Optimization of ISSR-PCR System and Conditions for Portulaca oleracea L.

With Portulaca oleracea L. as an experimental material, its total DNA was extracted by the improved CTAB method, the ISSR-PCR primers were screened, and the ISSR-PCR reaction system and reaction conditions for P. oleracea were Optimized. The results showed that there were 8 primers suitable for ISSR-PCR of P. oleracea. The optimal reaction system had a volume of 25 μl, including 2 x Taq Platinum PCR Master Mix 12.5 μl, primer 2 μl, ddH20 9.5 μl, and DNA template 1μl. The optimized ISSR-PCR of P. oleracea was started with pre-denaturation at 94 ℃ for 360 s, followed by 30 cycles of denaturation at 94 ℃ for 60 s, annealing at 54 ℃ for 60 s and extension at 72 ℃ for 90 s, and completed by extension at 72 ℃ for 300 s.

Chemical Constituents from Portulaca oleracea L.

Portulaca oleracea L. is distributed widely in China and some othercountries, which is included in the pharmacopoeias of China. In tradition, it is used in thetreatment of atheroma, and also used as antifungal and antiviral agents now. It also decreases bloodsugar, enhances immunity and adjusts blood lipids. The anticancer and antifungal effects ofPortulaca oleracea L. were reported in several patents. But no active component has been found. Inthis study, the chemical compositions from Portulaca oleracea L. were investigated to provideevidence for the activity of Portulaca oleracea L. .

Detection and quantification of cyclo-dopa amides in Portulaca oleracea L. by HPLC-DAD and HPLC-ESI-MS/MS

In the present study, we aimed to detect and quantify cyclo-dopa amides in Portulaca oleracea L. Together with four known ones （oleracein A-D）, we further identified eight new cyclo-dopa amides, named oleracein H-O, using an HPLC-DAD combined with an improved HPLC-ESI-MS/MS method. A standardized HPLC fingerprint of cyclo-dopa amides was generated for the first time through analyzing 11 batches ofP. oleracea. Simultaneously, the contents of two major amides, oleracein A （OA） and oleracein B （OB）, and the total amides （TID） were determined. Our results showed that the contents of OA and OB of P. oleracea ranged from 35.00 to 151.93 mg/kg and from 40.00 to 150.44 mg/kg, respectively. Moreover, TID ranged from 314.16 to 928.60 mg/kg （calculated by OA）, or from 475.83 to 1393.00 mg/kg （calculated by OB）. Taken together, our newly develooed method could be used for the aualitv assessment and oualitv control of this herb.

The Influence of Portulaca Oleracea L. Leaves Extracts on the Histoarchitecture of Culex Quinquefasciatus and Anopheles Stephensi Larvae

Background:Mosquito is a big threat to the human health.Mosquito-borne diseases cause millions of death to the human beings.Hence,a permanent solution is eagerly to be established to control its excessive growth in stagnant water.Portulaca oleracea Linn is a natural larvicidal agent,which contains active ingredients such as linolenic acid,linoleic acid(omega-3 fatty acids).These bioactive compounds may be responsible for its larvicidal properties on mosquito.Objective:The present study is focussed on identifying the bioactive compounds,such as linolenic acid,through GC-MS,and analyzing the larvicidal efficacy of Portulaca oleracea L.against Culex quinquefasciatus and Anopheles stephensi larvae.Methods:Preliminary phytochemical analysis,total protein content,total carbohydrate content,total phenol content,total flavonoid content and GC-MS analysis exhibited the presence of rich phytoconstituents in Portulaca oleracea.DPPH analysis was carried out to analyze the antioxidant potential of plant extract.Larvicidal activity and histological change were detected to evaluate the efficacy of Portulaca oleracea L.on the Culex quinquefasciatus and Anopheles stephensi larvae.Results:Aqueous and ethanol leaf extracts of Portulaca oleracea L.against both Culex quinquefasciatus and Anophe-les stephensi larvae,showed a very good larvicidal activity at 500μg/mL among the various concentrations.His-tological damages of mosquito larvae were observed when treated with the Portulaca oleracea L.extract,and provided further evidence for its larvicidal activity.Conclusion:This study concluded that the plant Portulaca oleracea L.contained many useful bioactive compounds,can be a strong larvicidal agent against both Culex quinquefasciatus and Anopheles stephensi larvae.The molecular mechanism for the larvicidal activity will be identified in future studies.

