当归有效组分防治阿尔茨海默病模型大鼠组方配伍优化的实验研究
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摘要
目的优化当归有效组分当归多糖、当归挥发油及阿魏酸防治阿尔茨海默病(AD)的最佳配伍。方法从150只SPF级Wistar大鼠中随机选取10只作为空白组、10只作为假手术组,其余大鼠均在脑立体定位下于海马CA1区左右两侧分别注射1μL Aβ_(25-35)复制AD大鼠模型,筛选造模成功的110只大鼠,随机分为模型组、当归组及当归组分配伍1～9组等11组,每组10只。造模第7天开始灌胃给药,空白组、假手术组与模型组均灌胃生理盐水,当归组灌胃当归水煎液,当归组分配伍各组灌胃对应药液,连续灌胃28 d。给药第24～27天进行Morris水迷宫定位航行试验,第28天进行空间探索试验。当归组分配伍组以当归多糖、当归挥发油、阿魏酸为考察因素,采用正交试验法,以各组大鼠逃避潜伏期和跨原平台次数为考察指标,优选当归有效组分防治AD的最佳配伍。结果与假手术组比较,模型组大鼠逃避潜伏期明显延长,跨原平台次数明显减少,差异均有统计学意义(P<0.05或P<0.01)。与模型组比较,各给药组大鼠逃避潜伏期均不同程度缩短,差异有统计学意义(P<0.05或P<0.01);当归组,当归组分配伍1,2,5,6,7,9组跨原平台次数明显增多,差异均有统计学意义(P<0.05或P<0.01)。与当归组比较,当归组分配伍1,4,5,6,7,8组逃避潜伏期明显缩短,差异均有统计学意义(P<0.05或P<0.01)。当归有效组分改善AD模型大鼠学习记忆能力的最佳配伍是1 m L组分配伍药液中含当归多糖0.017 6 g、当归挥发油0.004 4 m L、阿魏酸0.001 2 g。结论当归有效组分配伍能够有效地改善AD模型大鼠的学习记忆能力,对AD具有一定的防治作用。
Objective To optimize the best compatibility between Angelicae senensis polysaccharide and volatile oil as well as ferulic acid of the active components in Danggui( Radix Angelicae senensis) for the prevention and cure of Alzheimer's disease( AD).Methods Of 150 SPF Wistar rats,10 were randomly chosen as blank group,another 10 were chosen as sham operation group,and the rest of them were injected 1 μL Aβ_(25-35) into the left and right sides of the CA1 subfield of the hippocampal formation with stereotaxic apparatus to replicate AD rats model,and 110 successful rats model were screened and randomly divided into 11 groups,including model group,Danggui group and 1 to 9 Danggui component compatibility groups,with 10 rats in each. The rats in the blank group,shamoperation group and model group were given normal saline by intragastric administration on the 7 th day of modeling,the rats in the Danggui group were given Danggui water decoction by intragastric administration,and the rats in the Danggui component compatibility groups were given corresponding soup by intragastric administration for 28 consecutive days. The Morris water maze place navigation test was conducted on the 24 th to 27 th day of drug administration,and the spatial probe test was conducted on the 28 th day. In the Danggui component compatibility groups,the orthogonal experiment was adopted with polysaccharide,volatile oil and ferulic acid in Danggui as investigation factors,and with the rats' escape latency and the times of leaping over platform as investigation index to optimize the best compatibility of active components in Danggui for prevention and cure of AD. Results Compared with sham operation group,the escape latency significantly prolonged and the times of leaping over platform significantly decreased of rats in the model group,and the difference was statistically significant( P<0.05 or P<0.01). Compared with the model group,the escape latency significantly shortened in rats of each medication administration groups,and the difference was statistically significant( P<0.05 or P <0.01). In Danggui group and the 1~(st),2~(nd),5~(th),6~(th),7~(th) and 9~(th) Danggui component compatibility groups,the times of leaping over platform significantly increased,and the difference was statistically significant( P<0.05 or P<0.01). Compared with Danggui group,the escape latency significantly shortened in the 1~(st),4~(th),5~(th),6~(th),7~(th) and 8~(th) Danggui component compatibility groups,and the difference was statistically significant( P<0.05 or P<0.01).The best compatibility of active components of Danggui in improving learning and memory ability of AD rats model is 1 m L compatible soup containing Angelicae senensis polysaccharide 0.017 6 g and volatile oil 0.004 4 m L and ferulic acid 0.001 2 g. Conclusion The compatibility of active components of Radix Angelicae sinensis can effectively improve the learning and memory ability of AD rats model,and has certain prevention and cure effect on AD.
