丹参酮ⅡA对腹膜透析液诱导的氧化应激及腹膜间皮细胞凋亡因子表达的影响-东南大学学报医学版

丹参酮ⅡA对腹膜透析液诱导的氧化应激及腹膜间皮细胞凋亡因子表达的影响

单位：1. 南京医科大学鼓楼临床医学院, 江苏南京 210008;
2. 南京大学医学院附属南京鼓楼医院肾内科, 江苏南京 210008

分类号：R-33;R329.25

出版年·卷·期（页码）：2013·32·第一期（36-41）

摘要：

目的:探讨丹参酮ⅡA(TSN)注射液对高糖腹膜透析液(PDF)诱导体外培养人腹膜间皮细胞(HPMCs)氧化应激及凋亡的影响。方法:氧化应激指标检测:体外培养人网膜间皮细胞,分为对照组(DEME培养基)、PDF组(4.25%PDF)、TSN组(含TSN终浓度为100 μmol·L^-1的DEME培养基)、TSN低浓度组(含TSN终浓度为50 μmol·L^-1的4.25%PDF)、TSN高浓度组(含TSN终浓度为100 μmol·L^-1的4.25%PDF)5组观察,共培养48 h后取上清液检测丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)浓度。细胞凋亡因子检测:体外培养细胞分为对照组(DEME培养基)、H₂O₂组(3%H₂O₂)、TSN+H₂O₂组(含TSN终浓度为50 μmol·L^-1的3%H₂O₂)、PDF组(4.25%PDF)、TSN+PDF组(含TSN终浓度为50 μmol·L^-1的4.25%PDF)5组观察,共培养72 h后Real time PCR法检测细胞凋亡因子bax、caspase-3及抑制凋亡因子bcl-2的表达情况,Western Blot法检测细胞凋亡蛋白bax、caspase-3及抑制凋亡蛋白bcl-2的表达情况。结果:与对照组比较,PDF组中MDA的含量显著升高(P<0.01),SOD、GSH的含量显著降低(P<0.01)。与PDF组比较,添加TSN后的各组MDA含量明显降低(P<0.01),SOD、GSH的含量显著升高(P<0.01)。Real time PCR及Western Blot结果发现,与对照组比较,PDF组及H₂O₂组bax、caspase-3的mRNA及蛋白表达显著增多(P<0.01),同时bcl-2的mRNA及蛋白的表达显著减少(P<0.01);添加TSN的PDF组及H₂O₂组分别较单纯PDF组及H₂O₂组bax、caspase-3的mRNA及蛋白表达显著减少(P<0.01),而bcl-2的mRNA及蛋白表达显著增多(P<0.01)。结论:TSN可能通过抑制氧化应激下调PDF诱导的HPMCs细胞凋亡因子高表达,具有潜在的腹膜保护作用。

Objective: To investigate the effect of tanshinoneⅡA(TSN) on oxidative stress and apoptosis of human peritoneal mesothelial cells (HPMCs) stimulated by peritoneal dialysis fluid (PDF). Methods: Oxidative stress index detection: HPMCs cultured in vitro were divided into control group (DMEM medium), PDF group (4.25%PDF), TSN group (DMEM medium with 100 μmol·L^-1 TSN), TSN low concentration group (4.25%PDF with 50 μmol·L^-1 TSN), TSN high concentration group (4.25%PDF with 100 μmol·L^-1 TSN). After 48 hours the levels of MDA, SOD and GSH in the supernatant were detected. Apoptosis cytokines detection:HPMCs cultured in vitro were divided into control group (DMEM medium), H₂O₂ group (3%H₂O₂), TSN+H₂O₂ group (3%H₂O₂ with 50 μmol·L^-1TSN), PDF group (4.25%PDF), TSN+PDF (4.25%PDF with 50 μmol·L^-1TSN). After 72 hours of treatment, apoptosis cytokines bax, caspase-3 and inhibition apoptosis cytokines bcl-2 mRNA and protein were detected by Real time PCR and Western Blot, respectively.Results: Compared with control group, PDF group significantly increased the level of MDA (P<0.01), and decreased the level of SOD and GSH (P<0.01). Compared with PDF group, groups with TSN significantly decreased the level of MDA (P<0.01) and increased the levels of SOD and GSH (P<0.01). Compared with control group, PDF and H₂O₂ groups significantly increased the expressions of both bax and caspase-3 mRNA and protein (P<0.01), and decreased the expressions of bcl-2 mRNA and protein (P<0.01). Compared with PDF and H₂O₂ groups, PDF and H₂O₂ with TSN significantly decreased the expressions of both bax and caspase-3 mRNA and protein (P<0.01) and increased the expressions of bcl-2 mRNA and protein (P<0.01). Conclusion: TSN may down-regulate PDF induced high expression of apoptosis cytokines through inhibiting oxidative stress of HPMCs, and thus has a protective effect on peritoneal membrane.

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