NaCl胁迫对马齿苋光合作用及叶绿素荧光特性的影响

以马齿苋为材料,采用温室盆栽法研究了14dNaCl胁迫处理对其幼苗生长、光合作用和叶绿素荧光特性的影响。结果显示:(1)马齿苋幼苗的鲜重和株高在25mmol.L-1 NaCl胁迫时与对照无显著差异,但其随着NaCl浓度的继续增加均显著降低,且其生物量受到的抑制早于株高。(2)NaCl胁迫下,马齿苋幼苗叶片净光合速率(Pn)降低,胞间二氧化碳浓度(Ci)增大,且两者的变化幅度随着NaCl浓度增加而增大。(3)NaCl胁迫下,马齿苋幼苗叶片的初始荧光(F0)、最大荧光(Fm)、可变荧光(Fv)、恒态荧光(Fs)、恒态荧光与初始荧光差值(△F0)、PSⅡ潜在光化学效率(Fv/F0)和PSⅡ最大光化学效率(Fv/Fm)均降低,叶片光化学荧光猝灭系数(qP)也在NaCl胁迫下降低,而非光化学荧光猝灭系数(NPQ)则上升;在0～50mmol.L-1 NaCl胁迫下,幼苗叶片各荧光参数下降幅度小于其他高浓度NaCl胁迫。研究表明,在NaCl胁迫条件下,马齿苋幼苗叶片的光合作用受光抑制伤害,但在低浓度NaCl下能够较多地将光能用于光化学反应,光抑制程度较低,保持了较高的净光合速率,明显减轻盐胁迫对植株生长的影响,表现出一定的耐盐性。

NaCl胁迫对马齿苋幼苗生长及体内离子分布的影响

以马齿苋幼苗为试验材料,研究了沙培条件下,0、100和200 mmol·L^(-1)NaCl处理对其生长及不同器官中Na^+、K^+、Ca^(2+)和Mg^(2+)分布的影响。结果表明:不同浓度NaCl处理下,马齿苋幼苗地上部干质量及单株叶片数均有所下降,但根系干质量和根冠比均有不同程度的增加。根、茎、叶中Na^+含量较对照均显著升高,且主要储存于根中;K^+含量较对照显著下降,茎中K^+含量降幅较小,而叶片中降幅较大;叶片中Ca^(2+)和Mg^(2+)含量显著高于茎和根。根、茎、叶中K^+/Na^+、Ca^(2+)/Na^+、Mg^(2+)/Na^+比值随NaCl处理浓度的增加而降低,主要原因是Na^+净增加;但叶片中K^+/Na^+、Ca^(2+)/Na^+、Mg^(2+)/Na^+比值显著高于茎和根系。随着Na Cl浓度的增加,马齿苋幼苗从根到茎的离子选择性运输能力先升高后降低,而茎到叶片的离子选择性能力总体呈升高趋势。综合分析表明,马齿苋可能通过将Na^+区域化到根系中,并增强植株向茎叶选择运输K^+、Ca^(2+)和Mg^(2+)的能力,从而缓解NaCl处理对马齿苋的伤害。