Objective To optimize the best compatibility between Angelicae senensis polysaccharide and volatile oil as well as ferulic acid of the active components in Danggui( Radix Angelicae senensis) for the prevention and cure of Alzheimer's disease( AD).Methods Of 150 SPF Wistar rats,10 were randomly chosen as blank group,another 10 were chosen as sham operation group,and the rest of them were injected 1 μL Aβ_(25-35) into the left and right sides of the CA1 subfield of the hippocampal formation with stereotaxic apparatus to replicate AD rats model,and 110 successful rats model were screened and randomly divided into 11 groups,including model group,Danggui group and 1 to 9 Danggui component compatibility groups,with 10 rats in each. The rats in the blank group,shamoperation group and model group were given normal saline by intragastric administration on the 7 th day of modeling,the rats in the Danggui group were given Danggui water decoction by intragastric administration,and the rats in the Danggui component compatibility groups were given corresponding soup by intragastric administration for 28 consecutive days. The Morris water maze place navigation test was conducted on the 24 th to 27 th day of drug administration,and the spatial probe test was conducted on the 28 th day. In the Danggui component compatibility groups,the orthogonal experiment was adopted with polysaccharide,volatile oil and ferulic acid in Danggui as investigation factors,and with the rats' escape latency and the times of leaping over platform as investigation index to optimize the best compatibility of active components in Danggui for prevention and cure of AD. Results Compared with sham operation group,the escape latency significantly prolonged and the times of leaping over platform significantly decreased of rats in the model group,and the difference was statistically significant( P<0.05 or P<0.01). Compared with the model group,the escape latency significantly shortened in rats of each medication administration groups,and the difference was statistically significant( P<0.05 or P <0.01). In Danggui group and the 1~(st),2~(nd),5~(th),6~(th),7~(th) and 9~(th) Danggui component compatibility groups,the times of leaping over platform significantly increased,and the difference was statistically significant( P<0.05 or P<0.01). Compared with Danggui group,the escape latency significantly shortened in the 1~(st),4~(th),5~(th),6~(th),7~(th) and 8~(th) Danggui component compatibility groups,and the difference was statistically significant( P<0.05 or P<0.01).The best compatibility of active components of Danggui in improving learning and memory ability of AD rats model is 1 m L compatible soup containing Angelicae senensis polysaccharide 0.017 6 g and volatile oil 0.004 4 m L and ferulic acid 0.001 2 g. Conclusion The compatibility of active components of Radix Angelicae sinensis can effectively improve the learning and memory ability of AD rats model,and has certain prevention and cure effect on AD.
引文
[1]吴红彦,王虎平.以肝脾论治老年性痴呆及逍遥散对老年痴呆模型小鼠学习记忆能力、中枢胆碱能神经元活性的影响[J].中国实验方剂学杂志,2010,16(5):168-170.
[2]吴红彦,李海龙,王虎平,等.大剂量当归对东莨菪碱致痴呆小鼠模型学习记忆及Ach E、Ch AT活性的影响[J].时珍国医国药,2013,24(3):552-554.
[3]杨植媛,吴红彦,李海龙,等.当归含药血清对阿尔茨海默病细胞模型损伤的保护作用[J].辽宁中医杂志,2014,41(1):164-167.
[4]PAXINOS G,WATSON C.The rat brain in stereotaxic coordinates[M].5th ed.San Diego:Elsevier Academic Press,2005.
[5]张杰,马云枝,沈晓明,等.复智胶囊对血管性痴呆模型大鼠学习记忆能力改善的作用机制[J].中国实验方剂学杂志,2014,20(2):128-132.
[6]戚晓利,徐秀芳,魏晓东.当归对D-半乳糖衰老模型小鼠抗氧化系统的研究[J].黑龙江医药科学,2003,26(1):223.
[7]杨静薇,廖维靖.当归对大鼠局灶性缺血脑组织细胞凋亡MAP2作用[J].辽宁中医杂志,2001,28(8):505-506.
[8]PARK S J,JUNG J M,LEE H E,et al.The memory ameliorating effects of INM-176,an ethanolic extract of Angelica gigas,against scopolamine-or Aβ1-42-induced cognitive dysfunction in mice[J].J Ethnopharmacol,2012,143(2):611-620.
[9]魏明刚,张玲,李凤玲,等.加味当归补血汤对阿霉素肾病大鼠肾脏足细胞保护机制的研究[J].中国中西医结合杂志,2012,32(8):1077-1082.
