Protective effect of tetrahydroxy stilbene glucoside on learning and memory by regulating synaptic plasticity

Damage to synaptic plasticity induced by neurotoxicity of amyloid-beta is regarded to be one of the pathological mechanisms of learning and memory disabilities in Alzheimer＇s disease patients. This study assumed that the damage of amyloid-beta to learning and memory abilities was strongly associated with the changes in the Fyn/N-methyl-D-aspartate receptor 2B （NR2B） expression. An APP695V7171 transgenic mouse model of Alzheimer＇s disease was used and treatment with tetrahydroxy-stilbene glucoside was administered intragas- trically. Results showed that intragastric administration of tetrahydroxy-stilbene glucoside improved the learning and memory abilities of the transgenic mice through increasing NR2B receptors and Fyn expression. It also reversed parameters for synaptic interface structure of gray type I. These findings indicate that tetrahydroxy stilbene glucoside has protective effects on the brain, and has prospects for its clinical application to improve the learning and memory abilities and treat Alzheimer＇s disease.

Tetrahydroxy Stilbene Glucoside Ameliorates Cognitive Impairments and Pathology in APP/PS1 Transgenic Mice

Cognitive impairment is the main clinical manifestation of Alzheimer's disease(AD),and amyloid-β(AB)deposition and senile plaques are the characteristic neuropathological hallmarks in AD brains.This study aimed to explore the effect and mechanism of tetrahydroxy stilbene glucoside(TSG)on cognitive function in APP/PS 1 mice during long-term administration.Here,we treated APP/PS1 model mice of AD with different doses of TSG(50 mg/kg and 100 mg/kg)for 5 to 17 months by gavage,and we further observed whether TSG could ameliorate the cognitive decline in APP/PS1 mice using behavioral tests,and investigated the possible mechanisms by immunohistochemistry and Western blotting.Our results showed that TSG treatment rescued the spatial and non-spatial learning and memory impairments of APP/PS1 mice at Morris water maze test and novel object recognition test.Furthermore,Aβ40/42 deposition in the cortex and hippocampus of APP/PS1 mice treated with TSG was significantly reduced compared to the wild type mice using the immunohistochemical technique.Finally,Western blotting showed that TSG primarily decreased the APP expression to avoid the Aβplaque deposition in the cortex and hippocampus of mice.These results reveal the beneficial effects of TSG in APP/PSI-AD mice,which may be associated with the reduction of Aβdeposits in the brain.

Tetrahydroxy stilbene glucoside reduces the cognitive impairment and overexpression of amyloid precursor protein induced by aluminum exposure

Objective Excessive aluminum （Al） exposure impairs neurocognitive function in humans and animals. Epidemiologic studies have shown a potential linkage between chronic Al exposure and Alzheimer’s disease. The present study aims to evaluate the effects of tetrahydroxy stilbene glucoside （TSG）, the extract from herbal medicine Polygoni Multiflori, on cognitive impairment and the over-expression of hippocampal amyloid precursor protein （APP） induced by chronic exposure to Al in rats. Methods Rats were treated with 0.3% aluminum chloride （AlCl3） prepared in the drinking water for 90 d. AlCl3-treated animals were then randomly assigned to receive vehicle, TSG （4 g/kg）, or Vitamin E （VE; 40 mg/kg） treatment for 5 months. VE served as a positive control. The effect of TSG was evaluated by passive avoidance task, and APP expression was evaluated by Western blotting. Results Following exposure to AlCl3 for 90 d, animals displayed a striking decrease （〉80%） in step-through latency in the passive avoidance task and a significant increase in the expression of APP in the hippocampus. Both TSG and VE significantly ameliorated the performance impairment in the passive avoidance task, and suppressed the over-expression of APP. Moreover, the effects of TSG, but not of VE, were in a time-dependent manner. Conclusion TSG may possess therapeutic effects against Alzheimer’s disease.

