葛根素对体外培养人腹膜间皮细胞保护作用的研究

Puerarin protects the cultured human peritoneal mesothelial cells from the damage caused by peritoneal dialysate

目的:了解葛根素对体外培养的人腹膜间皮细胞增殖活性、氧化应激和转移生长因子-β1(TGF-1β)分泌的影响。方法:体外培养人腹膜间皮细胞,分对照组,腹膜透析液(PDS)组及葛根素1、2、3组(葛根素终浓度依次为50、100、200μg.m l-1)5组观察。检测各组间皮细胞的增殖活性,上清MDA、SOD、GSH水平以及TGF-β1的含量并进行比较。结果:在PDS干预下腹膜间皮细胞MTT还原能力明显下降,上清液MDA水平较对照组显著升高,SOD、GSH水平较对照组显著降低。葛根素组MDA水平显著低于PDS组,SOD、GSH水平显著高于PDS组。腹膜间皮细胞在PDS干预下分泌TGF-1β较对照组显著增加,葛根素组TGF-β1分泌与PDS组比较有显著下降。结论:葛根素可以拮抗商业性PDS对腹膜间皮细胞增殖活性的抑制作用,可以抑制氧化应激和TGF-β1分泌。

Objective To investigate the effect of Puerarin on the cell proliferation, oxidative stress and the secretion of TGF-β1 in cultured human peritoneal mesothelial cell（HPMC）. Methods HPMC cultured in vitro were divided into 5 groups, RPMI 1640 culture medium as control, peritoneal dialysis solution（PDS） containing 2.5% glucose as PDS group,PDS containing 2.5% glucose and 100,200 or 400μg·ml^-1 Puerarin respectively as Puerarin group 1, Puerarin group 2 and Puerarin group 3. Each group was supplemented with equal volum of RPMI 1640 culture medium containing 20% FBS. After 48 h incubating, mitochondrial dehydrogenases activity （ MTT assay）, the levels of TGF-β1, MDA, SOD and GSH in supernatant of the cell culture medium were detected and co chondrial dehydrogenases activity was observed in PDS groups compared with those in control group and Puerarin groups （P 〈0.01 ）. The level of MDA were much lower in control group and Puerarin groups than that in PDS group （P 〈 0.05 ）. And, higher level of SOD and GSH were observed in control group and Puerarin groups than those in PDS group （P 〈 0.05 ）. Significant lower level of TGF-β1, in supernatant was found in control group, as well as in Puerarin groups, when compared with that in PDS group（P 〈0.01 ）. Conclusion Puerarin can restore the cell proliferation activity reduced by peritoneal dialyste. Also, it reduces the oxidative stress and the secretion of TGF-β1, induced by commercial PDS in cultured human peritoneal mesothelial cells.