中药提取物DM-1对阿尔茨海默病小鼠能量代谢及中枢胆碱能神经功能的影响

Effect of Chinese herb extract DM-1 on energy metabolism and central cholinergic nerve function in Alzheimer’s model mice

目的观察中药提取物DM-1对D-半乳糖联合三氯化铝(AlCl3)致拟阿尔茨海默病(AD)小鼠能量代谢及中枢胆碱能神经功能的影响。方法将70只昆明小鼠随机分为空白组、模型组、阳性对照组和低、高剂量对照组及低、高剂量实验组,每组10只。通过D-半乳糖皮下注射联合AlCl3灌胃共12周,制备小鼠AD模型。从第13周开始,空白组和模型组均按10 mL·kg^-1剂量灌胃0.9%NaCl;阳性对照组按10 mL·kg^-1剂量灌胃给予0.1 mg·mL^-1多奈哌齐;低、高剂量对照组均按10 mL·kg^-1剂量分别灌胃给予101和406 mg·mL^-1 DM-1,同时皮下注射150 mg·kg^-1D-半乳糖溶液;低、高剂量实验组均按10 mL·kg^-1剂量分别灌胃给予101和406 mg·mL^-1 DM-1。7组小鼠每天给药1次,连续给药4周。用Morris水迷宫观察小鼠学习记忆功能,用酶联免疫吸附法检测小鼠海马组织丙酮酸脱氢酶(PDH),α-酮戊二酸脱氢酶(α-KGDHC)和细胞色素氧化酶(CCO)含量。结果给药4周后,空白组、模型组、阳性对照组和低、高剂量对照组及低、高剂量实验组的逃逸平台进入次数分别为(3.20±1.62),(2.00±0.87),(2.73±1.29),(1.90±0.57),(1.70±0.67),(2.10±0.88)和(2.50±1.58)次,有效区域运动时间分别为(3.90±1.70),(2.42±0.42),(2.73±0.62),(2.09±0.63),(1.99±0.33),(2.46±0.58)和(3.02±0.56)s,海马组织中PDH含量分别为(376.85±77.16),(258.99±84.51),(386.73±37.38),(298.99±31.49),(304.29±63.77),(339.26±108.25)和(371.44±52.73)ng·mL^-1,α-KGDHC含量分别为(99.45±19.04),(85.11±23.93),(115.05±18.57),(91.31±13.69),(93.07±15.21),(97.55±85.05)和(98.78±15.39)pg·mL^-1,CCO含量分别为(184.05±104.47),(121.53±38.49),(208.46±125.85),(135.68±66.97),(142.85±38.60),(173.86±84.99)和(175.06±57.54)ng·mL^-1。阳性对照组及低、高剂量实验组中的上述指标与模型组比较,差异均有统计学意义(P<0.05或P<0.01)。结论DM-1对AD小鼠模型学习记忆功能具有一定的改善作用,其机制可能与增加脑内糖代谢的水平,提高脑能量的供应,从而增强模型小鼠中枢胆碱能神经功能,促进乙酰胆碱的合成有关。

Objective To investigate the effects of Chinese herb extract DM-1 on dementia of Alzheimer’s disease(AD)type induced by D-galactose and aluminum trichloride(AlCl3).Methods Seventy Kunming mice(KM)were randomly divided into blank,model,positive control,control-L,control-H,experimental-L and experimental-H groups with 10 mouse per group.D-galactose subcutaneous injection combined with AlC l3 intragastrically to prepare mouse AD model for 12 weeks.From the 13 th week,blank and model groups were intragastrically administered 0.9%NaCl at a dose of 10 mL·kg^-1.The positive control group was intragastrically administered with a concentration of 0.1 mg·mL^-1 at a dose of 10 mL·kg^-1.Control-L and control-H groups were intragastrically administered with a dose of 10 mL·kg^-1 at the concentration of101 and 406 mg·mL^-1 of DM-1,respectively,and subcutaneous injection with 150 mg·kg^-1 of D-galactose solution.Experimental-L and experimental-H groups were intragastrically administered with a dose of 10 mL·kg^-1 at the concentration of 101 and 406 mg·mL^-1 of DM-1,respectively.Seven groups were administered once a day for4 weeks.Morris water maze was used to observe the learning and memory function of mice.The enzyme-linked immunosorbent assay was used to detect the contents of pyruvate dehydrogenase(PDH),α-ketoglutarate dehydrogenase(α-KGDHC),and cytochrome C oxidase(CCO).Results After administration 4 weeks,the number of escape platform entry in the blank,model,positive control,control-L,control-H,experimental-L and experimental-H groups was(3.20±1.62),(2.00±0.87),(2.73±1.29),(1.90±0.57),(1.70±0.67),(2.10±0.88)and(2.50±1.58),the effective area exercise time was(3.90±1.70),(2.42±0.42),(2.73±0.62),(2.09±0.63),(1.99±0.33),(2.46±0.58)and(3.02±0.56)s,the contents of PDH in the hippocampal tissue were(376.85±77.16),(258.99±84.51),(386.73±37.38),(298.99±31.49),(304.29±63.77),(339.26±108.25)and(371.44±52.73)ng·mL^-1,the contents ofα-KGDHC were(99.45±19.04),(85.11±23.93),(115.05±18.57),(91.31±13.69),(93.07±15.21),(97.55±85.05)and(98.78±15.39)pg·mL^-1,the contents of CCO were(184.05±104.47),(121.53±38.49),(208.46±125.85),(135.68±66.97),(142.85±38.60),(173.86±84.99)and(175.06±57.54)ng·mL^-1.The above-mentioned indexes in the positive control,experimental-L and experimental-H groups were significantly different from those in the model group(P<0.05 or P<0.01).Conclusion Chinese herb extract DM-1 has a certain improvement on the learning memory function of AD mice induced by D-galactose and AlC l3,its mechanism may be related to the increase of brain sugar metabolism and improvement of brain energy supply,there by enhancing the model mouse central cholinergic nerve function and promoting the synthesis of acetylcholine.