中药复方脑益康干预阿尔茨海默病模型小鼠脑组织神经生长因子及脑源性神经营养因子mRNA的表达

Effect of Chinese herb compound naoyikang on the mRNA expression of nerve growth factor and brain-derived neurotrophic factor in mice model of Alzheimer disease

目的:观察中药复方脑益康对阿尔茨海默病模型小鼠脑组织脑神经生长因子、脑源性神经营养因子mRNA表达的影响,。方法:①实验于2004-10/2005-03在南通大学基础医学院病理生理学实验室完成。②采用D-半乳糖和亚硝酸钠腹腔注射建立阿尔茨海默病小鼠模型,ICR小鼠120只,采用随机数字法分为6组,正常对照组,模型对照组,脑复康对照组[0.4g/(kg·d)],脑益康小、中、大剂量组[(2.4,7.2,24g/(kg·d)]。脑益康由制首乌、巴戟天等组成(南通市中医院制剂室制成颗粒剂,每克颗粒剂含生药1.98g)。模型对照组、各给药组腹腔注射10g/LD-半乳糖120mg/(kg·d)和10g/L亚硝酸钠90mg/(kg·d),连续60d,正常对照组腹腔注射等容量的生理盐水。脑益康和脑复康均以5g/L羧甲基纤维素钠配制灌胃,连续60d。③每组随机取6只小鼠,取脑组织,采用反转录-聚合酶链反应方法检测脑组织中脑神经生长因子、脑源性神经营养因子mRNA的表达。结果:①脑益康小、中、大剂量组脑神经生长因子mRNA的表达与正常对照组比较差异无显著性(1.05±0.25,0.83±0.46,0.99±0.46,1.12±0.22,P>0.05);脑益康小、中、大剂量组脑源性神经营养因子mRNA的表达与正常对照组比较差异无显著性(0.73±0.20,0.49±0.12,0.63±0.15,0.32±0.22,P>0.05)。②模型对照组脑神经生长因子mRNA、脑源性神经营养因子mRNA显著高于正常对照组(1.98±0.60比1.12±0.22,1.32±0.37比0.32±0.22,P<0.05)。③与模型对照组脑神经生长因子mRNA和脑源性神经营养因子mRNA比较,阳性药脑复康对照组、脑益康小、中、大剂量组表达明显减少(P<0.05)。结论:脑益康提高阿尔茨海默病小鼠学习记忆作用的部分机制与其保护胆碱能神经元、改善神经生长因子、脑源性神经营养因子的逆行运输有关。

AIM： To investigate the effect of Chinese herb compound nooyikang on the mRNA expression of nerve growth factor （NGF） and brain derived neurotrophic factor （BDNF） in mice model of Alzheimer disease. METHODS： ①The experiment was conducted in the Laboratory of Pathophysiology, School of Basic Medical Sciences, Nantong University from October 2004 to March 2005. ②The mice models of Alzheimer disease were established by intraperitoneal injection of D-galactosamine and natrium nitrosum. 120 ICR mice were randomly divided into normal control group, model control group, naofukang control group （0.4 g/kg per day）, and low, middle and high dose naofukang groups （2.4, 7.2, 24 g/kg per day）. Naoyikang was composed of shouwu, bajitian and so on （produced by the Pharmaceutical Room of Nantong Hospital of Chinese Medicine,1.98 g crude drug in per g granule）. The model control group and all the treated groups were intraperitoneally injected with 10 g/L D-galactosamine 120 mg/kg per day and 10 g/L natrium nitrosum 90 mg/kg per day for 60 days; the norreal control group was injected with normal saline at the same dosage. Naoyikang and naofukang prepared with 5 g/L sodium carboxymethycellulose were intragastrically infused for 60 days. ③Six mice in each group were randomly selected to take the brain tissues. The mRNA expression of NGF and BDNF were tested by RT-PCR. RESULTS： ①Compared with the control group, the expression of NGF mRNA in low, middle and high doses naofukang groups had no significant difference [（1.05±0.25）, （0.83±0.46）, （0.99±0.46）, （1.12±0.22）, P 〉 0.05]; Compared with the control group, the expression of BDNF mRNA in low, middle and high doses naofukang groups had no significant difference [（0.73±0.20）, （0.49±0.12）, （0.63±0.15）, （0.32±0.22）, P 〉 0.05]. ②The expressions of NGF and BDNF mRNA of the model group were significantly higher than that of the normal control group [NGF mRNA （1.98±0.60）, （1.12±0.22）; BDNF mRNA （1.32±0.37）, （0.32±0.22）, P 〈 0.05].③Compared with the model group, the expressions of NGF and BDNF mRNA in the naofukang control group and low, middle and high doses naofukang groups were obviously decreased （P 〈 0.05）. CONCLUSION： Naoyikartg can improve part mechanism of learning and memory ability of mice with Alzheimer disease by protecting cholinergic neurons and improving the reversal transportation of NGF and BDNF.