Tetrahydroxy stilbene glucoside improved the behavioral disorders of APP695V717l transgenic mice by inhibiting the expression of Beclin-1 and LC3-Ⅱ

OBJECTIVE:To observe the effect of tetrahydroxy stilbene glucoside(TSG) on the behavior of APP695V717 I transgenic mouse models and the expression of autophagy-associated proteins Beclin-1and LC3-Ⅱ.METHODS:Forty 3-month-old APP695V717 I transgenic mice were randomized equally into either a TSG group(n = 20) or a model group(n = 20).A normal control group consisted of C57BL/6J mice of the same age and background(n = 20).The TSG group received TSG intragastric administration for1 month.Behavior was measured using the Morris water maze and the Y-maze tests.Changes in protein expression and mRNA of autophagy-associated Beclin-1 and LC3-Ⅱ in mice hippocampus were detected by western blot and Reverse Transcription-Polymerase Chain Reaction(RT-PCR) analyses.RESULTS:The number of electric-stimulus escapes significantly increased and the Morris water maze test showed prolonged escape latency,greater swimming distance,less time taken to cross the exact former platform location in the model group,and increased mRNA and protein expressions of Beclin-1 and LC3-Ⅱ compared with the control group(P < 0.05).The TSG group showed a decrease in the number of electric-stimulus escapes,shorter escape latency and swimming distance,greater time taken to cross the exact former platform location,and decreased mRNA and protein expressions of Beclin-1 and LC3-Ⅱ compared with the model group(P< 0.05).CONCLUSION:These results indicate that tetrahydroxy stilbene glucoside can decrease expressions of Beclin-1 and LC3-Ⅱ in the autophagy pathway.It can attenuate injury to endoplasmic reticulum functions caused by Ab neurotoxicity,improving learning,memorizing,and spatial orientation behavior in mice.

高效液相色谱法同时测定补肾健骨口服液中4种成分的含量

目的:建立高效液相色谱(HPLC)法同时测定补肾健骨口服液中没食子酸、二苯乙烯苷、大黄素-8-O-β-D-葡萄糖苷、大黄素等4种成分含量的方法。方法:采用HPLC法,色谱柱为Shim-pack GIST C_(18)(250mm×4.6mm,5μm);流动相为乙腈-0.1%磷酸溶液,进行梯度洗脱;柱温为30℃;进样量为10μL;检测波长为285 nm。结果:4种成分线性关系良好,没食子酸、二苯乙烯苷、大黄素-8-O-β-D-葡萄糖苷、大黄素的线性质量范围分别是0.036 970~0.735 800μg、0.205 400~4.108 000μg、0.014 130~0.282 600μg、0.004 853~0.097 060μg;线性方程分别是Y=2 513.700X-18.223(r=0.998 7),Y=2 165.500X+40.317(r=0.999 8),Y=2 480.400X+0.768(r=0.999 8),Y=3 845.800X+0.778(r=0.999 9)。结论:本试验建立的方法稳定可靠、简便、重复性好、准确度高,可为补肾健骨口服液的含量测定提供参考依据。

高效液相色谱法同时测定苁蓉通便口服液中松果菊苷和二苯乙烯苷的含量

建立同时测定苁蓉通便口服液样品中松果菊苷和二苯乙烯苷含量的HPLC方法。采用Tech Mate C18-ST(4.6 mm×250.0 mm,5µm)色谱柱;利用乙腈-0.2%甲酸溶液进行梯度洗脱;检测波长330 nm;流速1.0 mL/min。结果显示,苁蓉通便口服液中的松果菊苷和二苯乙烯苷与相邻色谱峰分离度良好,松果菊苷和二苯乙烯苷分别在0.1593~0.9558µg和0.3853~4.6235µg范围内与峰面积呈良好的线性关系,平均加样回收率分别为98.28%、98.43%(RSD<2.0%),10批苁蓉通便口服液中2种成分的含量分别为7.70~8.35 mg/mL和8.80~14.70 mg/mL。该方法操作简单、准确,可用于苁蓉通便口服液中松果菊苷和二苯乙烯苷的含量测定。

二苯乙烯苷的稳定性研究

目的 考察影响二苯乙烯苷 (THSG)溶液稳定性的因素 ,并计算室温下其晶体粉末的 t0 .9值。方法 采用HPL C法和 HPL C- MS联用法。结果 以 L ichrospher5 - C1 8(2 5 0 m m× 4 .6 m m,5 μm)为固定相 ,甲醇 -水 (4∶ 6 )为流动相 ,检测波长为 310 nm。 THSG进样量在 0 .14～ 2 .80μg与峰面积线性关系良好 ,自制 THSG纯度高达99.94 % ,p H5 .8时 ,THSG与水分子发生加成反应。强酸促使 THSG水解为葡萄糖和苷元 ,并进一步降解为酚类化合物。强碱性条件下 ,THSG氧化为醌类化合物。在中性及弱碱性水溶液中较稳定 ,30 d内含量无明显变化。THSG晶体粉末室温下放置 2 4 0 d含量无明显变化 ,t0 .9值为 10 98d。

