牛黄醒脑丸对阿尔茨海默病模型小鼠海马区细胞凋亡的影响

Effect of Niuhuang-Xingnao pill on apoptosis in hippocampus of Alzheimer＆#39;s disease model mice

目的 探讨牛黄醒脑丸对阿尔茨海默病模型小鼠Bcl-2、caspase-3及C型-半胱氨酸天冬氨酸特异性蛋白酶-3（type C cysteine aspartate specific protease-3,C-caspase-3）表达的影响.方法 将50只APP/V717小鼠按随机区组分组法分为牛黄醒脑丸高、中、低剂量组,模型组,阳性对照组;另选取10只C57BL/6小鼠为空白对照组.牛黄醒脑丸高、中、低剂量组小鼠分别灌胃牛黄醒脑丸水溶液142、71、35.5 mg/kg;阳性对照组灌胃多奈哌齐溶液10 mg/kg;空白对照组及模型组灌胃等体积生理盐水.1次/d,连续给药60 d.采用Morris水迷宫实验检测各组小鼠的学习记忆能力.采用ELISA法检测小鼠海马组织SOD、GSH-Px及CAT含量;采用免疫印迹法检测小鼠海马组织Bcl-2、caspase-3、C-caspase-3表达.利用TUNEL染色法检测各组小鼠海马区细胞凋亡情况.结果 与模型组比较,牛黄醒脑丸高、中、低剂量组小鼠第Ⅲ象限停留时间[（36.58±4.57）s、（32.46±4.25）s、（29.71±4.26）s比（25.48±3.91）s]延长,穿越平台次数[（5.82±0.87）次、（4.59±0.76）次、（3.96±0.75）次比（3.27±0.53）次]增加（P〈0.05）;牛黄醒脑丸高、中、低剂量组小鼠海马GSH-Px[（161.14±14.01）U/mg、（150.76±13.16）U/mg、（143.17±12.54）U/mg比（120.78±10.92）U/mg]、SOD[（14.25±1.82）U/mg、（11.17±1.65）U/mg、（7.24±1.02）U/mg比（3.12±0.31）U/mg]、CAT[（17.95±2.16）U/mg、（16.72±1.83）U/mg、（15.54±1.47）U/mg比（13.25±2.60）U/mg]水平升高（P〈0.05）;caspase-3[（1.13±0.13）、（1.25±0.15）、（1.41±0.17）比（1.49±0.20）]、C-caspase-3[（1.17±0.14）、（1.27±0.16）、（1.42±0.18）比（1.52±0.23）]表达降低,Bcl-2[（0.88±0.08）、（0.79±0.06）、（0.67±0.04）比（0.59±0.04）]表达升高（P〈0.05）.TUNEL染色结果表明,牛黄醒脑丸高、中、低剂量组仅观察到少量凋亡神经元.结论 牛黄醒脑丸可调节阿尔茨海默病模型小鼠海马组织Bcl-2、caspase-3及C-caspase-3表达,抑制海马区神经细胞凋亡.

Objective To study the the treatment effects of Niuhuang-Xingnao pill on Alzheimer＇s diseases mice and its influence on the levels of Bcl-2, caspase-3 and C-caspase-3. Methods 50 APP/V717 mice were divided into the positive control group, the model group,Niuhuang-Xingnao pill high, medium and low dosage groups. 10 C57BL/6 mice were selected as blank control. The high, medium and low dosage groups were given 142, 71 and 35.5 mg/kgNiuhuang-Xingnao pill, the positive control group was given 10 mg/kg donepezil, the blank group and model group were given same volume normal salin. 1 time/day, and continuous administration for 60 d. Morris water maze experiment was performed to test the learning and memory ability in mice. The levels of CAT, SOD and GSH-Px were detected by the method of ELISA, while Bcl-2, caspase-3 and C-caspase-3 in the groups were detected by western-blotting. The apoptosis of hippocampus of mice were observed by the method of TUNEL. Results Compared with the model group, the third square time （36.58 ± 4.57 s, 32.46 ± 4.25 s, 29.71 ± 4.26 s vs.25.48 ± 3.91 s） of high, middle and low dose groups were significantly longer, cross time of the table （5.82 ± 0.87, 4.59 ± 0.76, 3.96 ± 0.75 vs.3.27 ± 0.53） were significantly higher （P〈0.05）. In the high, middle and low dose groups, the levels of GSH-Px （161.14 ± 14.01 U/mg, 150.76 ± 13.16 U/mg, 143.17 ± 12.54 U/mg vs. 120.78 ± 10.92 U/mg）, SOD （14.25 ± 1.82 U/mg, 11.17 ± 1.65 U/mg, 7.24 ± 1.02 U/mg vs. 3.12 ± 0.31 U/mg）, CAT （17.95 ± 2.16 U/mg, 16.72 ± 1.83 U/mg, 15.54 ± 1.47 U/mg vs. 13.25 ± 2.60 U/mg） were significantly higher （P〈0.05）;caspase-3 （1.13 ± 0.13, 1.25 ± 0.15, 1.41 ± 0.17 vs. 1.49 ± 0.20）, C-caspase-3 （1.17 ± 0.14, 1.27 ± 0.16, 1.42 ± 0.18 vs.1.52 ± 0.23） significantly lower（P〈0.05）, Bcl-2 （0.88 ± 0.08, 0.79 ± 0.06, 0.67 ± 0.04vs. 0.59 ± 0.04） significantly higher（P〈0.05）.ConclusionsNiuhuang-Xingnao pill treatment of Alzheimer＇s disease in mice can effectively promote the expression of Bcl-2, caspase-3 and C-caspase-3 in the hippocampus of mice, inhibit the apoptosis of the cells in hippocampus.