转化生长因子-β信号在持续性非卧床腹膜透析腹膜硬化中的作用
Involvement of TGF-beta signal for peritoneal sclerosing in continuous ambulatory peritoneal dialysis.
作者信息
Naiki Yoshito, Maeda Yasuhiro, Matsuo Koki, Yonekawa Satoru, Sakaguchi Mika, Iwamoto Ichiro, Hasegawa Hirofumi, Kanamaru Akihisa
机构信息
Department of Internal Medicine, Division of Hematology, Nephrology and Rheumatology, Kinki University School of Medicine, Osaka, Japan.
出版信息
J Nephrol. 2003 Jan-Feb;16(1):95-102.
BACKGROUND
Functional failure of the peritoneal membrane is the most serious problem in long-term continuous ambulatory peritoneal dialysis (CAPD). Transforming growth factor-beta (TGF-ss) is one of the key mediators of fibrosis in some organs, and is thought to be involved in peritoneal alterations. In this study, we examined the role of TGF-beta1/TGF-ss receptors for human peritoneal mesothelial cells (HPMCs) and fibroblasts, and their interactions in CAPD patients.
METHODS
HPMCs were cultured for 48 h in a medium containing normal- dose glucose (7 mM), high-dose glucose (30 mM) and mannitol as an osmotic agent, equal to 30 mM glucose. Cell proliferation was observed using the Tetra Color One assay. The concentration of TGF-beta1 in culture supernatants was measured by enzyme-linked immunosorbent assay (ELISA). The expression of TGF-ss receptor types I and II was observed by flow cytometry. HPMCs and fibroblasts were co-cultured and assayed using transwell inserts in order to identify the effects of the high-concentration glucose solution.
RESULTS
HPMC proliferation was inhibited by the high concentration of glucose but not by mannitol. The inhibition was abrogated by the neutralizing antibody for TGF-beta1. TGF-beta1 was induced by a high concentration of glucose but not by mannitol. The expression of both TGF-ss receptors was augmented in culture with the high concentration of glucose but not with mannitol. In the co-culture assay, the number of HPMCs was decreased and fibroblasts were significantly increased in culture with the high concentration of glucose.
CONCLUSIONS
A high concentration of glucose induced a large amount of TGF-beta1 and enhanced the expression of TGF-ss receptors. HPMCs were sensitive to TGF-beta1 in response to a high concentration of glucose. These data suggest that TGF-beta1 from HPMCs exposed to a high concentration of glucose down-regulates the proliferation of HPMCs and accelerates peritoneal fibrosis.
背景
腹膜功能衰竭是长期持续性非卧床腹膜透析（CAPD）中最严重的问题。转化生长因子-β（TGF-β）是某些器官纤维化的关键介质之一，被认为与腹膜改变有关。在本研究中，我们研究了TGF-β1/TGF-β受体对人腹膜间皮细胞（HPMC）和成纤维细胞的作用，以及它们在CAPD患者中的相互作用。
方法
将HPMC在含有正常剂量葡萄糖（7 mM）、高剂量葡萄糖（30 mM）和作为渗透剂的甘露醇（相当于30 mM葡萄糖）的培养基中培养48小时。使用四色一步法观察细胞增殖。通过酶联免疫吸附测定（ELISA）测量培养上清液中TGF-β1的浓度。通过流式细胞术观察TGF-β受体I型和II型的表达。将HPMC和成纤维细胞共培养，并使用Transwell小室进行测定，以确定高浓度葡萄糖溶液的作用。
结果
高浓度葡萄糖抑制HPMC增殖，但甘露醇无此作用。TGF-β1中和抗体可消除这种抑制作用。高浓度葡萄糖诱导TGF-β1产生，但甘露醇无此作用。高浓度葡萄糖培养时，TGF-β受体的表达均增加，但甘露醇培养时无此现象。在共培养试验中，高浓度葡萄糖培养时HPMC数量减少，成纤维细胞显著增加。
结论
高浓度葡萄糖诱导大量TGF-β1产生并增强TGF-β受体的表达。HPMC对高浓度葡萄糖诱导的TGF-β1敏感。这些数据表明，暴露于高浓度葡萄糖的HPMC产生的TGF-β1下调HPMC的增殖并加速腹膜纤维化。
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