金银花及其复方制剂对铝致阿尔茨海默症小鼠的干预作用研究

Study on the intervention of honeysuckle and its compound preparation on aluminum-induced Alzheimer’s disease in mice

目的探讨中药金银花及其复方制剂对铝致阿尔茨海默症(AD)小鼠的干预作用。方法将健康小白鼠随机分成正常组、模型组、金银花组和复方抗痴呆组,每组15只。模型组、金银花组和复方抗痴呆组采用D-半乳糖+氯化铝(AlCl 3)连续腹腔注射90 d建立AD小鼠模型,自造模第71天开始,金银花组和复方抗痴呆组分别给予金银花原液、复方抗痴呆口服液原液各0.15 mL加蒸馏水稀释至0.3 mL灌胃,正常组和模型组给予等量蒸馏水灌胃,均1次/d,连续20 d。采用Y型迷宫刺激器检测各组小白鼠的学习记忆能力,取血清检测生化指标[三酰甘油(TG)、总胆固醇(TC)、尿素氮(BUN)、谷丙转氨酶(ALT)、总蛋白(TP)],取脑组织检测乙酰胆碱酯酶(AchE)活力、超氧阴离子自由基清除率、铝(Al^(3+))及丙二醛(MDA)含量,HE染色观察脑组织病理变化。结果模型组小白鼠Y型迷宫刺激检测测试次数和记忆错误次数均明显多于正常组(P均<0.05);金银花组小白鼠记忆错误次数和复方抗痴呆组测试次数、记忆错误次数均明显少于模型组(P均<0.05),且复方抗痴呆组测试次数、记忆错误次数均明显少于金银花组(P均<0.05)。4组间血清TG、TC、BUN、TP水平比较差异均无统计学意义(P均>0.05);复方抗痴呆组血清ALT水平明显低于模型组和金银花组(P均<0.05),金银花组与模型组比较差异无统计学意义(P>0.05)。模型组脑组织中AchE活力、超氧阴离子自由基清除率均明显低于正常组(P均<0.05),Al^(3+)含量明显高于正常组(P<0.05);金银花组和复方抗痴呆组脑组织中AchE活力、超氧阴离子自由基清除率均明显高于模型组(P均<0.05),Al^(3+)含量明显低于模型组(P<0.05);复方抗痴呆组脑组织中AchE活力明显高于金银花组(P<0.05),超氧阴离子自由基清除率和Al^(3+)含量明显低于金银花组(P均<0.05)。模型组大脑皮质少数神经细胞呈急性溃变、慢性进行性坏死改变,金银花组病变程度比模型组轻,复方抗痴呆组病变程度比金银花组轻。结论中药金银花及其复方制剂可通过提高脑AchE活力和抗氧化能力、促进排铝而改善AD小白鼠的学习记忆能力,且金银花复方制剂较其单方制剂效果明显,可能与多种有效成分协同发挥作用有关。

Objective It is to explore the intervention effect of honeysuckle and its compound preparation on aluminum induced Alzheimer's disease(AD)in mice.Methods The healthy mice were randomly divided into normal group,model group,honeysuckle group and compound anti-dementia group,15 mice in each group.The model group,honeysuckle group and compound anti-dementia group were given D-galactose+aluminum chloride(AlCl 3)by continuous intraperitoneal injection for 90 days to establish AD mouse models.Since the 71st day of modeling,the honeysuckle group and compound anti-dementia group were respectively given the original liquid of honeysuckle and the original liquid of compound anti-dementia oral liquid 0.15 mL each diluted with distilled water to 0.3 mL by gavage,the normal group and the model group were given the same amount of distilled water by gavage,all once a day for 20 consecutive days.The learning and memory ability of the mice in each group was assessed by Y-type maze stimulator,the serum was taken to detect biochemical indicators[triacylglycerol(TG),total cholesterol(TC),urea nitrogen(BUN),alanine aminotransferase(ALT),total protein(TP)],brain tissue was taken to detect acetylcholinesterase(AchE)activity,superoxide anion free radical clearance rate,aluminum(Al^(3+))and malondialdehyde(MDA)content,and the pathological changes of brain tissue was observed by HE staining.Results The number of Y-maze stimulus detection tests and the number of memory errors in the model group were significantly higher than those in the normal group(all P<0.05);the number of memory errors in the honeysuckle group and the number of tests and memory errors in the compound anti-dementia group were significantly less than those in the model group(P<0.05),and the number of tests and memory errors in the compound anti-dementia group were significantly less than those in the honeysuckle group(P<0.05).There was no significant difference in serum TG,TC,BUN,TP between the 4 groups(all P>0.05);the serum ALT level of the compound anti-dementia group was significantly lower than that of the model group and the honeysuckle group(all P<0.05),and the difference between the model group and the honeysuckle group was was not statistically significant(P>0.05).The AchE activity and superoxide anion free radical clearance rate in the brain tissue of the model group were significantly lower than that of the normal group(all P<0.05),and the content of Al^(3+)was significantly higher than that of the normal group(P<0.05);the AchE activity and clearance rate in the brain tissue of the honeysuckle group and the compound anti-dementia group were significantly higher than those of the model group(P<0.05),and the content of Al^(3+)was significantly lower than that of the model group(P<0.05).The AchE activity in the brain tissue of the compound anti-dementia group was significantly higher than that of the honeysuckle group(P<0.05),and the clearance rate and Al^(3+)content were significantly lower than that of the honeysuckle group(all P<0.05).A few nerve cells in the cerebral cortex of the model group showed acute ulceration and chronic progressive necrosis.The honeysuckle group had less lesions than the model group,and the compound anti-dementia group had less lesions than the honeysuckle group.Conclusion Chinese medicine honeysuckle and its compound preparations can improve the learning and memory ability of AD mice by improving brain AchE activity,antioxidant capacity,and excretion of aluminum,and honeysuckle compound preparations are more effective than its single preparations,which may be related to the synergistic effect of multiple active ingredients.