不同炮制工艺对何首乌中成分含量的影响

目的考察炮制工艺和炮制时间对何首乌中大黄素、大黄素甲醚、2,3,5,4′四羟基二苯乙烯2OβD葡萄糖苷(简称二苯乙烯苷)、鞣质含量的影响。方法何首乌分别采用黑豆汁蒸、黑豆汁炖、清蒸、黑豆汁高压蒸和黑豆汁屉上蒸的工艺炮制12h和32h,采用黑豆汁炖12,24,36,48h。HPLC测定炮制饮片中二苯乙烯苷、蒽醌含量。皮粉法测定鞣质含量。结果不同炮制工艺炮制一定时间,二苯乙烯苷含量生片>黑豆汁高压蒸片>黑豆汁炖片>黑豆汁蒸片>清蒸片>黑豆汁屉上蒸片;总游离蒽醌含量黑豆汁高压蒸片>黑豆汁蒸片>黑豆汁炖片>清蒸片>黑豆汁屉上蒸片>生片;总蒽醌含量生片>黑豆汁高压蒸片>黑豆汁蒸片>黑豆汁炖片>清蒸片>黑豆汁屉上蒸片。采用黑豆汁炖制,随炮制时间的延长,二苯乙烯苷含量逐渐下降,至48h约为生品含量的17%;总游离蒽醌含量先上升后下降,32h>36h>24h>48h>12h>0h;总蒽醌含量和结合蒽醌逐渐下降。鞣质含量随炮制时间的延长逐渐下降。结论应规范何首乌饮片的炮制工艺。

何首乌有效成分二苯乙烯苷的药代动力学研究

目的 建立小鼠和兔血浆中二苯乙烯苷浓度的HPLC测定方法 ,研究何首乌中二苯乙烯苷在小鼠和兔体内的药代动力学行为。方法 用DiamonsilTM C1 8色谱柱 (2 5 0mm× 4 6mm ,5 μm) ,以乙腈 甲醇 1%甲酸 (15∶18∶6 7)为流动相 ,流速 1 0mL·min- 1 ,检测波长 32 0nm。结果 线性范围为 0 4 1～ 42 0 μg·mL- 1 (γ =0 9999) ,最低检测浓度为0 0 5 1μg·mL- 1 。高、中、低 3种不同浓度的平均回收率分别为 97 98%,10 1 7%和 10 4 5 %,日内精密度RSD分别为8 7%,2 9%和 5 5 %。小鼠和兔iv二苯乙烯苷后药代动力学行为均符合二室模型 ,药代动力学参数分别为 :T1 2 α =1 9,2 7min ;T1 2 β =8 3,13 5min ;K2 1 =6 6 ,4 2h- 1 ;K1 2 =3 8,3 0h- 1 ;K1 0 =16 0 ,11 2h- 1 ;Vc=0 0 90 ,0 198L·kg- 1 ;AUC =6 918,4 5 30mg·h·L- 1 ;CL =1 4 45 ,2 2 0 8L·h- 1 ·kg- 1 。小鼠ig给药后二苯乙烯苷在胃肠道内的吸收不规则 ,且血药浓度很低 ,药代动力学行为不符合房室模型。结论 建立了二苯乙烯苷血药浓度的HPLC测定方法 ,阐明了二苯乙烯苷的药代动力学特征。方法的专属性高 ,操作简便 ,结果准确。

液-质联用法测定小鼠体内二苯乙烯苷代谢物

Aim To study the metabolite of stilbene glucoside in the Chinese traditional medicine Polygonum multiflornm in mice and elucidate its chemical structure by liquid chromatography tandem-mass spectrometry. Methods The stilbene glucoside was injected into the tail vein of mice. Blood samples were collected from artery in the eyepit. The methanol-protein-precipitated plasma sample was introduced into the liquid chromatography-tandem mass spectrometer directly. The analytical column was C_ 18 column (250 mm×4.6 mm ID, 5 μm). The mobile phase consisted of acetonitrile-methanol-water-formic acid (15∶18∶66∶1) for ES +, acetonitrile-methanol-water (15∶18∶67) for ES -. The UV detection wavelength was set at 320 nm. The mass ion source type was ESI. HV capillary was 3 kV. The dry gas was nitrogen gas and the flow rate was set at 318 L·h -1 . The ion source temperature was 150 ℃. Results The stilbene glucoside and its metabolite were separated completely under the chromatography condition. The ions at m/z 600 and m/z 605 were detected under positive ion polarity while the ions at m/z 581 and m/z 402 were detected under negative ion polarity. Conclusion It was proposed that the metabolite of stilbene glucoside injected in vein was its glucuronide conjugate.

