飞龙掌血提取物的镇痛作用及相关机制研究

  目的：对飞龙掌血的镇痛作用及相关机制进行实验探讨,为进一步研究开发镇痛中药提供参考  方法：采用醋酸扭体法和热板法考察飞龙掌血的提取物对于化学刺激和热刺激引起疼痛的镇痛效果;用酶联免疫法(ELISA法)检测小鼠血清中β内啡肽(β-EP)、前列腺素E₂(PGE₂)、五羟色胺(5-HT)、一氧化氮(NO)的含量;用Western blotting法检测小鼠脑组织中β-EP、PGE₂和阿片μ受体的蛋白表达  结果：飞龙掌血提取物能明显减少冰醋酸所致小鼠的扭体次数,且醇提物低于水提物(P<0.01);飞龙掌血提取物能提高小鼠热刺激痛阈值,醇提物与水提物之间无显著性差异(P>0.05);飞龙掌血提取物能增加小鼠血清中的β-EP的含量、降低PGE₂及NO的含量,且醇提物更明显(P<0.05),而5-HT的含量无变化(P>0.05);飞龙掌血的提取物还能上调小鼠脑组织中β-EP的蛋白,下调PGE₂的的蛋白表达(P<0.05)  结论：飞龙掌血具有镇痛作用,且醇提物效果优于水提物,高剂量组优于中、低剂量组,其镇痛机制可能是与通过增加血清中β-EP,降低PGE₂、NO的含量,上调脑组织中β-EP的蛋白表达、下调PGE₂的表达有关。     

五味子醇甲对记忆障碍模型小鼠学习记忆的影响

 目的 观察五味子醇甲对记忆障碍模型小鼠学习记忆的改善作用并探讨其机制。方法 在整体水平,五味子醇甲（0.5、1.0、2.0 g·kg-1·d-1）连续灌胃14 d后,以戊巴比妥钠（20 mg·kg-1）建立小鼠记忆障碍模型,采用Morris水迷宫法检测小鼠学习记忆能力;测定脑组织超氧化物歧化酶（superoxide dismutase,SOD）、一氧化氮（nitric oxide,NO）、过氧化氢酶（catalase, CAT）的活性;免疫组化分析核转录因子-κB（NF-κB）、乙酰胆碱转移酶（choline acetyltransferase, ChAT）、脑源性神经营养因子（brain-derived neurotrophic factor ,BDNF）的表达。在细胞水平,以PC12为靶细胞,以过氧化氢（H₂O₂）诱导细胞凋亡,以五味子醇甲（0.3,0.6,1.2 μmol·L-1）进行预保护24 h,检测PC12细胞活性及细胞上清液中NO的变化;细胞免疫化学染色检测PC12细胞中bcl-2的表达;Western blot方法检测bax的表达。结果 与模型组比较,五味子醇甲可延长小鼠有效区停留时间、增加小鼠穿越平台的次数;提高小鼠脑内CAT的活性、SOD活性并降低NO含量（P＜0.05,P＜0.01）。免疫组化结果显示,五味子醇甲可增加海马区ChAT蛋白表达、BDNF的蛋白表达、抑制NF-kB蛋白的核表达,与模型组比较有统计学差异（P＜0.05,P＜0.01）。五味子醇甲可提高PC12细胞的存活率并降低其细胞上清液中NO的浓度,与H₂O₂模型组比较有差异（P＜0.05,P＜0.01）;细胞免疫化学染色提示五味子醇甲可增加bcl-2的表达量,Western blot 结果表明,高剂量组五味子醇甲可降低bax蛋白表达量。结论 五味子醇甲具有改善记忆障碍模型小鼠学习记忆的作用,其机制可能与抑制NF-κB的核转录、增加BDNF的表达而有效降低脂质过氧化产物水平有关;亦可通过增强海马区ChAT的表达,增加ACh递质的合成;或是在细胞水平调节Bcl-2家族蛋白的表达等多种途径有关。     

干姜对缺氧和受寒小鼠的影响

抗缺氧实验表明,干姜醚提物可能是通过减慢机体耗氧速度,延长常压密闭缺氧和KCN中毒缺氧模型小鼠的存活时间,也能延长断头小鼠的张口动作持续时程,但对NaNO₂中毒小鼠存活时间仅有延长倾向。     

