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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 1,10,30μmol·L-13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 1~30μmol·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 10,30μmol·L-1),钙离子水平下降(SchB 1~30μmol·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。
OBJECTIVE: To investigate the effect of Schisandrin B (Sch B), one of the active ingredients in Schisandra lignin (SCL) on angiotensin Ⅱ (AngⅡ) -induced cardiac fibroblast (CFb) ) Proliferation and its mechanism of action. Methods: CFb from neonatal Wistar rats were used as experimental model. The experimental group was divided into control group, Ang Ⅱ model group (10-1μmol·L-1), Sch B 1,10,30μmol·L-13 dose group. CFb was cultured with trypsin digestion and differential adhesion method, the activity of CFb proliferation was measured by MTT method, the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in CFb were measured by spectrophotometer, the mitochondrial membrane The level of mitochondrial membrane potential (MMP) ΔΨm was measured by laser confocal microscopy. Results: Compared with control group, the activity of SOD (122.72 ± 21.59) U · mL-1 in AngⅡ group was significantly lower than that in control group (135.81 ± 26.62) U · mL-1, P <0.01 and MDA level (2.82 ± 0.14) μmol·L- Group (1.17 ± 0.06) μmol·L-1, P <0.01. Compared with AngⅡ (P <0.05 or P <0.01), SchB 1 ~ 30μmol·L-1 could significantly increase SOD activity and decrease MDA content in CFb (587.6 ± 55.2) ΔΨm, (ΨΨm, (50.7 ± 4.6), P <0.01 or P <0.001, respectively) compared with that of the control group (Sch B 10, 30 μmol · L-1), and the level of calcium decreased (SchB 1 ~ 30μmol·L-1) compared with AngⅡ group (P <0.05 or P <0.01). CONCLUSIONS: Sch B is related to increasing CFb SOD activity, decreasing MDA level and increasing free radical scavenging ability of CFb, which is related to increasing the level of linear membrane potential, inhibiting the intracellular calcium concentration and decreasing the production of oxidants that promote CFb proliferation .