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目的探讨中药复方制剂脑脉Ⅱ号口服液对高血压脑出血大鼠血肿周围组织神经细胞内游离钙离子浓度([Ca2+]i)和细胞凋亡的影响。方法将85只大鼠随机分为假手术组、对照组和中药组。向双肾双夹肾血管性高血压大鼠右侧尾状核注入胶原酶诱导脑出血。注射后2 h开始,每天灌胃给予脑脉Ⅱ号口服液(8 ml/d)或等容积蒸馏水直至处死。注射后6、12、24、72、168 h取脑,用流式细胞术检测细胞凋亡百分率,用Fura-2/AM荧光标记法测定突触体内[Ca2+]i。结果与假手术组比较,对照组细胞凋亡百分率在注射6 h后显著增加(P<0.01),72 h达到高峰,168 h后明显下降;突触体内[Ca2+]i的变化规律与细胞凋亡百分率的变化规律类似。脑脉Ⅱ号口服液治疗后,中药组细胞凋亡百分率和突触体内[Ca2+]i均显著低于对照组(P<0.05)。结论脑脉Ⅱ号口服液能够降低高血压脑出血大鼠神经细胞凋亡发生率,其机制可能与抑制神经细胞内钙超载有关。
Objective To explore the effect of compound herb preparation of Naomai II oral liquid on intracellular free calcium concentration ([Ca2+]i) and apoptosis of hematoma in hypertensive intracerebral hemorrhage rats. Methods Eighty-five rats were randomly divided into sham operation group, control group and Chinese medicine group. Collagenase was injected into the right caudate nucleus of double-kidney double-clip renovascular hypertensive rats to induce cerebral hemorrhage. Two hours after the injection, the rats were given intragastric administration of Naomai II Oral Solution (8 ml/d) or equal volume of distilled water every day until they were sacrificed. The brains were harvested at 6, 12, 24, 72, and 168 h after injection. The percentage of apoptosis was measured by flow cytometry, and [Ca 2+] i in synaptosomes was measured by Fura-2/AM fluorescence labeling. Results Compared with the sham-operated group, the percentage of apoptosis in the control group increased significantly after 6 h injection (P<0.01), peaked at 72 h, and decreased significantly after 168 h; the change of [Ca2+]i and cell decline in synaptosomes. The change in percent mortality is similar. After treatment with Naomai II oral liquid, the percentage of apoptosis and [Ca2+]i in synaptophysin were significantly lower in the Chinese herb group than in the control group (P<0.05). Conclusion Naomai II oral liquid can reduce the incidence of neuronal apoptosis in rats with hypertensive intracerebral hemorrhage. The mechanism may be related to the inhibition of calcium overload in nerve cells.