吴茱萸提取物乳膏镇痛作用及其机制探讨

 目的 研究吴茱萸提取物乳膏镇痛作用及其机制。方法 以昆明小鼠为模型动物,采用热板法和福尔马林致痛法研究吴茱萸提取物乳膏镇痛作用;采用酶联免疫法测定降钙素基因相关肽及前列腺素E₂的含量,HE染色法考察疼痛部位组织炎性细胞浸润程度,探讨吴茱萸提取物乳膏在体镇痛机制。结果 0.1%~0.4%吴茱萸提取物乳膏能缓解热所致小鼠疼痛（P<0.05）,对福尔马林所致两相疼痛均有明显镇痛作用（P<0.05）,并呈载药量依赖性。酶联免疫法结果表明,0.2%吴茱萸提取物乳膏能明显降低皮下注射福尔马林3 min后疼痛部位皮肤中降钙素基因相关肽表达量（P<0.05）,且降钙素基因相关肽量随时间的延长而减少（P<0.05）;但0.1%~0.4%吴茱萸提取物乳膏对血浆中降钙素基因相关肽和前列腺素E₂含量无显著影响,对皮肤中前列腺素E₂含量无影响。切片结果证实,0.1%~0.4%吴茱萸提取物乳膏能显著降低福尔马林引起的炎症细胞浸润程度（P<0.05）。结论 局部施用吴茱萸提取物乳膏对小鼠有镇痛作用,并呈载药量依赖性,其镇痛机制可能与耗竭并阻断降钙素基因相关肽传导并协同抗炎作用有关。     

灯盏花素对缺血再灌注诱导小鼠学习记忆损伤的保护作用

 目的探讨灯盏花素对脑缺血再灌注诱导小鼠学习记忆损伤的保护机制。方法双侧颈总动脉结扎法建立重复前脑缺血再灌注模型。小鼠随机分为假手术组、模型组、阳性药物对照组(银杏叶提取物,35mg·kg^-1,ip)和灯盏花素治疗组(80,20,5mg·kg^-1,ip)。小鼠适应3d后,灯盏花素治疗组和阳性药物对照组分别腹腔注射灯盏花素和银杏叶提取物,连续给药7d。模型组和对照组在相同时间点注射等量生理盐水。第7天给药30min后,用1.275g·kg^-1乌拉坦腹腔注射麻醉小鼠,重复缺血再灌注手术。术后第2天用开场行为模型检测小鼠在新异环境中的自发行为;第3天用水迷宫检测小鼠学习记忆能力;用紫外分光光度计检测脑组织超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量;同时检测灯盏花素对H₂O₂引起红细胞溶血的影响。结果前脑重复缺血再灌注并没有明显影响小鼠在新异环境中的自发活动和探究行为,但使小鼠的学习记忆能力显著下降,同时伴随脑内SOD活性降低和MDA含量升高。5,20和80mg.kg^-1灯盏花素及银杏提取物能显著提高缺血小鼠学习记忆能力(P<0.05,P<0.01),并提高脑组织SOD活性(P<0.01),使MDA含量明显下降(P<0.05,P<0.01);体外实验表明,0.015g.L^-1以上剂量的灯盏花素能明显降低H₂O₂引起的红细胞溶血率(P<0.05,P<0.01),且呈剂量依赖关系。结论灯盏花素能减轻缺血再灌注引起的脑损伤,改善小鼠学习记忆能力,该作用可能与其提高脑组织抗氧化能力有关。     

乌蕨对砷和铵的解毒作用

观察了乌蕨的4种提取物对小鼠砷和氯化铵急性中毒的解毒作用。结果表明:乌蕨提取物B及C能明显降低小鼠的死亡率,提取物C尚可使小鼠对砷耐受量提高,使其LD₅₀从31.1±4.3mg/kg提高到38.2±5.9mg/kg。4种提取物对小鼠NH₄Cl中毒虽可降低其死亡率,但与对照组相比,差异无显著意义(P>0.05)。     

