2 expression and its regulation in peritoneal resident cells using a cell culture model.Methods. The expression of transforming growth factor-beta (TGF-beta), CCN2 and vascular endothelial growth factor (VEGF) in human peritoneal mesothelial cells (HPMC), human peritoneal fibroblasts (HPF) or endothelial cell line EA.hy926 (EC) cultured with various PDS and their components was examined by quantitative PCR (qPCR). The modulation of CCN2 synthesis under the crosstalk between HPMC and HPF or EC was examined using a conditioned medium transfer system in which HPMC was exposed to conditioned media obtained from HPF or EC incubated with PDS and their components. The differential effects of TGF-beta, CCN2 and VEGF in inducing the expression of transcriptional factors as well as interleukin-6 (IL-6), matrix metallopeptidase 9 (MMP-9) and collagen I were examined by electrophoretic mobility-shift assay (EMSA) and qPCR.Results. PDS and their components differentially modulated the expression of TGF-beta, CCN2 and VEGF in HPMC, HPF and EC. The expression of CCN2 by HPMC was significantly increased after cultured with a HPF-conditioned medium and an EC-conditioned medium. Neutralizing anti-TGF-beta antibodies reduced but not completely abolished the CCN2 synthesis in HPMC cultured with the HPF- or EC-conditioned medium. CCN2, TGF-beta and VEGF activated distinct transcriptional factors in HPMC, which resulted in divergent biological responses in terms of IL-6, MMP-9 and collagen I mRNA expression.Conclusion. AGE and GDPs in PDS differentially regulate the synthesis of CCN2 by peritoneal resident cells. The CCN2 synthesis by HPMC can be further amplified by TGF-beta released from HPF or EC. The differential activation of different transcriptional factors and diverse response of HPMC towards CCN2, TGF-beta and VEGF suggest that these cytokines/growth factors have an overlapping and distinct role on HPMC.