何首乌中鞣质与二苯乙烯苷不同配比对大鼠肝功能指标的影响

目的研究何首乌提取物鞣质与二苯乙烯苷不同配比对大鼠肝脏生化指标的影响,探讨何首乌肝毒性的可能物质基础。方法 SD大鼠120只,随机分为4组,每组30只。空白对照组(A组)给予1ml/100g的蒸馏水,B组给予鞣质4.4g.ml-1.kg-1,C组给予鞣质(4.4g/kg)与二苯乙烯苷(1.0g/kg)1∶1配比的药物,D组给予鞣质(4.4g/kg)与二苯乙烯苷(1.0g/kg)2∶1配比的药物。各组连续给药90d,恢复15d。分别于给药60d、90d及恢复期15d时颈总动脉取血,生化法检测肝功能相关指标。结果与A组比较,给药60d,各给药组ALT升高,差异具有统计学意义(P<0.01),B组CHE升高及TBIL降低,差异具有统计学意义(P<0.01),B组和C组TP升高,差异具有统计学意义(P<0.01)。与A组比较,给药90d,B组CHE、TP升高和LDH降低,差异具有统计学意义(P<0.01),C组和D组CHE、LDH、ALB降低,差异具有统计学意义(P<0.01)。与A组比较,恢复期15d,B组TP和ALB升高、C组CHE和ALP降低、D组CHE和LDH降低,差异具有统计学意义(P<0.01)。结论长期大剂量灌胃何首乌中提取的鞣质与二苯乙烯苷不同配比、对大鼠肝脏有损害,但鞣质对肝脏的损害在停药后可以恢复。

二苯乙烯苷对局灶性脑缺血大鼠脑组织细胞凋亡的影响

目的观察何首乌中提取的二苯乙烯苷对局灶性脑缺血再灌注损伤大鼠模型脑组织细胞凋亡及其凋亡调节因子的影响,阐明二苯乙烯苷抗缺血性脑损伤的作用机制。方法采用插线法阻断大脑中动脉,制备局灶性脑缺血再灌注损伤大鼠模型。用流式细胞术PI单染法检测脑组织细胞凋亡百分率,用流式细胞术间接荧光法检测细胞凋亡调节因子Bcl-2和Bax蛋白的表达。结果二苯乙烯苷应用于缺血再灌注损伤大鼠可明显降低脑组织细胞凋亡百分率,升高脑组织Bcl-2蛋白表达,减少Bax蛋白表达,提升Bcl-2/Bax比值。结论二苯乙烯苷具有抑制大鼠局灶性脑缺血再灌注损伤后脑组织细胞凋亡的作用,其作用机制可能与升高抑制凋亡的Bcl-2蛋白表达,抑制诱导凋亡的Bax蛋白表达,降低二者比值有关。

何首乌有效成分二苯乙烯苷的药理活性研究进展

二苯乙烯苷作为何首乌的有效成分之一,具有广泛的生物活性。文章综述了二苯乙烯苷抗衰老、抗动脉粥样硬化、抗高血脂、抗肿瘤、抗炎、清除自由基、保肝等方面的生物活性及可能作用机理,并探讨了其应用前景,为何首乌及二苯乙烯苷的深入研究与临床应用提供科学依据。

二苯乙烯苷β-环糊精包合物的制备及其稳定性研究

目的考察β-环糊精对二苯乙烯苷的稳定作用。方法用水溶液法制备二苯乙烯苷-β-环糊精包合物,采用热重-差示量热分析和紫外光谱法进行确认,采用高效液相色谱(HPLC)法对包合态和游离态的二苯乙烯苷在pH=1.2,4.9,6.8及光照条件下的稳定性进行了研究。结果二苯乙烯苷可与β-环糊精形成包合比为1∶1的包合物,包合态和游离态的二苯乙烯苷在pH=1.2的条件下均较稳定,而在pH=4.9、6.8及光照条件下,包合态的二苯乙烯苷的稳定性均明显提高。结论β-环糊精的包合作用可以提高二苯乙烯苷化学稳定性。

