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目的:探讨川芎嗪与葛根素合用对糖氧剥夺诱导的体外血脑屏障(aBBB)通透性的影响。方法:实验分为正常对照组、模型组、川芎嗪组、葛根素组、葛根素与川芎嗪合用组和尼莫地平组6个组。原代培养大鼠脑微血管内皮细胞和星形胶质细胞共培养构建aBBB模型,经糖氧剥夺诱导处理建立aBBB损伤模型,跨内皮电阻抗、γ-谷氨酰转肽酶、碱性磷酸酶及乳酸脱氢酶鉴定aBBB损伤情况,高效液相色谱法测定川芎嗪、葛根素及尼莫地平的含量,并计算在OGD诱导下的aBBB药物透过浓度。结果:与对照组相比,模型组TEER值降低(P<0.01),川芎嗪组、川芎嗪与葛根素合用组、尼莫地平组-γGT,AKP升高(P<0.01),与川芎嗪与葛根素合用组相比,川芎嗪、葛根素单用AKP升高明显(P<0.01),尼莫地平各指标均有显著性差异(P<0.05),提示川芎嗪与葛根素合用在可升高TEER,增加-γGT,AKP含量和降低LDH具有协同作用,OGD诱导下aBBB模型的培液中川芎嗪与葛根素合用组各成分透过浓度均较川芎嗪、葛根素单用增加。结论:川芎嗪与葛根素合用较两者单用易透过aBBB,且对aBBB有一定的保护作用,可能与降低OGD诱导下的aBBB通透性有关。
Objective: To investigate the effect of ligustrazine and puerarin on permeability of blood-brain barrier (aBBB) induced by glucose-oxygen deprivation. Methods: The experiment was divided into 6 groups: normal control group, model group, ligustrazine group, puerarin group, puerarin and ligustrazine group and nimodipine group. ABBB model was established by co-culture of primary cultured rat brain microvascular endothelial cells and astrocytes. The aBBB injury model was established by the induction of glucose and oxygen deprivation, the trans-endothelial electrical impedance, γ-glutamyl transpeptidase, alkaline phosphatase And lactate dehydrogenase to identify the damage of aBBB. The contents of ligustrazine, puerarin and nimodipine were determined by high performance liquid chromatography (HPLC), and the aBBB drug permeation concentration was calculated under OGD. Results: Compared with the control group, the TEER value in the model group decreased (P <0.01), the ligustrazine group, ligustrazine and puerarin combined group, nimodipine group-γGT and AKP increased (P <0.01) Compared with puerarin group, Ligustrazine and puerarin significantly increased AKP alone (P <0.01), and there was significant difference between Nimodipine and PGE (P <0.05) Increasing TEER, increasing -γGT, AKP content and decreasing LDH had synergistic effects. Compared with Ligustrazine and Puerarin, the permeation concentrations of Ligustrazine and puerarin in the culture medium of aBBB induced by OGD were both increased. CONCLUSION: Tetramethylpyrazine and puerarin are better than aBBB in combination with puerarin, which may be a protective effect on aBBB, which may be related to reducing the permeability of aBBB induced by OGD.