摘要
目的:探讨葛花总黄酮对2型糖尿病大鼠认知功能障碍及海马中脑源性神经营养因子(BDNF)表达的影响。方法:将Wistar大鼠100只随机分为对照组、模型组、吡拉西坦组(10 mg/kg)、葛花总黄酮低剂量组(20 mg/kg)及葛花总黄酮高剂量组(40 mg/kg),每组20只。模型组、吡拉西坦组、葛花总黄酮低及高剂量组均建立2型糖尿病大鼠模型,并于造模成功后给予相应药物灌胃;对照组和模型组大鼠灌胃等体积0.9%氯化钠溶液;给药体积均为10 ml/kg,1日1次,持续给予140 d。给药结束后,通过Morris水迷宫试验评价大鼠认知功能;采用苏木精-伊红染色观察大鼠海马神经元病理结构变化;检测大鼠海马组织中BDNF mRNA和蛋白,白细胞介素4(IL-4)、白细胞介素8(IL-8)和肿瘤坏死因子α(TNF-α)蛋白水平。结果:(1)对照组大鼠海马区神经元细胞结构正常;模型组大鼠海马区神经元大量坏死,有明显炎症细胞浸润;吡拉西坦组、葛花总黄酮低及高剂量组大鼠海马神经元细胞坏死减少,且神经元细胞结构较为完整。(2)与对照组相比,模型组大鼠逃避潜伏期时间明显延长,IL-4、IL-8及TNF-α蛋白表达水平明显升高,经过原平台位置的次数明显减少,原平台象限停留时间明显缩短,BDNF mRNA和蛋白表达水平明显降低,上述差异均有统计学意义(P <0.05)。(3)与模型组相比,吡拉西坦组、葛花总黄酮低及高剂量组大鼠逃避潜伏期时间明显缩短,IL-4、IL-8及TNF-α蛋白表达水平明显降低,经过原平台位置的次数明显增多,原平台象限停留时间明显延长,BDNF mRNA和蛋白表达水平升高,差异均有统计学意义(P <0.05);与葛花总黄酮低剂量组相比,葛花总黄酮高剂量组大鼠逃避潜伏期时间明显缩短,IL-4、IL-8及TNF-α蛋白表达水平明显降低,经过原平台位置的次数明显增多,原平台象限停留时间明显延长,BDNF mRNA和蛋白表达水平明显升高,差异均有统计学意义(P <0.05)。(4)与吡拉西坦组相比,葛花总黄酮低剂量组大鼠逃避潜伏期时间明显延长,IL-4、IL-8及TNF-α蛋白表达水平明显升高,经过原平台位置的次数明显减少,原平台象限停留时间明显缩短,BDNF mRNA和蛋白表达水平明显降低,差异均有统计学意义(P <0.05);葛花总黄酮高剂量组大鼠逃避潜伏期时间、原平台象限停留时间,IL-4、IL-8及TNF-α蛋白表达水平,经过原平台位置的次数,BDNF mRNA和蛋白表达水平与吡拉西坦组相比,差异均无统计学意义(P> 0.05)。结论:葛花总黄酮对2型糖尿病大鼠认知功能障碍的疗效明显,其机制与葛花总黄酮促进BDNF mRNA和蛋白表达,降低IL-4、IL-8及TNF-α表达水平,减轻胰岛素抵抗以及炎症反应有关。
OBJECTIVE:To investigate the effects of pueraria lobata total flavonoids on cognitive dysfunction and brain-derived neurotrophic factor(BDNF)expression in hippocampus of type 2 diabetes mellitus(T2DM)rats.METHODS:Totally 100 Wistar rats were randomly divided into the control group,model group,piracetam group(10 mg/kg),pueraria lobata total flavonoids low(20 mg/kg)and high(40 mg/kg)dose group,with 20 cases in each group.T2DM rats were established in model group,piracetam group and each dose group of pueraria lobata total flavonoids.After successful modeling,rats were given corresponding drugs by gavage.The control group and the model group were given the same volume of normal saline by gavage,the volume of administration was 10 ml/kg,once a day,for 140 d.At the end of the experiment,the cognitive function of rats was evaluated by Morris water maze test;hematoxylin eosin(HE)staining was used to observe the pathological changes of hippocampal neurons in rats;mRNA and protein levels of BDNF,interleukin-4(IL-4),interleukin-8(IL-8)and tumor necrosis factor-α(TNF-α)protein in hippocampus were detected.RESULTS:(1)The neuron cell structure in the hippocampus of the control group was normal;the neurons in the hippocampus of the model group were massively necrotic with obvious inflammatory cell infiltration;the necrosis of hippocampal neurons in the piracetam group and the low and high dose group of pueraria lobata total flavonoids decreased,and the neuronal cell structure was relatively complete.(2)The escape latency time in the model group were longer than that in the control group,the expression levels of IL-4,IL-8 and TNF-αprotein in the model group were higher than those in the control group,while the times of passing through the original platform position,staying time in the original platform quadrant,BDNF mRNA and protein expression levels were lower than those in the control group,with statistically significant differences(P<0.05).