目的 研究丹参川芎有效成分不同配伍对缺氧损伤大鼠脑微血管内皮细胞（rat brain microvascular endothelial cells, rBMECs）的影响，并找出较优组合。方法 采用体外培养的rBMECs，并进行兔抗鼠Ⅷ因子鉴定，建立体外细胞缺氧模型。对参芎有效成分丹参素（A）、丹参酮Ⅱa（B）、川芎嗪（C）、藁本内酯（D）、阿魏酸（E），以3个剂量水平进行正交实验，另设正常组、模型组，观察不同配伍对细胞中超氧化物歧化酶（superoxide dismutase, SOD）活性、细胞上清液乳酸脱氢酶（lactate dehydrogenase, LDH）、细胞凋亡率及细胞周期的影响。采用正交方差分析、直观综合分析对实验结果进行多指标优化。 结果 与模型组比较，各配伍样品均可提高SOD活性，降低LDH含量，减少细胞早期凋亡，抑制缺氧诱导的rBMECs发生G1/S期阻滞的作用。正交方差分析表明：A2B1C3D2E1为SOD的较优组合，A3B1C1D2E3为LDH的较优组合，A2B1C2D3E2为早期凋亡的较优组合，A2B3C3D3E2为G1期的较优组合，A2B3C3D2E2为S期的较优组合；综合分析结果显示A2B1C1D3E2为多指标的较优组合。结论 丹参川芎有效成分不同配伍可在一定程度上保护缺氧对rBMEC的损伤，不同配伍作用方式的侧重不同。
Objective To investigate the effects of different doses of active ingredients in Salvia miltiorrhiza and Rhizoma Ligustici Chuanxiong on rat brain microvascular endothelial cells (rBMECs) during hypoxic injury and to determine the preferred dose combinations. Methods rBMECs were cultured in vitro and were identified with rabbit-anti-rat factor Ⅷ antibody. The rBMECs were divided into normal group, model group, and test groups. An in vitro cell model of hypoxia was induced in the model group and test groups. The test groups were treated with different doses of tanshinol (A), tanshinone ⅡA (B), ligustrazine (C), ligustilide (D), and ferulic acid (E). An orthogonal test was performed to investigate the effects of these active ingredients in Salvia miltiorrhiza and Rhizoma Ligustici Chuanxiong (each at three dose levels) on superoxide dismutase (SOD) activity in cells, lactate dehydrogenase (LDH) content in cell supernatant, cell apoptosis rate, and cell cycle. Orthogonal analysis of variance and direct comprehensive analysis were used to optimize the dose combination for these indices. Results Compared with the model group, the test groups had increased SOD activity, decreased LDH content, reduced early apoptosis, and inhibited G1/S arrest induced by hypoxia. The orthogonal analysis of variance showed that A2B1C3D2E1 was the preferred dose combination for SOD, A3B1C1D2E3 was the preferred dose combination for LDH, A2B1C2D3E2 was the preferred dose combination for early apoptosis, A2B3C3D3E2 was the preferred dose combination for G1 phase, and A2B3C3D2E2 was the preferred dose combination for S phase. The comprehensive analysis showed that A2B1C1D3E2 was the preferred dose for multiple indices. Conclusion Different doses of active ingredients in Salvia miltiorrhiza and Rhizoma Ligustici Chuanxiong can reduce the hypoxic injury of rBMECs, and their action mechanism varies in different dose combinations.
管敏国,张宇燕,万海同,周 鹏,杨 珍,杨洁红.丹参川芎有效成分配伍对缺氧损伤大鼠脑内皮细胞的影响[J].安徽中医药大学学报,2014,33(2):74-78复制