Bioactivity guided isolation of mosquito larvicide from Piper longum

Objective:To isolate the larvicidal component from the fruits of Piper longum（P.longum） against the filariasis vector,Culex quinquefasciatus（C.quinquefasciatus）.Methods:Pulverized fruits of P.longum were subjected to soxhlet extraction using series of organic solvents of increasing polarity.All the solvent extracts were verified for their larvicidal efficacy against 4th instar larvae of C.quinquefasciatus employing standard WHO procedure.Bioassayguided fractionation through column chromatography lead to the isolation of a bioactive amide, pipyahyine from the petroleum ether extract.Results:Petroleum ether extract was found to be the most active fraction among all the extracts tested with LC50 and LC90 being 1.03 and 2.04 ppm respectively.Whereas,pipyahyine,an isolated component of the same fraction was found to be even more effective than the parent extract in terms of LC50 being 0.58 and 1.88 ppm respectively. Conclusions:From the results,it is evident that P.longum can be considered as a powerful arsenal for the control of mosquito population.

荜茇提取物对CCl_(4)诱导大鼠肝纤维化的影响

目的探究荜茇提取物对四氯化碳(CCl_(4))诱导大鼠肝纤维化的作用机制。方法将60只SD大鼠随机分成空白组、模型组、荜茇提取物低剂量组(16.6mg/kg)、荜茇提取物中剂量组(33.3mg/kg)和荜茇提取物高剂量组(66.7mg/kg),每组各12只。除空白组外,其余各组均给予皮下注射50%CCl_(4)-菜籽油溶液干预8周,建立肝纤维化模型。造模的同时给予模型组灌胃0.9%氯化钠溶液,荜茇提取物低、中、高剂量组连续灌胃治疗8周。实验结束后,测定大鼠血清谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)、谷氨酰基转移酶(γ-glutamyltransferase,γ-GT),肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6);苏木精-伊红染色(hematoxylin-eosin staining,HE)、Masson染色观察大鼠肝组织病理变化。提取RNA并采用qPCR技术检测大鼠肝组织中Ⅰ型胶原蛋白(CollagenⅠ)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、TNF-αmRNA表达水平;提取蛋白采用Western blot法检测大鼠肝组织中CollagenⅠ、α-SMA、TNF-α蛋白表达水平。结果HE染色和Masson染色结果可观察到模型组大鼠肝脏纤维增生明显,大鼠血清中ALT、AST、γ-GT升高,血清中炎性介质TNF-α和IL-6分泌增多,CollagenⅠ、α-SMA和TNF-αmRNA和蛋白表达增高,荜茇提取物可明显改善肝纤维化程度,血清中ALT、AST、γ-GT下降,血清中炎性介质TNF-α和IL-6下降,肝组织中CollagenⅠ、α-SMA、TNF-αmRNA和蛋白水平明显下降。结论荜茇提取物对CCl_(4)诱导的大鼠肝纤维化有明显治疗作用,可通过抑制炎性细胞因子TNF-α和IL-6表达,可能通过抑制肝星状细胞激活,从而减少CollagenⅠ和α-SMA合成发挥抗纤维化的药效。

New amide alkaloids from Piper longum fruits

Three new amide alkaloids piperlongumamides A-C(1-3),together with 12 known ones(4-15),were isolated from the fruits of Piper longum.The structures of the new isolates were determined using spectroscopic data analyses.Cytotoxic activity of these amides against HL-60(human leukemia),A-549(human lung cancer),MCF-7(human breast cancer),SMMC-7721(human liver cancer)and SW480(human rectal cancer)cell lines were evaluated.Piperchabamide B(11)exhibited weak inhibitory activity against HL-60(IC_(50)=21.32μM),A-549(IC_(50)=23.82μM)and MCF-7(IC_(50)=16.58μM)cell lines.

Overview of chemical constituents and pharmacological activities of Piper longum Linn.

Piper longum Linn. is a native of the Indo-Malaya region, belongs to the family Piperaceae. It is a slender aromatic climber widely distributed in the tropical and subtropical regions. As a traditional ethnic drug, P. longum contains various chemical constituents such as alkaloids, volatile oil and lignans. The extract of P. longum has been reported to show various activities like antitumor, antioxidant and hepatoprotective activity. Hence, this paper is presented as a review centered on the main chemical constituents and pharmacological effects of P. longum, aiming at its further development and potential value.