玛咖不同提取物和单体对小鼠脾淋巴细胞增殖及诱生IL-2和TNF-α的影响

目的 通过观察玛咖不同提取物和单体对小鼠的免疫调节作用,探讨其对小鼠脾淋巴细胞增殖及诱生白细胞介素-2（IL-2）和肿瘤坏死因子-α（TNF-α）的影响。方法 用反相硅胶柱（ODS柱）将玛咖甲醇提取物进行分段富集,获得Fr₁, Fr₂, Fr₃, Fr₄, Fr₅和Fr₆ 6个部位及3个单体。设置玛咖不同提取物和单体的不同给药剂量组,采用细胞增殖与活性检测法（CCK-8）检测其对无丝裂原组、刀豆蛋白A（Con A）诱导组、脂多糖（LPS）诱导组模型小鼠脾淋巴细胞增殖的影响,采用酶联免疫吸附测定试验法（ELISA）测定IL-2和TNF-α的含量。结果 与模型组比较,玛咖提取物Fr₃,Fr₆,单体N-苄基十六碳酰胺和1,2-二氢-4-甲醛-3-苯甲基-N-羟基吡啶对Con A诱导的T淋巴细胞和LPS诱导的B淋巴细胞存在一定促进增殖作用（P<0.01）,增殖作用较弱,玛咖提取物和单体能显著诱导脾淋巴细胞分泌IL-2和TNF-α（P<0.01）。结论 玛咖提取物及单体能够通过增强IL-2和TNF-α的分泌发挥免疫增强作用,同时可以一定程度上促进脾淋巴细胞的增殖。作用药效物质基础推测可能是玛咖酰胺和吡啶类生物碱,具体作用机制仍需进一步探讨。     

龙胆对小鼠急性肝损伤保护作用的研究

目的:目的:研究龙胆地上部分和根提取物对小鼠急性肝损伤的保护作用。方法:以CCl₄、硫代乙酰胺(TAA)和D-半乳糖胺(D-GlanN)分别给小鼠灌胃造肝损伤模型,观察龙胆地上部分提取物和龙胆根提取物对模型小鼠血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)和碱性磷酸酶(AKP)水平的影响。结果:龙胆地上部分提取物各剂量均可显著降低CCl₄和TAA急性肝损伤小鼠血清ALT,AST和AKP水平(P<0.05),而对D—GlanN模型小鼠无明显作用(P>0.05),并且实验结果与龙胆根提取物作用一致。结论:龙胆地上部分和根提取物对小鼠急性肝损伤均有一定的保护作用,因此,龙胆地上部分可代替根使用。     

安宫牛黄丸及其简化方的药效学比较研究

目的:根据安宫牛黄丸的治疗学作用,比较研究安宫牛黄丸及其简化方(安宫牛黄丸全方仅缺朱砂和雄黄)的药效学作用,以探讨安宫牛黄丸中朱砂和雄黄对其药效作用的影响。方法:实验以伤寒菌苗致家兔发热、戊巴比妥钠诱导小鼠睡眠、NaNO₂诱导小鼠缺氧死亡、硝酸士的宁及戊四唑诱发小鼠惊厥为模型,观察比较安宫牛黄丸及其简化方的药效学作用。结果:安宫牛黄丸及其简化方有明显的解热作用;安宫牛黄丸及其简化方与戊巴比妥钠有明显的协同镇静作用;对NaNO₂诱导的小鼠缺氧死亡有明显的保护作用;安宫牛黄丸及其简化方对硝酸士的宁及成四唑诱发小鼠惊厥无明显的保护作用。结论:安宫牛黄丸及其简化方在上述药效学研究中,两者在药效学上无明显差异。     

