论文部分内容阅读
目的:研究丹酚酸B镁盐对低氧引起的内皮细胞内钙升高和一氧化氮释放增加的抑制作用。方法:培养的人脐静脉内皮细胞暴露在95％N_2+5％CO_2条件下30分钟。内皮细胞的损伤用染料排除实验和LDH的释放来评价。胞内游离钙浓度用钙荧光探针Fura2-AM测定。一氧化氮含量用一氧化氮试剂盒测定。内皮型一氧化氮合酶(eNOS)mRNA和诱导型一氧化氮合酶(iNOS)mRNA的表达用半定量逆转录聚合酶链式反应检测。结果:低氧引起内皮细胞的活力由正常条件下(93.0±2.6)％降至(85.5±2.1)％(P<0.01),并导致内皮细胞LDH的释放由(41±28)U/L增加至(141±68)U/L(P<0.01)。丹酚酸B镁盐5mg/L和10mg/L能明显提高内皮细胞活力和降低LDH释放量。同时,低氧还显著增加内皮细胞的胞内游离钙浓度和一氧化氮释放,并上调其eNOS mRNA和iNOS mRNA的表达(P<0.01)。丹酚酸B镁盐5mg/L和10mg/L能抑制其增加(P<0.01),但在无钙的条件下,低氧对其均没有影响。结论:丹酚酸B镁盐能改善低氧引起的内皮细胞损伤,抑制低氧引起的内皮细胞内钙浓度的升高,一氧化氮释放,eNOS和iNOS mRNA表达的增加。丹酚酸B镁盐对一氧化氮产生和eNOSmRNA表达的抑制可能与其抑制低氧引起内皮细胞的胞外钙内流有关。
Objective: To study the inhibitory effect of salvianolic acid B magnesium salt on hypoxia-induced increase of intracellular calcium and nitric oxide release. Methods: Cultured human umbilical vein endothelial cells were exposed to 95% N 2 + 5% CO 2 for 30 minutes. Endothelial cell damage was assessed using dye exclusion experiments and LDH release. The intracellular free calcium concentration was measured with the calcium fluorescent probe Fura2-AM. Nitric oxide levels were measured using a nitric oxide kit. The expression of endothelial nitric oxide synthase (eNOS) mRNA and inducible nitric oxide synthase (iNOS) mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction. Results: The hypoxia induced a decrease in endothelial cell viability from (93.0 ± 2.6)% to (85.5 ± 2.1)% (P <0.01) and an increase in endothelial cell LDH release from (41 ± 28) U / L To (141 ± 68) U / L (P <0.01). Salvianolic acid B magnesium salt 5mg / L and 10mg / L can significantly improve endothelial cell viability and reduce LDH release. Hypoxia also significantly increased the intracellular free calcium concentration and nitric oxide release and up-regulated the expression of eNOS mRNA and iNOS mRNA in endothelial cells (P <0.01). Salvianolic acid B magnesium 5 mg / L and 10 mg / L inhibited the increase (P <0.01), but in the absence of calcium, hypoxia had no effect on them. CONCLUSION: Salvianolic acid B magnesium salt can improve endothelial cell injury induced by hypoxia, inhibit the increase of intracellular calcium concentration induced by hypoxia, the release of nitric oxide and the expression of eNOS and iNOS mRNA. Inhibition of salvianolic acid B magnesium salt on nitric oxide production and eNOS mRNA expression may be related to its inhibition of hypoxia-induced extracellular calcium influx in endothelial cells.