黄芪注射液拮抗腹膜透析液诱导腹膜间皮细胞凋亡的研究

Effects of Astragalus on the apoptosis of cultured human peritoneal mesothelium induced by peritoneal dialysis solution

目的:了解黄芪注射液对体外培养的人腹膜间皮细胞凋亡的影响。方法:培养液中添加含不同浓度黄芪注射液的1.5%葡萄糖腹膜透析液(PDS),观察细胞caspase-3活性变化,应用Annexin V/PI双染色流式细胞仪方法以及末端标记技术(TUNEL)观察各组腹膜间皮细胞凋亡的情况。结果:以对照组caspase-3活性为1,PDS组细胞caspase-3相对活性为3.26±0.91,显著高于黄芪一组(1.87±0.43)和黄芪二组(1.67±0.32),P<0.05。An-nexin V-FITC/PI双标记结果显示,PDS组细胞凋亡率和死亡率分别为(46.8±14.2)%和(25.7±12.4)%,显著高于对照组的(9.04±4.86)%和(7.96±4.28)%,P<0.05;黄芪一组和黄芪二组的细胞凋亡率和死亡率显著低于PDS组(P<0.05)。TUNEL研究结果显示,PDS组细胞凋亡明显高于对照组、黄芪一组和黄芪二组。结论:黄芪注射液可抑制PDS诱导的腹膜间皮细胞凋亡的发生。

Objective ： To investigate the effects of Astragalus on the the apoptosis of cultured human peritoneal mesothelium induced by peritoneal dialysis solution （PDS）. Methods： Using RPM1640 culture medium as control , PDS group（ commercial PDS containing 1.5% glucose without Astragalus）, Astragalus group 1 （commercial PDS containing 1.5% glucose and 1% Astragalus） and Astragalus group 2 （commercial PDS containing 1.5% glucose and 2% Astragalus） were tested to explore the apoptosis of cultured human peritoneal mesothelial cell in each group. The activity of caspase-3, the terminal deoxynucleotidyl transferase （TdT） mediated dUTP nick end labeling （TUNEL） and flow cytometry analysis were used in apopto- sis analysis in this study. Results ： The activity of caspase-3 in the control group was defined as 1, the relative activity of caspase-3 in PDS group, Astragalus group 1 and Astragalus group 2 were 3.26 ±0.91, 1.87±0.43 and 1.67 ±0.32 respectively. The activity of caspase-3 in PDS group was significantly higher than those in the Astragalus group 1 and Astragalus group 2 （P 〈 0.05 ）. Necrosis rate of mesothelial cells detected by uptake of propidium iodide （ FACS ） and apoptosis rate by annexin V in PDS group were （25.7 ± 12.4 ）% and （46.8 ± 14.2 ）% respectively. Although markedly higher rate could be found in Astragalus group 1 and Astragalus group 2 as copmared with those in the control group, significantly lower rate of necrosis and apoptosis could be found in Astragalus group 1 and Astragalus group 2 as cpmared with those in PDS group（P 〈0.05）. Apoptosis rate detected by the method of TUNNEL in PDS group were much higher than those in the control, Astragalus group 1 and Astragalus group 2. Conclusion ： Astragalus can reduce the apoptosis of cultured human peritoneal mesothelium induced by PDS in vitro.