Liu Yinghong, Dai Hongqin, Liu Fuyou, Sun Lin, Xiao Li, Liu Hong
Department of Nephropathy, Central South University, Changsha, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2012 Feb;37(2):190-6. doi: 10.3969/j.issn.1672-7347.2012.02.014.
To investigate the expression of galectin-1 with the stimulation of peritoneal dialysis solution (PDS) and its role in the epithelial-to-mesenchymal transition (EMT) in human peritoneal mesothelial cells (HPMCs).
HPMCs were stimulated with PDS containing different concentrations of high glucose (1.5%, 2.5%, and 4.25%). After 24 h, mRNA and protein expressions of galectin-1,vimentin, and zo-1 were analyzed with real-time PCR and Western blot, respectively. Liposome transfected siRNA technique was used to knock down the expression of galectin-1 and the effect of galectin-1 siRNA on the EMT of HPMCs was also observed under 4.25% PDS condition.
mRNA expression of galectin-1 in HPMCs increased in PDS groups, especially in group with 4.25% PDS (P<0.05). Protein expression of galectin-1 in HPMCs significantly increased in PDS groups with a dose dependent manner (P<0.05).Expression of vimentin in HPMCs significantly increased in PDS groups, especially in groups of 2.5% PDS and 4.25% PDS (P<0.05), but zo-1 expression markedly decreased (P<0.05). The expression of galectin-1 correlated positively with vimentin (P<0.05) but negatively with zo-1 (P<0.05). Expression of vimentin in groups of 4.25% PDS was markedly inhibited (P<0.05) by galectin-1 siRNA, whereas zo-1 expression was significantly increased (P<0.05).
Galectin-1 can mediate high glucose PDS-induced EMT in HPMCs and may be a new target for the prevention and treatment of peritoneal fibrosis.
研究半乳糖凝集素-1在腹膜透析液(PDS)刺激下的表达及其在人腹膜间皮细胞(HPMCs)上皮-间质转化(EMT)中的作用。
用含不同浓度高糖(1.5%、2.5%和4.25%)的PDS刺激HPMCs。24小时后,分别用实时PCR和蛋白质印迹法分析半乳糖凝集素-1、波形蛋白和紧密连接蛋白-1的mRNA和蛋白质表达。采用脂质体转染小干扰RNA(siRNA)技术敲低半乳糖凝集素-1的表达,并在4.25%PDS条件下观察半乳糖凝集素-1 siRNA对HPMCs EMT的影响。
PDS组HPMCs中半乳糖凝集素-1的mRNA表达增加,尤其是在4.25%PDS组(P<0.05)。PDS组HPMCs中半乳糖凝集素-1的蛋白质表达呈剂量依赖性显著增加(P<0.05)。PDS组HPMCs中波形蛋白的表达显著增加,尤其是在2.5%PDS和4.25%PDS组(P<0.05),但紧密连接蛋白-1的表达明显降低(P<0.05)。半乳糖凝集素-1的表达与波形蛋白呈正相关(P<0.05),与紧密连接蛋白-1呈负相关(P<0.05)。半乳糖凝集素-1 siRNA显著抑制4.25%PDS组波形蛋白的表达(P<0.05),而紧密连接蛋白-1的表达显著增加(P<0.05)。
半乳糖凝集素-1可介导高糖PDS诱导的HPMCs EMT,可能是腹膜纤维化防治的新靶点。