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目的建立体外血脑屏障(BBB)模型,考察血栓通的主要活性成分(人参皂苷Rb1、人参皂苷Rd、人参皂苷Re、人参皂苷Rg1及三七皂苷R1)的通透性。方法选用小鼠脑微血管内皮细胞株bEnd.3建立体外BBB模型。然后分为正常组和模型组,正常组培养6 h,模型组缺氧6 h后,均给予50μg·m L~(-1)血栓通0.2 m L。给药4 h后,用高效液相色谱/质谱联用法测定各组的各有效成分的通透系数。结果正常组中,人参皂苷Rg1、人参皂苷Rb1、人参皂苷Rd、人参皂苷Re、三七皂苷R1的通透系数分别为(70.37±5.78)×10~(-8),(102.36±69.21)×10~(-9),(0.57±0.38)×10~(-9),(98.96±15.52)×10~(-9),(382.18±42.49)×10~(-9)cm·s~(-1);模型组中,这5种活性成分的通透系数分别为(116.20±21.80)×10~(-8),(555.40±202.65)×10~(-9),(2.96±1.43)×10~(-9),(202.66±37.5)×10~(-9),(634.26±124.40)×10~(-9)cm·s~(-1)。与正常组相比,模型组的通透系数显著增加,差异有统计学意义(P<0.05)。结论血脑屏障缺氧6 h后明显开放,注射用血栓通(冻干)可以透过血脑屏障发挥神经保护作用。
OBJECTIVE To establish an in vitro model of blood-brain barrier (BBB) and investigate the permeability of the main active components of Xueshuantong (ginsenoside Rb1, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1 and notoginsenoside R1). Methods Mouse brain microvascular endothelial cell line bEnd.3 selected in vitro BBB model. Then, the rats were divided into normal group and model group. Normal group were cultured for 6 h. After hypoxia for 6 h in model group, 50 μg · m L -1 of thrombus was given 0.2 m L. After 4 h of administration, the permeation coefficient of each active ingredient in each group was determined by high performance liquid chromatography / mass spectrometry. Results In the normal group, the permeability coefficients of ginsenoside Rg1, ginsenoside Rb1, ginsenoside Rd, ginsenoside Re and notoginsenoside R1 were (70.37 ± 5.78) × 10 -8 and (102.36 ± 69.21) × (9), (0.57 ± 0.38) × 10 ~ (-9), (98.96 ± 15.52) × 10 ~ (-9), (382.18 ± 42.49) × 10 ~ (-9) cm · s ~ -1). In the model group, the permeability coefficients of the five active ingredients were (116.20 ± 21.80) × 10 -8 (555.40 ± 202.65) × 10 -9 (2.96 ± 1.43) × 10 ~ (-9), (202.66 ± 37.5) × 10 ~ (-9), (634.26 ± 124.40) × 10 ~ (-9) cm · s ~ (-1). Compared with the normal group, the model group of permeability coefficient increased significantly, the difference was statistically significant (P <0.05). Conclusion The blood-brain barrier is obviously open after 6 h of hypoxia. Xueshuantong injection (lyophilization) can play a neuroprotective role through the blood-brain barrier.