Background: Peritoneal fibrosis is the primary reason that patients with end-stage renal disease (ESRD) have to cease peritoneal dialysis. Peritonitis caused by Gram-negative bacteria such as Escherichia coli (E. coli...Background: Peritoneal fibrosis is the primary reason that patients with end-stage renal disease (ESRD) have to cease peritoneal dialysis. Peritonitis caused by Gram-negative bacteria such as Escherichia coli (E. coli) were on the rise. We had previously shown that matrine inhibited the formation of biofilm by E. coli. However, the role of matrine on the epithelial-mesenchymal transition (EMT) in peritoneal mesothelial cells under chronic inflammatory conditions is still unknown. Methods: We cultured human peritoneal mesothelial cells (HPMCs) with lipopolysaccharide (LPS) to induce an environment that mimicked peritonitis and investigated whether matrine could inhibit LPS-induced EMT in these cells. In addition, we investigated the change in expression levels of the miR-29b and miR-129-5p. Results: We found that 10 jjig/ml of LPS induced EMT in HPMCs. Matrine inhibited LPS-induced EMT in HPMCs in a dose-dependent manner. We observed that treatment with matrine increased the expression of E-cadherin (F= 50.993, P< 0.01), and decreased the expression of alpha-smooth muscle actin (F= 32.913, P < 0.01). Furthermore, we found that LPS reduced the expression levels of miR- 29b and miR-129-5P in HPMCs, while matrine promoted the expression levels of miR-29b and miR-129-5P. Conclusions: Matrine could inhibit LPS-induced EMT in HPMCs and reverse LPS inhibited expressions of miR-29 b and miR-129- 5P in HPMCs, ultimately reduce peritoneal fibrosis. These findings provide a potential theoretical basis for using matrine in the prevention and treatment of peritoneal fibrosis.展开更多
The peritoneum response to peritoneal dialysis can lead to fibrosis. The transforming growth factor-β1 （TGF-β1） plays an important role in regulating tissue repair and remodeling after injury. Excessive synthesis ...The peritoneum response to peritoneal dialysis can lead to fibrosis. The transforming growth factor-β1 （TGF-β1） plays an important role in regulating tissue repair and remodeling after injury. Excessive synthesis and deposition of matrix proteins by peritoneal mesothelial cells can lead to structural and functional changes in the peritoneal membrane, jeopardizing the long-term efficacy of peritoneal dialysis （PD）. Prolonged exposure to high glucose concentrations in PD fluid has been implicated as a major stimulus to matrix accumulation, through the induction of transforming growth factor-β1 （TGF-β1）.展开更多
目的:观察钙浓度变化对人腹膜间皮细胞上皮细胞-间充质细胞转分化的影响,探讨腹膜透析中腹膜纤维化的可能机制。方法:以人腹膜间皮细胞永生化细胞株(HMrSV5)为研究对象,予不同浓度钙(0.25、0.75、1.25、1.75、2.25mmol/L)处理细胞48h,...目的:观察钙浓度变化对人腹膜间皮细胞上皮细胞-间充质细胞转分化的影响,探讨腹膜透析中腹膜纤维化的可能机制。方法:以人腹膜间皮细胞永生化细胞株(HMrSV5)为研究对象,予不同浓度钙(0.25、0.75、1.25、1.75、2.25mmol/L)处理细胞48h,以不含钙处理组为对照。采用蛋白免疫印迹和间接免疫荧光染色方法检测E-钙黏蛋白(E-cadherin)、成纤维细胞特异蛋白(fibroblast specific protein,FSP1)和波形蛋白(Vimentin)的表达。结果:不含钙组HMrSV5细胞基本不表达E-cadherin,极低水平表达Vimentin和FSP1;而予含不同浓度钙离子处理48h后,HMrSV5中E-cadherin先呈剂量依赖性上调,1.25mmol/L组至顶峰,再呈剂量依赖性下降;而Vimentin和FSP1均在1.75mmol/L组HMrSV5中明显升高(P<0.05)并呈剂量依赖性上调。间接免疫荧光染色结果也显示,高钙组Vimentin和FSP1荧光强度比低钙组显著增强。结论:钙离子浓度的变化能诱导人腹膜间皮细胞E-cadherin、FSP1、Vimentin蛋白表达发生变化,提示钙离子可能是参与调控间皮细胞间充质转分化的一个重要因素。展开更多
基金grants from the National Natural Science Foundation of China (No. 81360111 and No. 81660133).
