Effects of peritoneal effluents on mesothelial cells in culture: cell proliferation and extracellular matrix regulation.
Effects of peritoneal effluents on mesothelial cells in culture: cell proliferation and extracellular matrix regulation.
复制标题
腹膜流出物对培养物间皮细胞的影响：细胞增殖和细胞外基质调节。
DOI：
10.1093/ndt/11.9.1803
复制
发表时间：
1996
期刊：
影响因子：
--
通讯作者：
R. Gusmano
中科院分区：
文献类型：
--
作者：
F. Perfumo;P. Altieri;M. L. Degl’Innocenti;G. Ghiggeri;G. Caridi;A. Trivelli;R. Gusmano
腹膜透析患者的腹膜纤维化是腹膜内和壁间隙中间质胶原积聚的结果。假设靶细胞的胶原蛋白表达可能受特定的腹膜内因子调节，尽管这些因子的存在尚未得到证实。我们评估了从六名接受腹膜透析的儿童中获得的无细胞腹膜流出物对体外几种间皮细胞功能的影响。人腹膜间皮细胞（MC）是从6名尿毒症儿童的网膜组织中获得的，这些儿童正在接受腹腔导管插入手术。汇合时的细胞用于测定细胞毒性（LDH释放）、活力（台盼蓝）、增殖（3 H-胸苷掺入）、胶原蛋白表达（3 H-脯氨酸掺入，SDS-Page）和mRNA（斑点印迹）。进行了一系列初步实验，以确定透析过程中哪种连续液体收集引起最大变化;这揭示了长停滞期流出物的最大影响。暴露于从4例急性腹膜炎患者获得的腹膜流出物诱导细胞形态的显着变化，刺激（3 H）-胸苷掺入DNA的300%和上调III型胶原蛋白的表达和转录（6倍增量COL 3A 1 mRNA）。无腹膜炎患者的腹膜流出物诱导了细胞增殖（+100%）和胶原蛋白表达的定性但非定量可比变化。为了在体外重现腹膜流出物的作用，我们将间皮细胞暴露于感染的腹膜流出物中存在的各种细胞因子，即IL-2，TGF β和TNF α;在任何情况下，我们都没有发现细胞增殖的刺激。最后，TGF β而不是TNF α或IL 2上调这些细胞的胶原合成。这些发现表明无细胞腹膜流出物对间皮细胞功能的直接影响，包括刺激间质胶原蛋白表达。所有这些变化在暴露于急性腹膜炎期间收集的流出物时更加明显，这表明复发性腹膜感染与胶原沉积（腹膜纤维化的最典型前体）之间存在联系。
Peritoneal fibrosis in patients on peritoneal dialysis is the result of interstitial collagen accumulation within the peritoneal membrane and in mural spaces. Hypothetically, collagen expression by target cells may be regulated by specific endoperitoneal factors, though the existence of such factors has not yet been demonstrated. We evaluated the effects of cell-free peritoneal effluents obtained from six children undergoing peritoneal dialysis on several mesothelial cell functions in vitro. Human peritoneal mesothelial cells (MC) were obtained from the omental tissue of six uraemic children who were undergoing surgery for insertion of a peritoneal catheter. Cells at confluence were utilized to determine cytotoxicity (LDH release), viability (trypan blue), proliferation (3H-thymidine incorporation), collagen expression (3H-proline incorporation, SDS-Page) and mRNA (dot-blot). A preliminary series of experiments, was undertaken to define which of the successive fluid collections during a dialytic procedures induces the greatest changes; this revealed maximal effects of the effluent from the long stasis period. Exposure to peritoneal effluents obtained from four patients with acute peritonitis induced marked changes in cell morphology, stimulated by (3H)-thymidine incorporation into DNA by 300% and upregulated the expression and transcription of type III collagen (6-fold increment in COL3A1 mRNA). Qualitatively but not quantitatively comparable changes in cell proliferation (+100%) and collagen expression were induced by peritoneal effluents from patients without peritonitis. In an effort to reproduce the effect of peritoneal effluents in vitro, we exposed mesothelial cells to various cytokines putatively present in infected peritoneal effluents, namely IL-2, TGF beta and TNF alpha; in no case did we find stimulation of cell proliferation. Finally TGF beta but not TNF alpha or IL2 upregulated collagen synthesis by these cells. These findings demonstrate a direct influence of cell-free peritoneal effluents on mesothelial cell functions, including stimulation of interstitial collagen expression. All these changes were more evident upon exposure to effluents collected during acute peritonitis, which suggests a link between recurrent peritoneal infection and collagen deposition, the most typical precursor of peritoneal fibrosis.