Network pharmacology-based prediction and verification of the molecular targets and pathways for Piper longum Linn,against liver cancer

Liver cancer is a common malignant tumor,which is one of the most common causes of cancer related deaths worldwide,specially,in China.Piper longum Linn,is a traditional Chinese medicine that is used to treat and prevent diseases in clinical practice,however,the active ingredients and potential targets of its action against liver cancer remain unclear.In this study,network pharmacology approach was used to identify active ingredients and the major targets of Piper longum against liver cancer.In addition,the key pathways were also predicted on the network pharmacology level,which might be beneficial to the chemical basis and pharmacological studies of this plant.

Biotechnological intervention in Betelvine(Piper betle L.):A review on recent advances and future prospects

Betelvine(Piper betle L.) is cultivated for its deep green heart shaped leaf for(15-20) million Indian and 2 billion foreign consumers annually. The crop provides Rs(6 000-7 000) million of national income per year and at the same time leaves worth Rs(30-40) million is exported to other countries. The leaves are not only used directly for chewing purposes but also possesses antioxidant, anti-inflammatory, anti-apoptotic, anti-cancer and anti-microbial properties. Besides, the leaves also contain eugenol rich essential oil(1%-3%) which is the source for medicine, stimulant, antiseptic, tonic and other ayurvedic formulations. The essential oil also contains chavibetol, caryophyllene and methyl eugenol which are the potent source for preparation in ayurvedic medicine and herbal products. Cost of betelvine essential oil is 10$ per 5 mL. In spite of its great economical and medicinal importance betelvine is still neglected by the researchers for proper characterization and authentication for selection of elite landraces. Lack of awareness among people, use of same planting material for many generations, existing of many synonyms for a single landraces, no proper characterization of available landraces are some of the significant constraints for its commercialization. Our review endeavours a complete advance in the research on betelvine, existing lacunae for its proper characterization and commercial cultivation. It also attempts to provide a comprehensive account on biotechnological interventions made in betelvine aimed at complementing conventional programmes for improvement of this nutraceutically important cash crop.

Quantification of total phenol,flavonoid content and pharmacognostical evaluation including HPTLC fingerprinting for the standardization of Piper nigrum Linn fruits

Objective:To carry out the physicochemical and phytochemical standardization with high performance thin layer chromatography fingerprinting of Piper nigrum L.(P.nigrum)fruits in order to ascertain the standard pharmacognostical parameters of this king of spices.Methods:Many standardization parameters like extractive values,total ash value,water soluble ash value and acid insoluble ash,moisture content,loss on drying and pH values of P.nigrum L.fruits were analyzed.The method of Harborne was adopted for the preliminary phytochemicals screening.Analysis of total phenolic and flavonoid contents,pesticides residues,aflatoxin and heavy metals were also performed.CAMAG-high performance thin layer chromatography system was used for fingerprinting of methanolic extract of P.nigrum L.fruits.Results:The results of phytochemicals testing indicated the presence of carbohydrates,phenolic compounds,flavonoids,alkaloids,proteins,saponins,lipids,sterols and tannins in various solvent extracts.Total phenolic and flavonoid contents in methanolic extract were found to be 1.728 1 mg/g and 1.087 ug/g,respectively.Heavy metals concentrations were found to be within standard limits.Aflatoxins and pesticides residues were absent.Conclusions:The outcome of this study might prove beneficial in herbal industries for identification,purification and standardization of P.nigrum L.fruits.

Fusarium solani Upregulated Sesquiterpene Synthase Expression, Sesquiterpene Production and Allelopathic Activity in Piper betle

Activators of sesquiterpene synthase(STS) gene expression and sesquiterpene production in Piper betle L. were examined using quantitative real time PCR and gas chromatography mass spectrometry methods, and the allelopathic activity of untreated and Fusarium solani-treated betel extracts was tested on seed germination and on the shoot and root growth of Thai rice variety PSL2(Oryza sativa cv. Phitsanulok 2) and three dominant paddy weeds(Eclipta prostrata, Echinochloa crus-galli and Chloris barbata). The results demonstrated that F. solani dramatically upregulated STS expression and productions of β-cubebene, β-caryophyllene and germacrene D sesquiterpene when compared with the untreated control, and that betel extracts had a greater inhibitory effect on weeds than on rice. The effects were more clearly detected on seed germination and root growth than on shoot growth, and they were found to be dose-dependent. It is also noted that F. solani-treated extract had stronger effects than the untreated extract. The species most sensitive to the allelopathic effects was C. barbata, germination of which was completely inhibited even at a dose of 0.1 mg/mL untreated extract. With regards to rice, although betel extract at 1.0 mg/mL showed no inhibition on germination, it affected the elongation of rice roots, in addition to those of the tested weeds. The obtained data suggested that F. solani has potential as an activator of sesquiterpene allelochemical production via STS expression, the latter leading to the treated betel extract having a stronger phytotoxic effect. These results were beneficial in the promotion of natural herbicide production using biotechnology.

