AIM To elucidate the mechanism of actions of salvianolic acids on cardiovascular system. METHODS The viability of cultured human umbilical vein endothelial cells(HUVEC) was measured by MTT assay. Apoptosis was assessed by propidium iodide (PI) staining and TUNEL assay. Caspase-3 activity and [Ca2+]i in HUVEC were measured by fluorescent assays. RESULTSThe results showed that the viability of HUVEC treated with 100 μmol*L-1 H2O2 for 8 h decreased by 40.2%. Salvianolic acids 10 and 100 mg*L-1 increased the viability of H2O2-treated cells by 8.4% and 28.6%, respectively. The apoptotic rate of HUVEC tested by PI staining increased from (7.1±0.7)% to (38.1±4.0)%after 18 h of treatment with H2O2. Salvianolic acids 100 mg*L-1 reduced the apoptotic rate to (13.6±0.8)%. Under the same condition tested by TUNEL assay, the number of TUNEL-positive cells increased from (3.0±0.8)% to (30.5±2.9)%. Salvianolic acids 10 and 100 mg*L-1 reduced the number to (21.5±2.4)% and (19.0±3.7)%, respectively. In addition, H2O2 increased caspase-3 activity in HUVEC by 176% and the [Ca2+]i by 101%. Salvianolic acids inhibited elevation of caspase-3 activity and [Ca2+]i induced by H2O2. CONCLUSION Salvianolic acids have protective effect on endothelial cells against damage induced by H2O2.%目的研究总丹酚酸对过氧化氢(H2O2)诱导血管内皮细胞损伤的影响.方法培养的人脐静脉内皮细胞,用MTT法测定内皮细胞的存活率.用碘化丙啶(PI)染色和TUNEL法测定H2O2诱导内皮细胞的凋亡作用.用荧光法测定半胱天冬酶(caspase)-3的活性.用Fura-2/AM测定内皮细胞内游离钙离子浓度([Ca2+]i).结果 100 μmol*L-1 H2O2可使内皮细胞的存活率下降40.2%,提前1 h给予总丹酚酸10 和100 mg*L-1使存活率分别增加8.4%和28.6%.100 μmol*L-1 H2O2还可引起内皮细胞的凋亡,PI染色的结果显示,H2O2处理内皮细胞18 h后,凋亡率从(7.1%±0.7)%升至(38.1±4.0)%.总丹酚酸100 mg*L-1提前处理内皮细胞1 h,可使内皮细胞凋亡率降至(13.6±0.8)%.TUNEL方法检测结果显示,H2O2处理内皮细胞后,TUNEL染色阳性的细胞从(3.0±0.8)%升至(30.5±2.9)%,总丹酚酸100 mg*L-1可使TUNEL阳性细胞率降至(19.0±3.7)%.此外,H2O2处理细胞后,内皮细胞半胱天冬酶-3的活性增加176%, [Ca2+]i升高101%,而总丹酚酸可以抑制H2O2引起的半胱天冬酶-3活性的增加和[Ca2+]i升高.结论总丹酚酸对H2O2引起的内皮细胞损伤具有明显的抑制作用.
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