北五味子乙素抗AngⅡ诱导的心肌成纤维细胞增殖及作用机制

Antiproliferative Effect of Schisandrin B on Cardiac Fibroblast via Oxidation Mechanism in Rats

摘要:

目的：探讨北五味子木脂素（schisandra lignin，SCL）中的活性成分之一五味子乙素（schisandrin B，Sch B）对血管紧张素Ⅱ（angiotensinⅡ，AngⅡ）诱导心肌成纤维细胞（cardiac fibroblast，CFb）增殖的作用及作用机制。方法：以培养的新生Wistar大鼠乳鼠CFb为实验模型，实验分为对照组、AngⅡ模型（10^-1μmol·L^-1）组、Sch B 1，10，30 μmol·L^-1 3个剂量组。采用胰酶消化、差速贴壁法培养CFb，MTT法测定CFb增殖活力；分光光度计测定CFb中超氧化物歧化酶（SOD）活力和丙二醛（MDA）含量；流式细胞仪检测线粒体膜电位水平（mitochondrial membrane potential，MMP）ΔΨm；激光共聚焦显微镜检测细胞内钙离子水平。结果：AngⅡ组SOD 活力（122.72±21.59） U·mL^-1低于对照组（135.81±26.62）U·mL^-1，P<0.01，MDA水平（2.82±0.14） μmol·L^-1高于对照组（1.17±0.06） μmol·L^-1，P<0.01。SchB 1~30 μmol·L^-1能显著提高CFb中SOD活力，降低MDA含量，与AngⅡ相比（P<0.05或P<0.01）；AngⅡ组线粒体膜电位明显下降（587.6±55.2）ΔΨm，钙离子水平升高（335.6±28.3），与对照组比较（3 069.2±282.4）ΔΨm，（50.7±4.6），P<0.01或P<0.001，给予Sch B处理后ΔΨm升高（Sch B 10，30 μmol·L^-1），钙离子水平下降（SchB 1~30 μmol·L^-1），与AngⅡ组比较（P<0.05或P<0.01）。结论：Sch B通过提高CFb SOD活力、降低MDA水平、增加清除自由基能力抑制CFb增殖，其作用与其升高线性体膜电位水平，抑制细胞内钙离子浓度，减少促进CFb增殖的氧化物质生成有关。

Abstract:

Objective: To probe the oxidative mechanism of schisandrin B (Sch B) on antiproliferation of neonatal rat cardiac fibroblast induced by AngII. Method: Cultured neonatal cardiac fibroblast (CFb) cells were randomly divided into control, AngiotensinⅡ (AngⅡ), 10^-1μmol·L^-1) and three doses of Sch B groups (SchB 1, 10, 30 μmol·L^-1).Trypsin digestion and spectrophotometer method were used to measure superoxide dismutase (SOD) activity and malondialdehyde (MDA) level. Flow cytometry was adopted to test mitochondrial membrane potential level. Laser confocal microscope technology was applied to measure intracellular calcium level. Result: SOD activity and MDA level in AngII group was lower and higher than that of control respectively (P<0.01), Sch B (1-30 μmol·L^-1) elevated SOD activity and drop MDA level of CFb (P<0.05 or P<0.01) respectively.mitochondrial membrane potential (MMP, ΔΨm) was increased (Sch B 10, 30 μmol·L^-1) and calcium intensity was declined (1, 30 μmol·L^-1) after Sch B treatment and have dramatic difference compared with that in AngⅡgroup. Conclusion: Sch B can elevate the free radical scavenging ability via enhancing SOD activity and decrease MDA level in CFb, these effects are related to increasing MMP levels and inhibiting the intracellular calcium concentration, therefore suppressing the generation of oxidized substances related to CFb proliferation.