Isolation of Bulk Amount of Piperine as Active Pharmaceutical Ingredient (API) from Black Pepper and White Pepper (<i>Piper nigrum</i>L.)

In the pharmaceutical world the majority of the active pharmaceutical ingredients (API) have been obtained from the natural products. Piperine is such naturally occurring alkaloid which can be considered as major bioactive phytochemical having broad spectrum of pharmacological activities. It is obtained from the most valuable ethnomedicinal spices peppercorns i.e. black pepper and white pepper, which are the fruits of the Asian vine Piper nigrum L. Because of the widespread traditional uses of this medicinal compound, present article reveals a simple and effective isolation method of bulk piperine. The novelty of this investigation is to provide an idea for utilizing such natural method of large scale commercial piperine production as API drug in spite of chemical synthesis. Piperine was isolated in a pure crystal form and characterized by its melting point, X-Ray diffraction (XRD) studies and spectral data, including two-dimensional nuclear magnetic resonance (2D-NMR) spectroscopy. Chromatographic techniques like Thin Layer Chromatography (TLC) and High Performance Liquid Chromatography (HPLC) were applied to determine the purity of the yielded piperine. It was found that piperine yield from black pepper was within 2.5% - 3.0% and from white pepper within 4.0% - 4.5% and the purity of the yielded piperine was found to be up to 98.5% for black pepper and 98.2% for white pepper. Considering this yield value and purity it is indicated that, such effective isolation method can be successfully utilized for industrial large-scale production commercially. According to the result, it can be claimed that, as a natural product the isolated piperine can also be utilized as API drug like other expensive chemically synthesized piperine in different drug formulation.

Research progress on the main chemical constituents and pharmacological effects of Piper betle L.

The Piper betle,belonging to Piper genus of the Piperaceae family,is an evergreen and perennial creeper used in several traditional medicines.This plant has been known to possess antioxidant,antitumor,antibacterial,anti-inflammatory and antimalarial activities.A wide range of chemical compounds including alkaloids,simple phenylpropanoids,lignans,flavonoids,steroids,triterpenoids,sesquiterpenes,monoterpenes have been isolated from this plant.The present review summarized the information concerning phytochemistry and biological activity of P.betle,providing references for the future research of this plant.

Progress on chemical constituents and pharmacological effects of Piper nigrum L.

Piper nigrum L.,belonging to Piper genus of the Piperaceae family,is a medicinal and edible plant.Studies have shown that there are many chemical constituents in this plant,including alkaloids,lignans,steroids and other compounds.In addition,some chemical components of P.nigrum have potential values to treat cancer,inflammation and other diseases.Research progress on the chemical constituents and pharmacological effects of P.nigrum was reviewed in this study to better explore its potential medicinal value.

荜茇提取物的抗氧化活性及改善LPS所致大鼠听力损伤的作用研究

目的 探讨荜茇提取物的抗氧化活性及改善LPS所致大鼠听力损伤的作用。方法 测试浓度为0.05~1.0 mg/mL的荜茇提取物对DPPH自由基的清除率;测试浓度为25~500μg/mL荜茇提取物对LPS诱导RAW 264.7细胞分泌NO的抑制作用。随机将24只SD大鼠分成空白对照组、模型组、氢化可的松组和荜茇提取物组,每组6只,空白对照组不造模处理,模型组、氢化可的松组及荜茇提取物组连续鼓室内注射5 mg/mL的LPS 5 d后,模型组生理盐水灌胃,氢化可的松组1.81 mg/kg氢化可的松灌胃,荜茇提取物组按生药量1 g/mL荜茇提取物灌胃,连续灌胃10 d后,测试各组大鼠8、16 kHz ABR反应阈值、耳蜗组织中谷氨酸、MDA、TNF-α、NO因子的表达量,并行耳蜗基底膜铺片观察。结果 荜茇提取物对DPPH自由基清除的IC50为0.505 mg/mL;浓度为25~500μg/mL的荜茇提取物对LPS诱导的RAW264.7细胞过度分泌NO有显著的抑制作用,抑制率的IC50为280.55μg/mL。连续灌胃10 d后,与空白对照组相比,模型组8、16 kHz ABR阈值显著升高,耳蜗组织中谷氨酸、MDA、TNF-α、NO的表达显著升高,SOD含量显著降低;与模型组相比,氢化可的松组和荜茇提取物组8、16 kHz的ABR反应阈值降低(P<0.01),大鼠耳蜗组织谷氨酸、MDA、TNF-α、NO因子表达也显著降低(分别为P<0.05、P<0.01、P<0.001),SOD活力提高(P<0.05)。基底膜铺片结果示,空白组毛细胞无明显缺失,三排外毛细胞纤毛排列整齐,结构完整,无缺失;模型组耳蜗毛细胞大部分明显缺失,并且细胞肿胀明显、结构严重不完整,氢化可的松组及荜茇提取物组耳蜗毛细胞排列较为整齐,毛细胞肿胀缓解。结论 荜茇提取物可能通过抑制耳蜗组织中的谷氨酸、自由基的过度表达以及炎症反应,提高超氧化歧化酶SOD活力,产生良好的改善听力作用。

