醒脑启智胶囊对血管性痴呆小鼠海马组织碱性成纤维细胞生长因子mRNA表达的影响

Effect of xingnao qizhi capsule on the expression of basic fibroblast growth factor mRNA of hippocampal tissue in mice with vascular dementia

目的:观察醒脑启智胶囊对血管性痴呆小鼠学习和记忆功能及海马组织碱性成纤维细胞生长因子mRNA表达、脑组织病理学改变的影响。方法:实验于2004-12/2005-10在河北医科大学西校区完成。①取健康雄性5周龄昆明小鼠120只。将小鼠随机分为6组:假手术组,模型组,醒脑启智胶囊液高剂量组,醒脑启智胶囊液低剂量组,银杏叶液组,尼莫地平组,每组20只。②采用双侧颈总动脉反复结扎脑缺血再灌注的方法制备血管性痴呆小鼠模型。术后第2天进行分组干预。醒脑启智胶囊液高、低剂量组:灌服醒脑启智胶囊液(河北医科大学生产,由石菖蒲、川芎、橘络、枸杞子组成,为水煎醇沉工艺制作胶囊,每克浸膏相当于中药生药6.24g,实验时用0.5%羧甲基纤维素钠配成184g/L和92g/L溶液)1.84,0.92g/kg;银杏叶液组:灌服银杏叶液(每片含银杏叶提取物50mg,广西半宙制药股份有限公司生产,批号为20040902,实验时用0.5%羧甲基纤维素钠配制成浓度为2.5g/L的混悬液)0.05g/kg;尼莫地平组:灌服尼莫地平液(石家庄市华龙药业股份有限公司生产,批号为20041017,20mg/片)0.04g/kg,假手术组,模型组:均灌服生理盐水10mL/kg,1次/d,治疗7d。③采用电子水迷宫测试学习和记忆成绩;采用苏木精-伊红染色法进行脑组织的病理形态学观察,并采用反转录聚合酶链反应检测实验小鼠海马组织碱性成纤维细胞生长因子mRNA的表达。④计量资料差异比较采用方差分析、LSD法检验。结果:造模过程中死亡49只,71只进入结果分析。①光镜下病理形态学显示:模型组小鼠脑组织海马区呈缺血性病理改变,各用药组小鼠病变轻于模型组。②小鼠学习成绩与记忆成绩:模型组明显差于假手术组(P<0.01);各用药组明显优于模型组(P<0.05～0.01)。各用药组间比较,差异不明显(P>0.05)。③小鼠脑组织海马区碱性成纤维细胞生长因子mRNA的相对表达:模型组明显高于假手术组(P<0.05);醒脑启智胶囊液高、低剂量组,针杏叶液组和尼莫地平组均明显高于模型组(P<0.01);醒脑启智胶囊液高剂量组明显高于醒脑启智胶囊液低剂量组、银杏叶液组和尼莫地平组(P<0.05～0.01),醒脑启智胶囊液低剂量组、银杏叶液组、尼莫地平组之间比较,差异不明显(P>0.05)。结论:醒脑启智胶囊可明显改善血管性痴呆小鼠学习和记忆功能,其作用机制之一是调节海马组织碱性成纤维细胞生长因子mRNA的表达,减轻缺血再灌注损伤。

AIM： To investigate the effects of xingnao qizhi capsule on learning and memory function, expression of basic fibroblast growth factor （bFGF） mRNA of hippocampal tissue and histopathological changes of brain .tissue in mice with vascular dementia （VD）. METHODS： The experiment was completed at the western area of Hebei Medical University from December 2004 to October 2005. ① Totally 120 male Kunming mice were randomly divided into 6 groups： sham-operation group, model group, high-dose xingnao qizhi group, low-dose xingnao qizhi group, ginkgo leaf group and nimodipine group with 20 in each group. ② VD mice models were established with cerebral ischemia repeatedly ligated on bilateral common carotid artery. Grouping intervention was performed on the second day after operation. High and low dosage xingnao qizhi group： Mice in this group were perfused with 1.84 and 0.92 g/kg xingnao qizhi capsule （produced in Hebei Medical University, consisting of shichangpu, ehuanxiong, juluo and gouqizi; 6.24 g raw drug per extract; 184 g/L and 92 g/L solution were mixed with 0.5% carboxymethylcellulose sodium during experiment）; ginkgo leaf group： Mice in this group were pcrfused with 0.05 g/kg ginkgo leaf （50 mg ginkgo leaf extract per pill; Guangxi Banzhou Pharmacological Limited Company; batch number： 20040902; 2.5 g/L suspension was mixed with 0.5% carboxymethylcellulose sodium during experiment）; nimodipine group： Mice in this group were perfused with 0.04 g/kg nimodipine （Shijiazhuang Hualong Pharmacological Limited Company; batch number： 20041017, 20 mg/pill）; sham-operation group and model group： Mice in both groups were perfused with 10 mL/kg saline once a day for 7 days. ③ Results of learning and memory were assayed with electric water maze; pathomorphological changes in brain tissue were observed with haematine-eosin staining; and expression of bFGF mRNA of hippocampal tissue in mice were detected with reverse transcription polymerase chain reaction. ④Measurement data were compared with analysis of variance and LSD method. RESULTS： Totally 49 mice died during modeling, and other 71 mice entered the final analysis. ① Pathomorphology under light microscope： Ischemic pathological changes were observed in hippocampus of brain tissue of mice in model group, and lesion in each drug group was lighter than that in model group.②Results of learning and memory： Results of mice in model were lower than those in sham operation group （P 〈 0.01）; but those in each drug group were superior to those in model group （P 〈 0.05-0.01）. There was not significant difference among drug groups （P 〉 0,05）, ③ Relative expression of bFGF mRNA in hippoeampal tissue： That in model group was higher than that in sham-operation group （P 〈 0,05）; that in high-dosage and low-dosage xingnao qizhi groups, ginkgo leaf group and nimodipine group was higher than that in model group （P 〈 0.01）; that in high-dosage xingnao qizhi group was higher than that in low-dosage xingnao qizhi group, ginkgo leaf group and nimodipine group （P 〈 0.05-0.01）; there were not significant differences among low-dosage xingrtao qizhi group, ginkgo leaf group and nimodipine group （P 〉 0.05）. CONCLUSION： Xingnao qizhi capsule can improve learning and memory function of VD mice. The mechanisms are regulating the expression of bFGF mRNA of hippocampals tissue and relieving ischemia-repeffusion injury.