Costalonga Elerson C, de Freitas Luiza J, Aragone Deise da S P, Silva Filipe M O, Noronha Irene L
Laboratory of Cellular, Genetic, and Molecular Nephrology, Renal Division, University of São Paulo Medical School, São Paulo, Brazil.
Center for Cellular and Molecular Studies and Therapy (NETCEM), University of São Paulo Medical School, São Paulo, Brazil.
PLoS One. 2017 Sep 5;12(9):e0184302. doi: 10.1371/journal.pone.0184302. eCollection 2017.
Progressive fibrous thickening of the peritoneal membrane is a complication of long-term peritoneal dialysis (PD). TGF-β/Smad pathway activation, inflammation, and neoangiogenesis play important roles in peritoneal membrane (PM) changes induced by PD. Recently, histone deacetilase inhibitors (HDACi) have shown anti-fibrotic and anti-inflammatory effects in different experimental models. These drugs prevent deacetylation of histones causing a loosen chromatin, which in turn induce the expression of some anti-fibrotic genes. In addition, acetylation may increase the activity of proteins involved in tissue fibrosis, such as Smad7. Here, we explored the effect of valproic acid (VPA), an HDACi, on the development of peritoneal fibrosis (PF) in rats.
PF was induced by daily intraperitoneal injections of 0.1% chlorhexidine gluconate (CG) for 15 consecutive days. Male Wistar rats (250-300 g) were divided into 3 groups: CONTROL, control rats receiving only vehicle; PF, peritoneal fibrosis induced in rats; PF+VPA, rats with PF treated with VPA (300 mg/kg/day by gavage). PF was assessed by Masson's trichrome staining. Inflammation and fibrosis-associated factors were assessed by immunohistochemistry, immunofluorescence, multiplex analysis, and qPCR.
Treatment with VPA significantly reduced PM thickness and the expression of myofibroblasts, besides preventing loss of ultrafiltration capacity of the PM. The upregulation of profibrotic factors (TGF-β, fibronectin, and Smad3) in the PF group was significantly ameliorated by VPA. VPA modulated the TGF/Smad pathway, inhibiting phosphorylated Smad3 expression and inducing an increased Smad7 expression in the FP+VPA group. The neoangiogenesis and the expression of proinflammatory cytokines (TNF-α, IL-1β, MCP-1) observed in the PF group was significantly reduced by VPA.
Our results indicate that VPA suppressed experimental PF through modulation of the TGF-β/Smad pathway. Interestingly, VPA treatment induced a higher expression of antifibrotic factors, such as Smad7. These results suggest that VPA may represent a potential strategy for treating long term PD complications.
腹膜进行性纤维增厚是长期腹膜透析(PD)的一种并发症。转化生长因子-β/信号转导和转录激活因子(TGF-β/Smad)通路激活、炎症和新生血管生成在PD诱导的腹膜变化中起重要作用。近来,组蛋白去乙酰化酶抑制剂(HDACi)在不同实验模型中已显示出抗纤维化和抗炎作用。这些药物可防止组蛋白去乙酰化,导致染色质疏松,进而诱导一些抗纤维化基因的表达。此外,乙酰化作用可能会增强参与组织纤维化的蛋白质(如Smad7)的活性。在此,我们探究了HDACi丙戊酸(VPA)对大鼠腹膜纤维化(PF)发展的影响。
通过每日腹腔注射0.1%葡萄糖酸氯己定(CG)连续15天诱导PF。将雄性大鼠(250 - 300克)分为3组:对照组,仅接受赋形剂的对照大鼠;PF组,诱导大鼠发生腹膜纤维化;PF + VPA组,用VPA治疗的PF大鼠(通过灌胃给予300毫克/千克/天)。通过Masson三色染色评估PF。通过免疫组织化学、免疫荧光、多重分析和定量聚合酶链反应(qPCR)评估炎症和纤维化相关因子。
VPA治疗除了防止腹膜超滤能力丧失外,还显著降低了腹膜厚度和成肌纤维细胞的表达。PF组中促纤维化因子(TGF-β、纤连蛋白和Smad3)的上调被VPA显著改善。VPA调节TGF/Smad通路,在PF + VPA组中抑制磷酸化Smad3表达并诱导Smad7表达增加。VPA显著降低了PF组中观察到的新生血管生成和促炎细胞因子(肿瘤坏死因子-α、白细胞介素-1β、单核细胞趋化蛋白-1)的表达。
我们的结果表明,VPA通过调节TGF-β/Smad通路抑制实验性PF。有趣的是,VPA治疗诱导了抗纤维化因子(如Smad7)的更高表达。这些结果表明,VPA可能是治疗长期PD并发症的一种潜在策略。