马勃孢子粉醇提物对小鼠全层皮肤缺损性创面的治疗作用考察

目的 比较马勃孢子粉和子实体的不同溶剂提取物对成纤维细胞和小鼠全层皮肤缺损性创面的药效差异，筛选马勃治疗创面的最佳用药部位。方法 将马勃孢子粉和子实体的水提物、醇提物对体外培养的小鼠皮肤成纤维细胞（MSF）进行干预，通过细胞增殖与活性检测（CCK-8）法比较细胞增殖、划痕实验比较细胞迁移、流式细胞技术探究细胞周期、酶联免疫吸附测定法（ELISA）测定Ⅰ型和Ⅲ型胶原蛋白生成量，同时建立小鼠背部全层皮肤缺损性创面模型，考察马勃孢子粉不同溶剂提取物的治疗效果，并采用紫外-可见分光光度法测定其不同溶剂提取物中指标成分的含量。结果 马勃水提物具有一定细胞增殖抑制作用，醇提物具有促进MSF增殖、迁移、分泌Ⅰ型和Ⅲ型胶原蛋白的作用，且孢子粉功效显著优于子实体。在质量浓度为10 mg·L^-1时，马勃孢子粉醇提物处理的细胞增殖率最高可达（159.22±15.95）%，并促进MSF从G₀/G₁期的静止状态进入S期和G₂/M期的增殖状态，增殖指数增大，增高成纤维细胞迁移率及Ⅰ型、Ⅲ型胶原蛋白的生成量。马勃孢子粉醇提物能有效促进创面愈合、促进创面组织上皮化完整、减少炎性细胞浸润、生成新生胶原纤维。成分含量测定结果显示，与子实体醇提物比较，孢子粉醇提物中多酚及黄酮类成分含量更高。结论 马勃孢子粉醇提物是马勃促进创面愈合的主要活性部位，具有较好的开发前景，其药效成分可能为多酚和黄酮类。     

Improvement of memory in mice by extracts from leaves of Ginkgo biloba L.]

The study has shown that the extracts from leaves of Ginkgo biloba can significantly improve the NaNO2 and scopolamine induced impaired memory in mice. The potency of the ethanolic extract is greater than that of the aqueous extract. The ethanolic extract acts favorably on the memory function of normal animals. Both extracts help to prolong the survival time of mice receiving 200 mg/kg(ip)NaNO2.     

Anxiolytic effect of Rubus brasilensis in rats and mice

The aim of the present work was to investigate if infuse and ethanolic extracts (aqueous, butanolic and wax fractions) of Rubus brasiliensis Martius (Rosaceae) induce anxiolytic effect. The extracts were administered to male Wistar rats and Swiss mice per oral route, at 50, 100 and 150 mg/kg, 30 min before the behavioral evaluation in the elevated plus maze (EPM). Both infuse and wax ethanolic fraction at the dosage 150 mg/kg, vo, increased the number and the percentage of open arm entries of rats and mice. The aqueous and butanolic fractions, obtained from ethanolic extract, failed to induce anxiolytic effect. The treatment of mice with flumazenil (Ro 15–1788), 1.5, 2.0 and 2.5 mg/kg, i.p., 15-min before the administration of infuse or wax fraction, 150 mg/kg, vo, blocked the infuse or wax fraction-induced anxiolytic effect. The LD₅₀ for the wax fraction was 1000 mg/kg. In conclusion, the infuse and wax ethanolic fraction of R. brasiliensis present anxiolytic effect in rats and mice. In addition, it is suggested that the anxiolytic effect may be attributed at least to one liposoluble principle with low acute toxicity which may be acting as an agonist on GABA_A-benzodiazepine receptor complex.     

桂莪术乙醇提取物及其挥发油对实验性肝损害的保护作用

桂莪术醇提物及挥发油对CCI₄、TAA引起小鼠SGPT升高有明显的降低作用,使BSP潴留量减少,相应肝组织病变减轻;对CCI₄损害和正常小鼠戊巴比妥钠睡眠时间明显缩短,对甘油三酯有抑制作用。桂莪术的毒性较低,口服LD₅₀醇提物为86.8±12g生药/kg,挥发油为1.10±0.08g/kg。     

Terminalia arjuna,an ayurvedic cardiotonic,increases contractile force of rat isolated atria

The direct effects of aqueous, ethanolic, chloroform and petroleum ether extracts of Terminalia arjuna (family: Combretaceae) bark, an ayurvedic remedy for cardiovascular disorders, were studied on isolated atria of rats. The aqueous extract produced a substantial positive inotropic effect (EC₅₀ = 0.25 mg/mL) but no change in the rate. The ethanol, chloroform and petroleum ether extracts all decreased the rate of contraction. A slight increase in force was seen with ethanol and chloroform extracts. Higher concentrations of the chloroform extract had a negative inotropic effect. The ethanol extract decreased the maximum following frequency of the left atria. It is concluded that Terminalia arjuna has a significant positive inotropic property on rat atria in vitro which could contribute to its efficacy in treating cardiovascular disorders.     

