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  

Effects of tanshinoneⅡA on oxidative stress and apoptosis cytokine expression of human peritoneal mesothelial cells

作　　者：夏阳阳[1,2] 蒋春明[2] 孙琤[2] 于立杰[2] 张苗[1,2] 

机构地区：[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

出　　处：《东南大学学报（医学版）》2013年第1期36-41,共6页Journal of Southeast University(Medical Science Edition)

基　　金：南京市重点科技发展项目(ZKX08026)

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100μmol.L-1的DEME培养基)、TSN低浓度组(含TSN终浓度为50μmol.L-1的4.25%PDF)、TSN高浓度组(含TSN终浓度为100μmol.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50μmol.L-1的3%H2O2)、PDF组(4.25%PDF)、TSN+PDF组(含TSN终浓度为50μmol.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 tanshinone Ⅱ A （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 μ mol· L-1 TSN） , TSN low concentration group （4. 25% PDF with 50μ mol · 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）, H202 group （3%H202 ） , TSN ＋ H202 group （ 3 % H202 with 50 μmol · L - 1TSN ）, PDF group （4. 25 % PDF）, TSN ＋ PDF （4. 25 % PDF with 50μmol· L- TSN）. 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 H202 groups significantly increased the expressions of both bax and caspase- 3 mRNA and protein （P 〈 0. 01 ） , and decreasedthe expressions of bcl-2 mRNA and protein （ P 〈 0.01 ） with TSN significantly decreased the expressions of both increased the expressions of bcl-2 mRNA and protein （ induced high expression of apoptosis cytokines protective effect on peritoneal membrane. through Compared with PDF and bax and caspase-3 mRNA P 〈 0. 01 ）. Conclusion： inhibiting oxidative stress H202 groups, PDF and H202 and protein （ P 〈 0. 01 ） and TSN may down-regul

关 键 词：丹参酮ⅡA 腹膜透析液 氧化应激 凋亡因子 抑制凋亡因子 

分 类 号：R-33[医药卫生] R329.25