[10]吴丽蓉,罗勇.丁基苯酞对神经系统疾病的作用及机制[J].中国康复理论与实践,2006,12(12):936.
[11]徐露,董志.当归多糖抗衰老作用的实验研究[J].激光杂志,2008,29(4):89-90.
[12]孙文平,罗红,杨光,等.当归多糖激发免疫反应的特征研究[J].大连医科大学学报,2009,31(3):262-264.
[13]肖吉元,杨扶德,白银亮,等.当归挥发油对吗啡依赖小鼠戒断症状及单胺类神经递质的影响[J].中国医院药学杂志,2015,35(13):1188-1190.
[14]刘露丝,岳美颖,李文兵,等.当归油与川芎油的化学成分及其对急性化学性脑损伤保护作用差异研究[J].中药药理与临床,2016,32(6):105-108.
[15]黄浩,马增春,王宇光,等.阿魏酸对脂多糖损伤的PC12细胞和大鼠海马神经元细胞的保护作用[J].中国药理学与毒理学杂志,2016,30(4):330-337.
[16]王玥,王旭,于嵩,等.阿魏酸对阿尔茨海默病转基因小鼠脑内氧化应激和凋亡相关蛋白的影响[J].天然产物研究与开发,2017(29):762-766.
[17]国家药典委员会.中华人民共和国药典[M].北京:中国中医药科技出版社,2015:133.
[2]吴红彦,李海龙,王虎平,等.大剂量当归对东莨菪碱致痴呆小鼠模型学习记忆及Ach E、Ch AT活性的影响[J].时珍国医国药,2013,24(3):552-554.
[3]杨植媛,吴红彦,李海龙,等.当归含药血清对阿尔茨海默病细胞模型损伤的保护作用[J].辽宁中医杂志,2014,41(1):164-167.
[4]PAXINOS G,WATSON C.The rat brain in stereotaxic coordinates[M].5th ed.San Diego:Elsevier Academic Press,2005.
[5]张杰,马云枝,沈晓明,等.复智胶囊对血管性痴呆模型大鼠学习记忆能力改善的作用机制[J].中国实验方剂学杂志,2014,20(2):128-132.
[6]戚晓利,徐秀芳,魏晓东.当归对D-半乳糖衰老模型小鼠抗氧化系统的研究[J].黑龙江医药科学,2003,26(1):223.
[7]杨静薇,廖维靖.当归对大鼠局灶性缺血脑组织细胞凋亡MAP2作用[J].辽宁中医杂志,2001,28(8):505-506.
[8]PARK S J,JUNG J M,LEE H E,et al.The memory ameliorating effects of INM-176,an ethanolic extract of Angelica gigas,against scopolamine-or Aβ1-42-induced cognitive dysfunction in mice[J].J Ethnopharmacol,2012,143(2):611-620.
[9]魏明刚,张玲,李凤玲,等.加味当归补血汤对阿霉素肾病大鼠肾脏足细胞保护机制的研究[J].中国中西医结合杂志,2012,32(8):1077-1082.
[10]吴丽蓉,罗勇.丁基苯酞对神经系统疾病的作用及机制[J].中国康复理论与实践,2006,12(12):936.
[11]徐露,董志.当归多糖抗衰老作用的实验研究[J].激光杂志,2008,29(4):89-90.
[12]孙文平,罗红,杨光,等.当归多糖激发免疫反应的特征研究[J].大连医科大学学报,2009,31(3):262-264.
[13]肖吉元,杨扶德,白银亮,等.当归挥发油对吗啡依赖小鼠戒断症状及单胺类神经递质的影响[J].中国医院药学杂志,2015,35(13):1188-1190.
[14]刘露丝,岳美颖,李文兵,等.当归油与川芎油的化学成分及其对急性化学性脑损伤保护作用差异研究[J].中药药理与临床,2016,32(6):105-108.
[15]黄浩,马增春,王宇光,等.阿魏酸对脂多糖损伤的PC12细胞和大鼠海马神经元细胞的保护作用[J].中国药理学与毒理学杂志,2016,30(4):330-337.
[16]王玥,王旭,于嵩,等.阿魏酸对阿尔茨海默病转基因小鼠脑内氧化应激和凋亡相关蛋白的影响[J].天然产物研究与开发,2017(29):762-766.
[17]国家药典委员会.中华人民共和国药典[M].北京:中国中医药科技出版社,2015:133.