Network pharmacology-based virtual screening of active alkaloid ingredients from Piper nigrum L.and the potential targets against Parkinson’s disease

Parkinson’s disease(PD)is a common neurodegenerative disease.Despite a great deal of research on this disease,there is still no definitive treatment plan.Literature investigations have shown that Piper nigrum L.has a protective effect on the nerve center.Therefore,network pharmacology was used in this study to investigate the therapeutic potential of P.nigrum for PD.As a result,six active alkaloid ingredients and four potential targets were obtained.This study laid the foundation for finding the lead compounds from P.nigrum to treat PD.

HPLC法同时测定荜茇-大黄中6种成分的含量

目的建立HPLC法同时测定荜茇-大黄药对中胡椒碱、芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚的含量。方法采用Thermo Hypersil BDS C_(18)色谱柱(4.6 mm×250 mm,5μm);以甲醇(A)-0.1%磷酸(B)为流动相,梯度洗脱;体积流量1.0 ml/min;检测波长254 nm;柱温30℃。结果各成分色谱峰分离度良好,胡椒碱、芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚的质量浓度分别在6.02~240.72μg/ml(r=0.9996)、1.85~74.18μg/ml(r=0.9993)、1.79~71.78μg/ml(r=0.9996)、1.59~63.55μg/ml(r=0.9995)、5.42~216.72μg/ml(r=0.9992)、1.62~64.64μg/ml(r=0.9997)内呈良好的线性关系,平均回收率分别为99.61%(RSD=1.50%)、99.48%(RSD=1.11%)、99.10%(RSD=1.31%)、99.72%(RSD=1.29%)、99.88%(RSD=1.25%)、99.06%(RSD=0.97%)。结论该方法准确稳定,重复性好,可用于荜茇-大黄药对的质量控制。

FTIR方法和HPLC指纹图谱鉴别不同产地荜茇

荜茇是药食同源性中药,也是蒙、藏、维医的习用药材,临床用于治疗胃痛、腹泻等,也被用作香料或调味剂,因此其质量控制非常重要。已报道采用高效液相色谱法检测胡椒碱或荜茇宁,该方法缺乏整体性。傅里叶红外光谱法和HPLC指纹图谱联合应用考察4个产地6批次荜茇,基于共有峰率和变异峰率双指标序列分析荜茇的红外光谱,并建立荜茇的HPLC指纹图谱。结果显示两种方法对荜茇的评价结果相互印证,各批次荜茇的化学成分基本相同,质量比较稳定,其中云南产荜茇(S1)与其余5批次荜茇有一定差异。红外光谱共有峰率和变异峰率与HPLC指纹图谱均能从化学成分的角度评价药材的质量,具有整体性和综合性。荜茇的红外光谱由药材中多种化学成分的红外光谱叠加产生,而指纹图谱仅显示在波长254 nm条件下有紫外吸收的化合物,因此红外光谱在分析药材品质中更具有优势。同时考察了胡椒碱和荜茇宁的含量,云南(S1)和海南(S5, S6)较安徽亳州(S2)和广西(S3, S4)低,结果与红外和HPLC指纹图谱的结果有一定的差异,提示中药材的质量评价应更注重整体性。将FTIR和HPLC指纹图谱联合应用于荜茇的质量控制研究,两种方法互为补充,操作简便,样品使用量少,可做为药材快速鉴别的新方法,为荜茇的化学成分、质量控制和开发应用提供研究基础和依据。