NaCl胁迫对马齿苋离子吸收及荧光特性的影响

以马齿苋为材料采用温室盆栽法,研究了NaCl胁迫处理对其生长发育、品质积累、离子分布和叶绿素荧光特性的影响。结果表明,马齿苋种子发芽率、幼苗的鲜质量和株高在25～50 mmol/L NaCl胁迫时比对照略有下降,但其随着NaCl浓度的继续增加均显著降低。0～75 mmol/L NaCl处理浓度可提高马齿苋幼苗维生素C含量。盐胁迫影响植物组织的离子分布,Na+浓度持续增加,K+、Ca2+和Mg2+浓度下降。NaCl胁迫下,马齿苋幼苗叶片的初始荧光(Fo)、最大荧光(Fm)、PSⅡ潜在光化学效率(Fv/Fo)和PSⅡ实际光化学效率(ФPSⅡ)均降低。但在0～50 mmol/LNaCl浓度下,各荧光参数与对照差异不显著。可见,NaCl胁迫下,马齿苋产生了光合作用的光抑制伤害。低浓度下,植株能够较多地将光能用于光化学反应,光抑制程度较低,保持了较高的净光合速率,最终明显减轻盐胁迫对植株生长的影响。

RP-HPLC测定马齿苋黄酮提取物中染料木苷的含量

目的检测马齿苋黄酮提取液中染料木苷的含量。方法用反相高效液相色谱（RP-HPLC）检测马齿苋黄酮部位中染料木苷的含量。色谱柱为SunfireC18column（250.0mm×4.0mm，5.0μm），流动相为甲醇：冰醋酸：水＝45：1：55，流速为0.8ml/min，柱温25℃，柱压2144psi，检测波长为259nm。结果马齿苋黄酮部位中染料木苷与标准品中染料木苷保留时间一致，染料木苷进样量在0.0625～2.0000ng范围内线性关系良好（r＝0.9999），平均回收率98.30％，RSD＝0.57％（n＝5）。结论RP-HPLC测定方法简便，结果可靠，含生药6kg/L的马齿苋黄酮提取液中染料木苷含量为1.66mg/L。

马齿苋水提物中生物碱oleraisoindole和7’-乙氧基-反式-阿魏酰基酪胺及其抗氧化活性研究

目的:研究马齿苋(Portulaca oleracea L.)水提取物中生物碱类化学成分分离及其抗氧化活性。方法:依次采用水提取、乙酸乙酯萃取、硅胶柱色谱、ODS柱色谱进行提取、分离及纯化,采用超高效液相色谱进行制备和核磁共振技术进行结构鉴定。然后,采用DPPH自由基清除的方法评估生物碱类化合物抗氧化活性。结果:通过1H-NMR、13C-NMR与有关文献对照,证明两种化合物分别为oleraisoindole和7’-乙氧基-反式-阿魏酰基酪胺(ETFT)。结论:马齿苋中分离得到两种生物碱oleraisoindole和ETFT,并首次开展了两种生物碱的抗氧化活性研究,结果表明两种生物碱具有显著的抗氧化活性。

响应面法优化马齿苋总多酚提取工艺及其抗氧化活性分析

以马齿苋为原料,采用超声辅助提取马齿苋总多酚。以总多酚提取量为指标,在单因素试验的基础上通过响应面法优化提取工艺,并分析马齿苋总多酚的抗氧化活性。结果表明:最佳提取工艺为液料比30:1(mL/g)提取温度45℃、乙醇体积分数60%、提取时间40min,在此条件下马齿苋总多酚提取量为6.89mg/g。抗氧化试验表明,马齿苋总多酚具有较好的抗氧化活性。

Isolation and Characterization of Endophytic Fungi from Purslane and the Effects of Isolates on the Growth of the Host

Purslane, a common weed, has been used as food or folk medicine in many countries. The growth, medicinal components and nutrient contents of the plant are closely associated with endophytes, especially endophytic fungi. In this study, the endophytic fungi associated with purslane were isolated, and the effects of the isolates on the host were investigated to lay a foundation for further research and development of purslane resources. The results showed that a total of eight endophytic fungi were isolated from purslane (collected from Hohhot, Inner Mongolia, China), and they belonged to the genera Penicillium (isolates K, N, P, M and I), Chaetomium (isolate J), Fusarium (isolate H) and Petriella (isolate O). Moreover, the growth of purslane was significantly influenced by its endophytic fungi. Isolate M can significantly decrease the germination rate, while J can significantly increase the germination rate of purslane. In addition, H, J and M can significantly increase the bud length of purslane, and the fermentation broth of P has a negative influence on the bud length of purslane. M and I can significantly increase the height, fresh weight and chlorophyll content of purslane due, in part, to the lower pH of the fermentation broth of I and M.