Effect of crocus sativus L. (saffron) stigma and its constituents,crocin and safranal,on morphine withdrawal syndrome in mice

Crocus sativus L. has been shown to interact with the opioid system. Thus, the effects of aqueous and ethanolic extracts of stigma and its constituents were evaluated on morphine‐withdrawal syndrome in mice. Dependence was induced using subcutaneous (s.c.) injections of morphine for 3 days. On day 4, morphine was injected 0.5 h prior the interaperitoneal (i.p.) injections of the extracts, crocin, safranal, clonidine (0.3 mg/kg) or normal saline. Naloxone was injected (5 mg/kg i.p.) 2 h after the final dose of morphine and the number of episodes of jumping during 30 mm was considered as the intensity of the withdrawal syndrome. Clonidine, the aqueous and ethanolic extracts of saffron reduced the jumping activity. Safranal was injected (s.c.) 30 mm prior and 1 and 2 h after the injection of morphine. It potentiated some signs of withdrawal syndrome. The aqueous extract decreased the movement in all of the doses (80, 160, 320 mg/kg) and the ethanolic extract decreased it in the dose of 800 mg/kg in open field test. But crocin and the dose of 400 mg/kg ethanolic extract showed no effect on activity in this test. It is concluded that the extracts and crocin may have interaction with the opioid system to reduce withdrawal syndrome. Copyright © 2009 John Wiley & Sons, Ltd.     

Protective effect of fruits of Morinda citrifolia L. on scopolamine induced memory impairment in mice: a behavioral, biochemical and cerebral blood flow study

Ethnopharmacological relsevance

Noni (Morinda citrifolia L.) is widely used for different illnesses including CNS disorders. Recently Noni has been reported to prevent amyloid beta induced memory impairment in mice. However, the influence of Noni on cholinergic system has not been explored so far. Therefore, present study was designed to investigate effect of Noni fruit on memory, cerebral blood flow (CBF), oxidative stress and acetylcholinesterase (AChE) activity in scopolamine induced amnesia model.

Materials and methods

Mice were orally treated with ethanolic extract of Noni fruit and chloroform, ethyl acetate and butanol fractions of ethanolic extract for three days. Scopolamine was administered 5 min prior to acquisition trial and memory function was evaluated by passive avoidance test. CBF was measured by laser doppler flowmetry. AChE activity and oxidative stress parameters were estimated in mice brain at the end of behavioral studies. Further, effect of ethanolic extract and its fractions (5-400 μg/ml) on AChE activity was measured in vitro.

Results

Scopolamine caused memory impairment along with reduced CBF, increased AChE activity and oxidative stress in mice brain. Ethanolic extract of Noni fruits and its chloroform and ethyl acetate fractions significantly improved memory and CBF. However, butanol fraction had no effect. Further, increased oxidative stress and AChE activity following scopolamine was significantly attenuated by ethanolic extract of Noni and its fractions. Moreover ethanolic extract and its fractions showed dose dependent inhibition of AChE activity in vitro.

Conclusion

These observations suggest that Noni may be useful in memory impairment due to its effect on CBF, AChE and oxidative stress.     

九节木不同极性提取物对小鼠学习记忆能力的影响

目的：探讨九节木不同极性提取物对小鼠学习记忆能力的影响。方法：采用95%乙醇进行提取,然后依次用石油醚、乙酸乙酯、正丁醇进行萃取,得到各部分提取物,各种提取物灌胃给药的剂量为200 mg·kg^-1,脑复康组的给药剂量为120 mg·kg^-1。运用跳台法检测九节木不同萃取物对盐酸氯丙嗪、亚硝酸钠、乙醇、东莨菪碱所致记忆障碍小鼠学习记忆能力的影响,并筛选出活性部位。结果：九节木的提取物能延长小鼠跳台实验的潜伏期,减少错误次数,活性部位为乙酸乙酯部位（P<0.05）。结论：九节木的乙酸乙酯提取物和水提取物具有改善学习记忆障碍小鼠学习记忆能力的作用,其中乙酸乙酯部位为活性部位,且其作用呈现一定的量效关系。     

