芍药苷对APP/PS1小鼠的神经细胞保护作用及机制研究

Protective effect of paeoniflorin on nerve cells in APP/PS1 mice and its mechanism

目的:探讨芍药苷(paeoniflorin,PF)通过抑制细胞凋亡通路而产生神经细胞保护作用的机制。方法:分别选用15只5月龄雄性APP/PS1非显性小鼠作为正常对照组,15只5月龄雄性APP/PS1双转基因小鼠为模型组和15只5月龄雄性APP/PS1双转基因小鼠为给药组(5 mg/kg的PF腹腔注射)。采用水迷宫实验检测各组小鼠的学习和记忆能力。采用TUNEL荧光染色法检测脑内神经细胞凋亡情况。采用Western Blot检测脑内皮层及海马区PI3K、Akt、p-PI3K、p-Akt、caspase-3、caspase-9、Bcl-2和Bax的蛋白表达水平,并用免疫组化分析caspase-3和caspase-9的蛋白表达水平及分布情况。结果:(1)与正常对照组相比,APP/PS1模型组小鼠的学习和记忆能力明显下降;与APP/PS1模型组相比,PF明显改善小鼠的学习和记忆能力。(2)与正常对照组相比,APP/PS1模型组小鼠脑内神经细胞凋亡明显增多,分布区域较广,而PF给药组小鼠凋亡细胞明显减少。(3)与APP/PS1模型组相比,PF给药组能显著下调促凋亡因子caspase-3、caspase-9和Bax的表达水平(P<0.05),同时上调抑凋亡因子pPI3K、p-Akt和Bcl-2的表达水平(P<0.05)。结论:PF可能通过激活PI3K/Akt通路而上调Bcl-2,下调caspase-9、caspase-3和Bax的蛋白表达水平,从而抑制神经细胞凋亡和保护神经细胞,以治疗神经退行性疾病。

AIM： To investigate the therapeutic and preventive effects of paeoniflorin（ PF） on APP/PS1 mice,and to explore the possible mechanism. METHODS： Fifteen male 5-month-old APP/PS1 non-dominant mice were chosen as normal control group,15 male 5-month-old APP/PS1 double transgenic mice were used as model group,and 15 male 5-month-old APP/PS1 double transgenic mice treated with 5 mg/kg PF by intraperitoneal injection were allocated in administation group. The learning and memory ability of the mice in each group was detected by Morris water maze. The apoptosis was assessed by TUNEL fluorescence staining. The protein expression of PI3K,Akt,p-PI3K,p-Akt,caspase-3,caspase-9,Bcl-2 and Bax in cerebral cortex and hippocampus was detected by Western Blot. The protein expression levels and distribution of caspase-3 and caspase-9 were detected by immunohistochemistry. RESULTS：（ 1） Compared with normal control group,the learning and memory ability declined in APP/PS1 model group. Compared with APP/PS1 model group,PF obviously improve the ability of learning and memory in mice.（ 2） Compared with normal control group,the apoptosis of nerve cells in APP/PS1 model group significantly increased and distributed in wider areas,while that in PF group was reduced（ P〈0. 05）.（ 3） Compared with APP/PS1 model group,PF could significantly lower pro-apoptotic factors,caspase-3,caspase-9 and Bax（ P〈0. 05）,and increase the expression of anti-apoptotic factors,p-PI3K,p-Akt and Bcl-2（ P〈0. 05）. CONCLUSION： PF can up-regulate the expression of Bcl-2 and down-regulate the expression levels of caspase-9,caspase-3 and Bax via the activation of PI3K/Akt pathway,thereby inhibiting the nerve cell apoptosis and protecting the nerve cells,so as to treat neurodegenerative diseases.