脑脉Ⅱ号口服液减轻高血压脑出血大鼠神经细胞凋亡的作用及机制研究

Reducing Effect of Naomai Ⅱ Oral Liquor on Apoptosis in Neuronal Cells Following Hypertensive Intracerebral Hemorrhage in Rats and Study on Its Mechanism

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 Naomai Ⅱ oral liquor on concentration of cytosolic free calcium （[Ca2＋ ]i） and apoptosis in the perihematomal brain tissue after hypertensive intracerebral, hemorrhage （HICH） in rats. Methods Eighty- five Sprague- Dawley rats were randomly assigned to sham - operation group, control group, and Chi- nese herbs compound group. HICH was induced in two- kidney, two clip renovascular hypertensive rat by injection of collage- nase into the right caudate nucleus. 8 ml of Naomai Ⅱ oral liquor or distilled water was intragastricly administered to rats 2 h after injection and re-administered until sacrificed. The rats＇ brains were removed at 6, 12, 24, 72, and 168 h after injec- tion. The apoptotic percentage in the perihematomal brain was determined by flow cytometry, and the [Ca2＋ ]i in synapto- somes was determined by Fura- 2/AM fluoremetry technique. Results The apoptotic percentage in the rats of control group increased significantly at 6 h （P〈0.01）, peaked at 72 h, decreased at 168 h after injection compared with sham- op- eration rats, the changes of [Ca2＋ ]i paralled that of apoptotic percentage. The apoptotic percentage and [Ca2＋ ]i in the rats of Chinese herbs compound group were significantly lowered than those in the rats of control group （P〈 0.05）. Conclusions Naomai Ⅱ oral liquor treatment can reduce neuronal apoptosis after HICH in rats; and its pharmacological mechanism may be related to suppressing the Ca2 ＋ overload in neurons.