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目的:比较补阳还五汤含药血清和含药血浆对缺氧培养的大鼠脑微血管内皮细胞(BMECs)纤溶系统和黏附分子的表达与体内实验真实值的接近程度,为半体内实验选择含药血浆还是含药血清提供参考依据。方法:用颈内动脉线栓法建立局灶性脑缺血模型,缺氧培养箱孵育BMECs,采用ELISA法测定PAI-1、t PA的表达,RT-PCR法检测ICAM-1、VCAM-1 mRNA的含量,Western Blot法测定ICAM-1、VCAM-1蛋白的含量。结果:补阳还五汤含药血浆较含药血清更显著降低缺氧损伤大鼠BMECs PAI-1、增强t PA水平,含药血浆和含药血清均能明显降低ICAM-1、VCAM-1、ICAM-1 mRNA及VCAM-1 mRNA含量,与空白血浆、空白血清比较,差异显著(P<0.05);半体内实验的含药血浆与体内实验真实值各指标相关系数均高于含药血清。结论:含药血浆比含药血清更接近体内真实情况,在进行半体内实验研究中药复方对纤溶系统和黏附分子蛋白及mRNA表达的影响时,特别是研究纤溶系统PAI-1和t PA指标的表达时,建议优先选择血浆药理学方法。
OBJECTIVE: To compare the close relationship between the expression of fibrinolytic system and adhesion molecules in rat brain microvascular endothelial cells (BMECs) cultured in hypoxia and the real experimental values of Buyang Huanwu Decoction-containing serum and drug-containing plasma for in-vivo experiments Select drug-containing plasma or drug-containing serum provide a reference. Methods: The model of focal cerebral ischemia was established by the method of internal carotid artery occlusion. The expression of PAI-1 and t PA was detected by ELISA and ICAM-1 and VCAM-1 by RT-PCR mRNA content, Western Blot determination of ICAM-1, VCAM-1 protein content. Results: The drug-containing plasma of Buyang Huanwu Decoction could significantly decrease PAI-1 and PAI-1 of BMECs in hypoxia-injured rats compared with the control group, and both plasma and drug-containing serum could significantly decrease ICAM-1 and VCAM-1 , ICAM-1 mRNA and VCAM-1 mRNA were significantly different from those in blank plasma and blank serum (P <0.05). The correlation coefficients of the real plasma values of in-vivo experiments and in vivo experiments were higher . CONCLUSION: The drug-containing plasma is closer to the real situation in vivo than the drug-containing serum. In the study of the influence of Chinese herbal compound on the fibrinolytic system and the expression of adhesion molecule protein and mRNA in a semi-vivo experiment, especially the fibrinolytic system PAI-1 and t PA Indicators of the expression, it is recommended to give priority to the method of plasma pharmacology.