新多聚糖碳酸氢盐腹膜透析液对人腹膜间皮细胞凋亡及纤维化相关因子的影响

The effect of the new polysaccharide bicarbonate peritoneal dialysis fluid on apoptosis and fibrosis of peritoneal mesothelial cells

目的:比较新研制的多聚糖碳酸氢盐腹膜透析液(PDF)与常用乳酸盐PDF对人腹膜间皮细胞(HPMC)凋亡及间质纤维化相关因子的影响。方法:体外培养HPMC细胞,使用新型PDF与不同葡萄糖浓度的传统乳酸盐PDF(1.5%、2.5%、4.25%)处理细胞,通过实时荧光定量PCR法检测细胞凋亡因子Bax、胱天蛋白酶3(Caspase-3)及Bcl-2、转化生长因子β1(TGF-β1)、纤维连接蛋白(FN)的mRNA表达量,ELISA法测定细胞分泌TGF-β1量,免疫印迹法检测细胞凋亡蛋白Bax、Caspase-3、抑制凋亡蛋白Bcl-2和FN的表达量。结果:不同PDF刺激HPMC细胞一定时间后,1.5%、2.5%、4.25%传统PDF中葡萄糖浓度越高,Bax、Caspase-3、TGF-β1和FN增高越明显,抑制凋亡因子Bcl-2下降越显著,而新型PDF影响较小。含2.5%、4.25%葡萄糖的PDF刺激后,纤维化相关因子TGF-β1、FN明显高于阴性对照组和新型PDF组(均P<0.05)。结论:新型多聚糖碳酸氢盐PDF对腹膜间皮细胞凋亡信号活化低于传统乳酸盐PDF,其腹膜间皮细胞纤维化程度低,有很好的临床应用前景。

Objective： To compare the influence of new polysaccharide bicarbonate peritoneal dialysis fluids（ PDF） on the human peritoneal mesothelial cells（ HPMC） apoptosis and interstitial fibrosis factors. Methodology：Human peritoneal mesothelial cells（ HPMC） were cultured in vitro,then treated with the new PDF and traditional PDF with different concentration of glucose（ 1. 5%,2. 5%,4. 25%）. Real-time quantitative PCR was used to detect the mRNA expression of apoptotic factors Bax,Caspase-3 and apoptosis inhibition factor Bcl-2,TGF-β1 and FN. ELISA determined the secretion of TGF-β1 expression,western blot was used to detect apoptosis protein Bax,Caspase-3,apoptosis inhibition proteins Bcl-2 and FN. Results： After a certain time of stimulation on HPMC,the higher the glucose concentration（ 1. 5% 、2. 5% 、4. 25%） of traditional PDF,Bax,Caspase-3,TGF-β1,FN increased more significantly,apoptosis inhibition factor Bcl-2 decreased more significantly. After stimulation with 2. 5%,4. 25% PDF,fibrosis related factor TGF-β1 and FN were much higher than that of the negative control group and new PDF group（ P〈0. 05）. Conclusion： The new polysaccharide bicarbonate peritoneal dialysis fluid is superior to the traditional peritoneal dialysis fluid,its effect on the apoptosis protein of peritoneal mesothelial cells is weaker than that of the traditional peritoneal dialysis fluid,peritoneal mesothelial cell fibrosis degree is aslo lower,which has good prospects for clinical application.