马齿苋活性成分及在化妆品中的功效

马齿苋(Portulaca oleracea L.)是一种常见的中草药植物,含有黄酮类化合物、生物碱类化合物、有机酸类化合物等丰富的活性成分,具有抗菌、抗炎、抗氧化、抗衰老、抗过敏等药理活性,且已被应用于化妆品中。对马齿苋活性成分及在化妆品中的功效等方面的研究进展进行了综述,以期为今后马齿苋的深入研究和开发应用提供理论支持。

Portulaca oleracea alleviates CCl4-induced acute liver injury by regulating hepatic S100A8 and S100A9

Objective: Acute liver injury(ALF) is a potential factor of many serious hepatopathies. Carbon tetrachloride(CCl4) is a possible environmental toxicant that can induce ALF. Portulaca oleracea(PO) is one of the most popular edible herbs and has several biological activities such as antioxidant, antimicrobial, antiinflammatory effects. We explored the significance of PO in regulating inflammatory function in animal models and cultured hepatocytes during liver damage caused by CCl4.Methods: The effect of PO on ALF was evaluated by CCl4-induced mice models in vivo. Hepatic levels of transaminase activities and inflammatory factors were examined. The gene and protein expression of S100A8 and S100A9 were measured by RT-PCR and Western blot analysis. Meanwhile, the efficacy of PO was certified by HepG2 cells in vitro. The transaminase activities, inflammatory factors, and the protein expression of S100A8 and S100A9 were also detected.Results: Animal tests showed that pretreatment with PO reduced the liver pathological tissue damage and the serum levels of ALT, AST, ALT and LDH, as well as reducing the pro-inflammatory cytokines(IL-1β, IL-6, TNF-a) secretion in CCl4-induced liver injury mice. Simultaneously, Hep G2 cells pretreated with PO exhibited a significant decrease in the activities of ALT and AST. Moreover, PO resulted in a significant downregulation of the pro-inflammatory markers S100A8, S100A9 gene and protein expression on CCl4induced acute liver injury was demonstrated entirely in vivo and vitro experiments.Conclusion: PO may down-regulate S100A8 and S100A9 and inhibit pro-inflammatory cytokines’ release,indicating a potential clinical effect for controlling the disease.

马齿苋中抗炎活性物质的提取、分离及结构鉴定

以活性物质示踪为导向,建立脂多糖诱导的RAW264.7巨噬细胞炎症模型对马齿苋中的抗炎物质进行跟踪,采用柱层析提取法、硅胶柱色谱分离法、制备液相色谱法及气相色谱-质谱联用技术对抗炎物质进行提取分离和结构鉴定。结果表明,石油醚-乙醇、无水乙醇和纯水溶剂依次对马齿苋样品进行提取,三种粗提物将细胞中一氧化氮(Nitric Oxide,NO)的分泌量分别减少至33.13、25.83和20.53μmol/L,其中石油醚相粗提物的抑制效果最强(P<0.05)。对石油醚相进一步分离得到四个组分,Fr.1、Fr.2和Fr.3组分具有较强的抗炎效果,但Fr.1和Fr.2组分含有潜在的毒性成分,选择Fr.3组分继续分离。Fr.3组分经硅胶柱分离得到三个组分,Fr.3.1组分表现出最强的抑制NO的分泌量效果(11.80μmol/L)。经制备液相色谱进一步纯化及气质分析,确定Fr.3.1组分的主要成分为硬脂酸(47.09%)、邻苯二甲酸二(2-乙基己)酯(13.21%)和其他成分。该研究建立了一种从马齿苋中分离纯化出抗炎物质方法,为马齿苋的开发利用提供理论参考。

Extracts of Portulaca oleracea promote wound healing by enhancing angiology regeneration and inhibiting iron accumulation in mice