(3)The escape latency time in the piracetam group and pueraria lobata total flavonoids low and high dose group were shorter than that in the model group,the expression levels of IL-4,IL-8 and TNF-αprotein in the piracetam group and pueraria lobata total flavonoids low and high dose group were lower than those in the model group,while the times of passing through the original platform position,staying time in the original platform quadrant,BDNF mRNA and protein expression levels were higher than those in the model group,with statistically significant differences(P<0.05).The escape latency time in the pueraria lobata total flavonoids high dose group was shorter than that in the pueraria lobata total flavonoids low dose group,the expression levels of IL-4,IL-8 and TNF-αprotein in the pueraria lobata total flavonoids high dose group were lower than those in the pueraria lobata total flavonoids low dose group,while the times of passing through the original platform,staying time in the original platform quadrant,BDNF mRNA and protein expression levels were higher than those in the pueraria lobata total flavonoids low dose group,with statistically significant differences(P<0.05).(4)The escape latency time in the pueraria lobata total flavonoids low dose group was longer than that in the piracetam group,the expression levels of IL-4,IL-8 and TNF-αprotein in the pueraria lobata total flavonoids low dose group were higher than those in the piracetam group,while the times of passing through the original platform,staying time in the original platform quadrant,BDNF mRNA and protein expression levels were lower than those in the piracetam group,with statistically significant differences(P<0.05).The escape latency time,IL-4,IL-8,TNF-αprotein expression levels,times of passing through the original platform,staying time in the original platform quadrant,BDNF mRNA and protein expression levels in the pueraria lobata total flavonoids high dose group were similar to those in the piracetam group(P>0.05).CONCLUSIONS:The protective effects of pueraria lobata total flavonoids on cognitive dysfunction in T2DM rats is related to the mechanism that the total pueraria lobata total flavonoids can promote the mRNA and protein levels of BDNF,reduce the expression levels of IL-4,IL-8 and TNF-α,and alleviate the insulin resistance and inflammation.
作者
徐琳
李建霞
赵维娟
许雪廷
李娜
周燕
洪福娣
XU Lin;LI Jianxia;ZHAO Weijuan;XU Xueting;LI Na;ZHOU Yan;HONG Fudi(Dept.of Pharmacy,Beijing Chaoyang Emergency Medical Center,Beijing 100000,China;Dept.of Pharmacology,Seventh Medical Center of Chinese PLA General Hospital,Beijing 100700,China)
出处
《中国医院用药评价与分析》
2020年第11期1333-1336,1339,共5页
Evaluation and Analysis of Drug-use in Hospitals of China
基金
北京市科学技术委员会科研计划项目(No.Z171200002115003)。
关键词
葛花总黄酮
2型糖尿病
大鼠
认知功能障碍
脑源性神经营养因子
Pueraria lobata total flavonoids
Type 2 diabetes mellitus
Rats
Cognitive dysfunction
Brain-derived neurotrophic factor
