脑出血后血肿周围神经元Cyt-c与Caspase-3的表达及不同中医治法的干预

Effects of Different Traditional Chinese Medicines on Expressions of Cyt-c and Activated Caspase-3 After Intracerebral Hemorrhage in Rats

目的观察大鼠脑出血后血肿周围神经元Cyt-c与活化Caspase-3的表达情况及不同中医治法的干预作用。方法用Ⅶ胶原酶脑内立体定位注射诱导大鼠脑出血模型,免疫组化检测血肿周围神经元Cyt-c的释放情况与活化Caspase-3的表达,同时用直尺测量不同时间点血肿的最大直径。结果大鼠脑出血后血肿周围神经元于6h即有Cyt-c释放;1d后阳性细胞表达最多;3d后阳性细胞数减少;5d后阳性细胞继续减少,与假手术组比较差异明显(P<0.01);血肿周围于6h出现Caspase-3阳性表达细胞,1d阳性细胞计数达高峰,3d出现轻度下降,5d明显下降,与假手术组比较差异明显(P<0.01);采取不同中医治法治疗后Cyt-c与Caspase-3均有所下降,与其他各组比较,平肝熄风汤组治疗后下降最明显(P<0.01)。结论脑出血后血肿周围神经元Cyt-c与活化Caspase-3表达明显上调,采取熄风、泻火、祛痰、化瘀以及风火痰瘀同治等治法均能抑制Cyt-c与Caspase-3的表达,阻止神经元的凋亡,以平肝熄风汤组最为明显。

Objective To study the effects of different traditional Chinese medicines on the expressions of Cyt- c and activated Caspase- 3 in neuron around hematoma after intracerebral hemorrhage （ICH） in rat models, and the change of hematoma. Methods Ⅻ type collagenase was stereotaxically injected into the globas pallidus of rats to build the ICH model. Immunohistochemical method was used to examine the expressions of Cyt - c and activated Caspase - 3 after ICH in rats at different time points, and the haematoma was measured at its biggest diameter. Results Cyt - c began to release in the neurons around the hematoma after ICH in rats at 6 h after operation. One day thereafter, the number of positive cells reached to peak value; 3 days thereafter, the staining became weakening, the positive cells decreased; and 5 days thereafter, the positive cells continue to decrease, which were varied significantly as compared with the sham- operated group （P〈 0.01 ）. The activated Caspase- 3 in cytoplasm appeared around the hematoma 6 h after the operation, reached peak value I day thereafter, then declined slightly 3 days thereafter, and declined significantly 5 days thereafter as compared with the sham- operated group; the difference was significant （P〈 0.01 ）. The number of positive cells decreased after treatment with different traditional Chinese medicine, and the Pingganxifeng decoction group was superior to the other groups（P〈 0.01 ）. Conclusions The release of Cyt - c and the activated Caspase - 3 increase significantly in the neurons around the hematoma after ICH. The expressions of activated Cyt - c and Caspase - 3 can be depressed through suppressing wind, removing phlegm, inducing resuscitating, and promoting blood circulation by removing blood stasis, etc. The results suggest that the Pingganxifeng decoction can prohibit neuronal apoptosis, and its efficacy is the best.