超声波辅助亚临界水提取荜茇总黄酮工艺优化及其抗氧化活性评价

采用超声波协同亚临界水技术进行荜茇中黄酮类成分的提取条件研究,并对其不同部位的总黄酮提取物进行抗氧化性对比分析。结果表明,超声波协同亚临界水提取荜茇整株总黄酮最佳工艺条件为超声温度50℃,超声功率240 W,超声时间40 min,料液比1∶30(g/mL),亚临界水提取温度125℃,亚临界水提取时间为40 min,在此工艺条件下荜茇整株总黄酮最高提取量为(30±0.05)mg/g。不同部位总黄酮提取物对DPPH自由基和超氧阴离子自由基清除能力的大小顺序为果穗>叶>根>茎,DPPH自由基清除率和超氧阴离子自由基清除率的IC50值分别为荜茇果穗37.96、304.33μg/mL,荜茇叶44.55、384.36μg/mL,荜茇根47.68、461.54μg/mL,荜茇茎130.60、487.80μg/mL。采用超声波协同亚临界水提取荜茇整株总黄酮的工艺条件合理,提取得到的总黄酮具有良好的抗氧化性。

HPLC测定海南产胡椒中胡椒碱的含量

测定海南不同地区产的胡椒果实及胡椒根和胡椒叶中胡椒碱的含量。运用索氏提取法对胡椒中胡椒碱进行提取,并采用HPLC对胡椒碱的含量进行测定。海南万宁产胡椒果实中胡椒碱的含量为3.17%,胡椒根中胡椒碱含量为0.43%,叶中胡椒碱含量为0.01%,万宁产的胡椒果实中胡椒碱的含量比其它地区的含量高。不同的生长环境对胡椒果实中胡椒碱的含量有一定的影响。胡椒果实中胡椒碱含量高于根部和叶子中胡椒碱含量。

HPLC法测定胡椒根中胡椒碱的含量

目的建立胡椒根药材中胡椒碱的含量测定方法。方法采用反相高效液相色谱法,色谱柱:Luna C18柱(250mm×4.60mm,5μm);流动相:甲醇-水(77∶23);流速:1.0mL·min-1;检测波长为343nm;柱温:35℃。结果胡椒碱在0.164μg～0.984μg范围内,具有良好的线性关系,r=0.9996;平均回收率为98.09%,RSD=2.67%(n=9)。三批胡椒根药材胡椒碱含量为6.67～6.77mg·g-1。结论胡椒根含有胡椒碱,该含量测定法可用于控制胡椒根药材的质量。

超声波-盐析辅助蒸馏提取黑胡椒精油方法优化及活性分析

采用超声波-盐析辅助水蒸气蒸馏法提取黑胡椒(Piper nigrum L.)精油,利用响应面法确定了最佳的液料比、超声时间、NaCl浓度等提取参数。结果表明,超声波辅助蒸馏法提取黑胡椒精油的最佳条件参数为液料比17∶1、超声时间13 min、NaCl浓度2.4%。在此条件下黑胡椒精油的得率为1.79%。对使用此方法获得的黑胡椒精油进行了抗氧化和抑菌活性检测,结果表明提取物具有一定的抗氧化能力,且对ABTS的清除能力远强于对DPPH的清除能力。同时,提取物对金黄色葡萄球菌和青枯假单胞菌均具有较强的生长抑制功能。

荜拔油不皂化物抑制C57BL/6小鼠胆囊结石的形成

目的:观察荜拔油不皂化物对实验性C57BL/6小鼠胆囊结石形成的抑制作用。方法:C57BL/6鼠30只,每组10只,模型周期4周,对照组喂饲普通饮食;模型组喂含1%的胆固醇饮食;用药组喂含1%的胆固醇饮食和荜拔油不皂化物(30mg/kg体重)灌胃。结果:模型组9只出现胆囊结石,10只出现胆固醇结晶;荜拔油不皂化物组1只出现胆囊结石和结晶;对照组无结石和结晶形成。与模型组相比,荜拔油不皂化物组和对照组的肝氨肽酶N(APN)mRNA及胆囊胆汁胆固醇饱和指数(CSI)明显降低。结论:荜拔油不皂化物明显抑制高胆固醇饮食诱发的小鼠胆囊结石的形成,可能通过降低肝脏APN mRNA水平及胆囊胆汁胆固醇饱和指数发挥作用。