丹参注射液抗乳酸神经毒性作用及其机制研究

The protective effects of radix salviae miltiorrhizae on injured neurons by lactate acid and its mechanism

目的 研究丹参注射液抗乳酸神经毒性作用及其机制。方法 利用SD大鼠大脑皮质培养神经元 ,通过MTT法或台盼蓝法测定细胞活力 ;以Fura 2 /Am为荧光指示剂 ,测定细胞内游离钙离子浓度 ( [Ca2 +]i)。结果 乳酸对神经元具有毒性作用 ,pH下调到相应水平对神经元无影响 ,终浓度为 2 0、2 0 0g·L-1的丹参注射液可以减轻乳酸引起的神经元损伤 ,具细胞保护作用 ;与对照组、pH下调组比较 ,乳酸可以增加神经细胞的静息 [Ca2 +]i水平 ;2 0、2 0 0 g·L-1浓度的丹参注射液可以抑制乳酸引起的静息 [Ca2 +]i升高 ,在高浓度的条件下 ( 2 0 0 g·L-1) ,对氯化钾、谷氨酸刺激诱发的[Ca2 +]i升高有抑制作用。结论 乳酸的神经毒性作用与pH下调无关 ,乳酸作为一种有机酸 ,可以直接促进 [Ca2 +]i升高 ;丹参对乳酸引起的损伤具有保护作用 。

AIM To explore the protective effects of radix salviae miltiorrhizae (RSM) on damaged neurons by lactate acid (LA) and its mechanism. METHODS The activity of cultured neurons was measured by MTT assay and trypan blue staining. Intracellular free Ca 2+ concentration ( i) in neurons was measured by RF- 5 000 double-wave-length fluorospectrophotometer, with Fura-2/Am as Ca 2+ indicator. RESULTS RSM could obviously attenuate the injury induced by LA on cortical neurons at certain concentration (20?200 g·L -1). The resting i of neurons raised with the add of LA, but it might be not related with the dropping of pH value. We also found RSM inhibited i elevation induced by LA or potassium chloride (KCl) and gutamate (Glu). CONCLUSIONS Taken together these results suggested that the damage of LA on neurons was not induced by the dropping of pH value, it can ease the inflow of Ca 2+ and contribute to the overload of Ca 2+. RSM has the protective effect on neurons damaged by LA. The mechanism of which may be correlated to its Ca 2+ antagonistic action.