Li Fang, Wang Yu, Tian Jianwei, Zhou Zhanmei, Yin Wei, Qin Xianhui, Wang Huizhen, Zeng Tao, Li Aiqing, Jiang Jianping
Division of Nephrology, Nanfang Hospital, Southern Medical University, National Clinical Research Center for Kidney Disease, State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Renal Failure Research and Guangdong Provincial Clinical Research Center for Kidney Disease, Guangzhou, China.
Laboratory Animal Research Center of Nanfang Hospital, Southern Medical University, Guangzhou, China.
Front Pharmacol. 2022 Dec 1;13:962770. doi: 10.3389/fphar.2022.962770. eCollection 2022.
Peritoneal dialysis is a common renal replacement method for end-stage renal disease. Long-term peritoneal dialysis leads to peritoneal dialysis-related peritoneal fibrosis, which leads to a cessation of treatment. Calpain is a protein belonging to calcium-dependent endopeptidase family and plays an important role in extracellular matrix remodeling. Here, we evaluated the effect of calpain in peritoneal dialysis-related peritoneal fibrosis. We established two animal models of peritoneal fibrosis and inhibited the activity of Calpain, and then collected peritoneal tissue to evaluate the progress of fibrosis and the changes of Calpain and β-catenin. We obtained Rat peritoneal mesothelial cells and Human peritoneal mesothelial cell line and stimulated with TGF-β to produce extracellular matrix. Next we inhibited Calpain activity or reduced Calpain9 expression, and then assessed changes in extracellular matrix and β-catenin. Inhibition of calpain activity attenuated chlorhexidine glucose and peritoneal dialysis-induced peritoneal thickening and β-catenin expression in mice. In addition, compared with the control group, when primary rat peritoneal mesothelial cells or human peritoneal mesothelial cells were treated with transforming growth factor beta, down-regulation of calpain activity inhibited the expression of Fibronectin and Collagen I, and increased the expression of E-cadherin. These changes could be adjusted after silencing calpain9. Finally, calpain9 deficiency was associated with down-regulation of Fibronectin and β-catenin in human peritoneal mesothelial cells. Our results suggest that calpain9 may be a key molecule in mediating peritoneal dialysis-related peritoneal fibrosis.
腹膜透析是终末期肾病常见的肾脏替代治疗方法。长期腹膜透析会导致腹膜透析相关的腹膜纤维化，进而导致治疗中断。钙蛋白酶是一种属于钙依赖性内肽酶家族的蛋白质，在细胞外基质重塑中起重要作用。在此，我们评估了钙蛋白酶在腹膜透析相关腹膜纤维化中的作用。我们建立了两种腹膜纤维化动物模型并抑制钙蛋白酶的活性，然后收集腹膜组织以评估纤维化进展以及钙蛋白酶和β-连环蛋白的变化。我们获取了大鼠腹膜间皮细胞和人腹膜间皮细胞系，并用转化生长因子-β刺激以产生细胞外基质。接下来我们抑制钙蛋白酶活性或降低钙蛋白酶9的表达，然后评估细胞外基质和β-连环蛋白的变化。抑制钙蛋白酶活性可减轻小鼠中葡萄糖酸氯己定和腹膜透析诱导的腹膜增厚以及β-连环蛋白表达。此外，与对照组相比，当用转化生长因子-β处理原代大鼠腹膜间皮细胞或人腹膜间皮细胞时，钙蛋白酶活性的下调抑制了纤连蛋白和I型胶原的表达，并增加了E-钙黏蛋白的表达。沉默钙蛋白酶9后这些变化可以得到调节。最后，钙蛋白酶9缺乏与人腹膜间皮细胞中纤连蛋白和β-连环蛋白的下调有关。我们的结果表明，钙蛋白酶9可能是介导腹膜透析相关腹膜纤维化的关键分子。