Antioxidant Activity of Moringa oleifera Tissue Extracts

Moringa oleifera is an important source of antioxidants, tools in nutritional biochemistry that could be beneficial for human health; the leaves and flowers are used by the population with great nutritional importance. This work investigates the antioxidant activity of M. oleifera ethanolic (E1) and saline (E2) extracts from flowers (a), inflorescence rachis (b), seeds (c), leaf tissue (d), leaf rachis (e) and fundamental tissues of stem (f). The radical scavenging capacity (RSC) of extracts was determined using dot‐blots on thin layer chromatography stained with a 0.4 mm 1,1‐diphenyl‐2‐picrylhydrazyl radical (DPPH) solution; spectrophotometric assays were recorded (515 nm). Antioxidant components were detected in all E1 and E2 from a, b and d. The best RSC was obtained with E1d; the antioxidants present in E2 reacted very slowly with DPPH. The chromatogram revealed by diphenylborinate‐2‐ethylamine methanolic solution showed that the ethanolic extract from the flowers, inflorescence rachis, fundamental tissue of stem and leaf tissue contained at least three flavonoids; the saline extract from the flowers and leaf tissue revealed at least two flavonoids. In conclusion, M. oleifera ethanolic and saline extracts contain antioxidants that support the use of the plant tissues as food sources. Copyright © 2012 John Wiley & Sons, Ltd.     

Green tea extract weakens the antibacterial effect of amoxicillin in methicillin‐resistant Staphylococcus aureus infected mice

Tea (Camellia sinensis) has been known for its modulation of resistance of methicillin‐resistant Staphylococcus aureus (MRSA) to β‐lactam antibiotics in vitro. This study aimed to confirm the in vitro effect of green tea extracts with β‐lactams and to determine whether green tea extracts can reduce the minimum inhibitory concentrations (MICs) of amoxicillin in MRSA‐infected mice. The catechins in the test tea that account for the reduced resistance to β‐lactams were quantitatively determined by high‐performance liquid chromatography. The MICs of the ampicillin, cefazolin, amoxicillin, oxacillin, tea extract alone and tea extract in combination with β‐lactams were determined. Proportions of tea extracts and amoxicillin‐tea extract combinations were administered to groups of mice enterally. The in vitro experiment showed that the MICs of four β‐lactams were greatly decreased in the presence of 0.25% tea extract. However, in an in vivo experiment, amoxicillin in combination with 5% tea extract conferred a higher ED₅₀ than that of antibiotic alone. Green tea extract, alone or in combination with amoxicillin, does not have protective benefits in MRSA‐infected mice. This study concluded that tea extract weakened the antibacterial effect of amoxicillin in MRSA infected mice. Tea drinking is not recommended in combination with amoxicillin treatment. Copyright © 2009 John Wiley & Sons, Ltd.     

Terpenoid content of Valeriana wallichii extracts and antidepressant‐like response profiles

Three extracts of Valeriana wallichii DC (Valerianaceae) rhizome and fluoxetine were studied for antidepressant‐like activity in two behavioral models, namely the forced swim test (FST) and the tail suspension test (TST). Fluoxetine as well as methanolic and aqueous extracts of V. wallichii induced monophasic dose‐related decrements in immobility times in both tests. However, the aqueous‐ethanolic fraction induced a biphasic dose‐response profile since it produced a graded effect up to 200 mg/kg but the highest dose (250 mg/kg) was inactive in the FST. This extract also exhibited significantly reduced activity at 200 mg/kg compared to lower doses in the TST. The highest doses of aqueous‐ethanolic extract also reduced locomotor activity which will have led to a negative functional interaction with antidepressant‐like effects. Qualitative phytochemical analysis revealed that the aqueous‐ethanolic extract of V. wallichii was the only separated rhizome fraction containing terpenoids. Furthermore, since the methanolic and aqueous extracts were active in the tests, it is suggested that the antidepressant‐like action of this herbal plant is not contingent upon its terpenoid constituents. Copyright © 2009 John Wiley & Sons, Ltd.