文摘Background: Peritoneal fibrosis is the primary reason that patients with end-stage renal disease (ESRD) have to cease peritoneal dialysis. Peritonitis caused by Gram-negative bacteria such as Escherichia coli (E. coli) were on the rise. We had previously shown that matrine inhibited the formation of biofilm by E. coli. However, the role of matrine on the epithelial-mesenchymal transition (EMT) in peritoneal mesothelial cells under chronic inflammatory conditions is still unknown. Methods: We cultured human peritoneal mesothelial cells (HPMCs) with lipopolysaccharide (LPS) to induce an environment that mimicked peritonitis and investigated whether matrine could inhibit LPS-induced EMT in these cells. In addition, we investigated the change in expression levels of the miR-29b and miR-129-5p. Results: We found that 10 jjig/ml of LPS induced EMT in HPMCs. Matrine inhibited LPS-induced EMT in HPMCs in a dose-dependent manner. We observed that treatment with matrine increased the expression of E-cadherin (F= 50.993, P< 0.01), and decreased the expression of alpha-smooth muscle actin (F= 32.913, P < 0.01). Furthermore, we found that LPS reduced the expression levels of miR- 29b and miR-129-5P in HPMCs, while matrine promoted the expression levels of miR-29b and miR-129-5P. Conclusions: Matrine could inhibit LPS-induced EMT in HPMCs and reverse LPS inhibited expressions of miR-29 b and miR-129- 5P in HPMCs, ultimately reduce peritoneal fibrosis. These findings provide a potential theoretical basis for using matrine in the prevention and treatment of peritoneal fibrosis.
文摘The peritoneum response to peritoneal dialysis can lead to fibrosis. The transforming growth factor-β1 （TGF-β1） plays an important role in regulating tissue repair and remodeling after injury. Excessive synthesis and deposition of matrix proteins by peritoneal mesothelial cells can lead to structural and functional changes in the peritoneal membrane, jeopardizing the long-term efficacy of peritoneal dialysis （PD）. Prolonged exposure to high glucose concentrations in PD fluid has been implicated as a major stimulus to matrix accumulation, through the induction of transforming growth factor-β1 （TGF-β1）.
文摘目的:观察钙浓度变化对人腹膜间皮细胞上皮细胞-间充质细胞转分化的影响,探讨腹膜透析中腹膜纤维化的可能机制。方法:以人腹膜间皮细胞永生化细胞株(HMrSV5)为研究对象,予不同浓度钙(0.25、0.75、1.25、1.75、2.25mmol/L)处理细胞48h,以不含钙处理组为对照。采用蛋白免疫印迹和间接免疫荧光染色方法检测E-钙黏蛋白(E-cadherin)、成纤维细胞特异蛋白(fibroblast specific protein,FSP1)和波形蛋白(Vimentin)的表达。结果:不含钙组HMrSV5细胞基本不表达E-cadherin,极低水平表达Vimentin和FSP1;而予含不同浓度钙离子处理48h后,HMrSV5中E-cadherin先呈剂量依赖性上调,1.25mmol/L组至顶峰,再呈剂量依赖性下降;而Vimentin和FSP1均在1.75mmol/L组HMrSV5中明显升高(P<0.05)并呈剂量依赖性上调。间接免疫荧光染色结果也显示,高钙组Vimentin和FSP1荧光强度比低钙组显著增强。结论:钙离子浓度的变化能诱导人腹膜间皮细胞E-cadherin、FSP1、Vimentin蛋白表达发生变化,提示钙离子可能是参与调控间皮细胞间充质转分化的一个重要因素。
