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目的:探讨通络化痰胶囊对脑缺血损伤大鼠神经细胞凋亡的影响和机制。方法:98只SD大鼠通过永久性大脑中动脉闭塞(pMCAO)制备脑缺血损伤模型,待大鼠清醒后,根据Longa评分(≥2分)随机分为7组,分别为正常对照组、假手术组、模型组、尼莫地平组、通络化痰高、中、低剂量组。于术后第1天开始分别给予蒸馏水(正常对照组、假手术组、模型组)、尼莫地平(32.4 mg·kg-1)和通络化痰胶囊高、中、低剂量组(648,324,162 mg·kg-1)灌胃治疗,并于术后第1,2,3,5,7天行Narrow-Alley Corner Test检测脑缺血损伤大鼠神经功能的改变,术后第8天行4%多聚甲醛(pH 7.0)灌注固定后断头取脑,观察脑组织病理学的改变,利用TUNEL法检测缺血半暗带神经细胞凋亡水平的变化,利用免疫组化技术测定缺血半暗带Bcl-2相关x蛋白(Bax)和B细胞淋巴瘤/白血病-2(Bcl-2)表达水平的改变。结果:模型组大鼠Narrow-Alley Corner Test得分较正常对照组和假手术组显著增加(P<0.01),尼莫地平组和通络化痰胶囊组得分较模型组降低(P<0.01,P<0.05)。缺血后大鼠脑组织损伤明显,尼莫地平组和通络化痰胶囊组的损伤减轻。模型组较正常对照组和假手术组神经细胞凋亡增多、缺血半暗带Bax和Bcl-2表达增加(P<0.01)。尼莫地平组和通络化痰胶囊组较模型组神经细胞凋亡减轻、缺血半暗带Bax表达减少,Bcl-2表达增强(P<0.01)。结论:通络化痰胶囊可促进脑缺血损伤大鼠损伤后神经功能障碍的恢复,减轻神经元病理损害,其机制可能与其促进缺血半暗带Bcl-2表达,抑制Bax表达,从而减少细胞凋亡水平有关。
Objective: To investigate the effect of Tongluo Huatan capsule on neuronal apoptosis in rats with cerebral ischemia injury and its mechanism. Methods: One hundred and forty-eight SD rats were subjected to permanent cerebral occlusion (pMCAO) to establish a model of cerebral ischemia. After awake, the rats were randomly divided into 7 groups according to Longa score (≥2 points): normal control group, Sham operation group, model group, nimodipine group, Tongluo phlegm high, medium and low dose groups. Rats in the control group, sham operation group, model group, nimodipine (32.4 mg · kg-1) and Tongluo Huatan capsule high, medium and low dose groups (648,324,162 mg · Kg-1) intragastrically. Narrow-Alley Corner Test was performed on the 1st, 2nd, 3rd, 5th and 7th day after operation to detect the neurological changes in rats with cerebral ischemia. On the 8th day after operation, 4% Paraformaldehyde (pH 7.0) was perfused and decapitated to observe the changes of brain pathology. The changes of neuronal apoptosis in ischemic penumbra were detected by TUNEL method. With Bcl-2-related protein (Bax) and B-cell lymphoma / leukemia-2 (Bcl-2) expression changes. Results: The scores of Narrow-Alley Corner Test in model group were significantly higher than those in normal control group and sham operation group (P <0.01). The scores of Nimodipine and Tongluo Huatan Capsule were lower than those in model group (P <0.01, P <0.05). The ischemic rat brain tissue injury significantly, nimodipine group and Tongluo Huatan capsule group reduce the damage. Compared with normal control group and sham operation group, neuronal apoptosis increased in model group, and expression of Bax and Bcl-2 increased in ischemic penumbra (P <0.01). Compared with model group, Nimodipine and Tongluo Huatan Capsule showed less neuronal apoptosis, decreased Bax expression and increased Bcl-2 expression in ischemic penumbra (P <0.01). Conclusion: Tongluo Huatan Capsule can promote the recovery of neurological dysfunction and reduce the pathological damage of neurons after traumatic brain injury in rats, which may be related to the promotion of Bcl-2 expression and the decrease of Bax expression in ischemic penumbra, thus reducing The level of apoptosis related.