何首乌生长过程中二苯乙烯苷的含量变化

对1～4年生何首乌的主要药用成分二苯乙烯苷含量在不同时期、不同器官中的变化进行测定分析,以明确何首乌的最佳采收时间和采收部位.结果显示:何首乌各器官中块根的二苯乙烯苷含量最高,为5.35%,其次为老根和老茎,营养器官的幼嫩部分及生殖器官中的含量都很低;随着生长年限的增加,块根中二苯乙烯苷含量逐渐增加.块根中二苯乙烯苷的含量从6月至10月逐渐升高,10月含量达到最高,为7.09%,然后含量逐渐降低.何首乌块根各部分中二苯乙烯苷的含量差异较大,周皮含量最高,为9.64%,薄壁组织最低.从二苯乙烯苷的含量变化考虑,10月采收何首乌较为合理,且老根和老茎应进行综合利用.

不同地区制何首乌中二苯乙烯苷含量测定及稳定性考察

目的 :测定不同地区制何首乌中 2 ,3,5 ,4′ 四羟基二苯乙烯 2 O β D 葡萄糖苷 (简称二苯乙烯苷 )含量 ,同时考察其稳定性。方法 :采用高效液相色谱法测定二苯乙烯苷含量。稳定性实验采用高温试验及60 Co γ射线辐射试验。结果 :制何首乌中二苯乙烯苷含量为 0 .12 7%～ 4 .15 0 %。温度及湿度影响二苯乙烯苷稳定性 ,干燥条件下60 Co γ射线辐射稳定。结论

何首乌中二苯乙烯苷的醇提工艺

探索了从何首乌中提取二苯乙烯苷的工艺流程。在比较不同提取方法的基础上 ,确定选用乙醇为提取溶剂。通过单因素和中心组合实验 ,对影响何首乌中二苯乙烯苷提取的因素进行了探讨和研究 ,运用响应面分析法确定了提取工艺的最优化条件 :料液比 (g∶mL)为 1∶9,提取时间为66min ,乙醇体积分数为 61 1 % ,提取次数为 2次。在此条件下 ,二苯乙烯苷的得率可达 6 3 9%。

二苯乙烯苷及其β-环糊精包合物在大鼠体内药代动力学研究

[目的]研究二苯乙烯苷(THSG)单体及其β-环糊精包合物大鼠灌胃后药代动力学行为。[方法]SD大鼠分别灌胃THSG单体及其β-环糊精包合物,HPLC-UV测定血浆样品中THSG含量,采用DAS1.0软件计算主要药代动力学参数[。结果]测定方法准确可靠,符合生物样品测定要求。THSG单体及其β-环糊精包合物的Cmax分别为(0.24±0.09)、(1.16±0.80)mg/L;AUC分别为(11.50±3.97)、(32.24±7.77)μg/(L.min),经t检验两者差异有统计学意义(P<0.05)[。结论]β-环糊精的包合作用可通过促进THSG的吸收及减缓其消除,有效的提高THSG的体内生物利用度。

何首乌及其炮制品中二苯乙烯甙含量测定

应用一阶导数光谱法测定何首乌及其炮制品中二苯乙烯甙的含量。实验结果表明:本法操作简便,重现性好,结果稳定,可用为何首乌炮制质量标准之一。

Aβ诱发细胞钙失衡中蛋白激酶A的表达变化及二苯乙烯苷的影响

目的探讨β-淀粉样肽(Aβ)诱发的细胞内钙失衡机制中蛋白激酶A(PKA)的作用及二苯乙烯苷的干预效果。方法在立体定向仪下大鼠海马部位注射Aβ1-42造AD模型,大鼠随机分为对照组、手术组、模型组、二苯乙烯苷(TSG)组;免疫荧光法检测的胞内钙离子,在激光扫描共聚焦显微镜观察;磷基转移法测定细胞膜和胞浆的PKA活性,免疫印迹法检测PKA蛋白表达。结果与对照组比较,模型大鼠细胞内钙离子浓度增高;PKA活性及表达均明显增高,差异有显著性;与模型组相比,二苯乙烯苷干预后,细胞内钙离子降低,PKA活性及表达均下降,差异有显著性。结论 Aβ神经毒性作用引起的细胞内钙失衡机制涉及PKA活化后促进钙内流,并以细胞膜相活化更明显;二苯乙烯苷通过抑制PKA活化及表达,以纠正细胞内的钙超载。