Objective:To investigate the role of Portulaca oleracea(POL)in promoting revascularization and reepithelization as well as inhibiting iron aggregation and inflammation of deep tissue pressure injury(DTPI).Methods:The hydroalcoholic extract of POL(P)and aqueous phase fraction of POL(PD)were prepared based on maceration and liquid–liquid extraction.The number of new blood vessels and VEGF-A expression level were assessed using H&E stain and Western blot on injured muscle to examine the role of POL different extracts in vascularization.The iron distribution and total elemental iron of injured muscle were detected using laser ablation inductively coupled plasma mass spectrometry(ICP-MS)and Perls’staining to determine whether POL extracts can inhibit the iron accumulation.Besides,the ability of POL extracts to promote wound healing by combining re-epithelization time,inflammation degree and collagen deposition area were comprehensively evaluated.Results:In vitro,we observed a significant increase in HUVEC cell viability,migration rate and the number of the tube after P and PD treatment(P<0.05).In vivo,administration of P and PD impacted vascularization and iron accumulation on injured tissue,evident from more new blood vessels,higher expression of VEGF-A and decreased muscle iron concentration of treatment groups compared with no-treatment groups(P<0.05).Besides,shorter re-epithelization time,reduced inflammatory infiltration and distinct collagen deposition were associated with administration of P and PD(P<0.05).Conclusion:POL extract administration groups have high-quality wound healing,which is associated with increased new blood vessels,collagen deposition and re-epithelization,along with decreased iron accumulation and inflammatory infiltration.Our results suggest that that POL extract is beneficial to repair injured muscle after ischemia–reperfusion,highlighting the potential of POL in the DTPI treatment.

低糖马齿苋紫薯酱加工工艺优化及其产品品质评价

为了满足消费者对口味多样化、健康饮食的需求,开发一种低糖、零防腐剂的新型复合果酱的科学配方和先进制造方法。以富含人体健康功能因子的马齿苋和紫薯为主要原料,通过单因素实验和正交试验优化了马齿苋紫薯酱的配方,并对最终产品及其贮藏期间的营养物质、理化和卫生指标进行了测定。结果表明,低糖马齿苋紫薯酱的最佳配方为:马齿苋茎叶汁紫薯块根浆质量比为5:5,柠檬酸添加量为0.3%,山梨糖醇添加量为25%,增稠剂添加量为0.4%。按此配方制得的马齿苋紫薯酱色泽呈紫红色,酱体均匀,口感细腻,酸甜适中,具有马齿苋和紫薯特有的香味,感官评分为90.5±0.4分。最终产品的可溶性固形物为42.32%±1.43%,总糖为13.41±0.52 g/100 g,pH3.61±0.18,粗蛋白为0.83±0.08 g/100 g,VC为3.77±0.11 mg/100 g,黄酮为4.18±0.10 mg/g,多酚为2.05±0.13 mg/g,花青素为37.27±0.84 mg/100 g,以上理化指标及卫生指标均达到果酱类的国家标准。马齿苋紫薯酱对DPPH自由基的IC50值为7.071 mg/mL,表明该酱具有一定的抗氧化活性。马齿苋紫薯酱分别在25℃和37℃条件下贮藏1个月后,感官评分及各营养成分均有一定程度下降,菌落总数有所增加,但仍符合果酱国家标准。本项研究结果可为马齿苋和紫薯的精深加工及马齿苋紫薯酱的标准化生产提供参考依据。

基于网络药理学和分子对接探讨马齿苋改善肝损伤的分子作用机制

目的基于网络药理学及分子对接技术探讨马齿苋改善肝损伤的有效活性成分及分子机制。方法通过Traditional Chinese Medicine Systems Pharmacology(TCMSP)数据库检索马齿苋的活性化合物及相关靶点;GeneCards、On⁃line Mendelian Inheritance in Man(OMIM)数据库收集肝损伤的相关靶点;通过Venny 2.1.0网站筛选马齿苋与肝损伤的共表达靶点。运用Cytoscape 3.9.1软件和Search Tool for the Retrieval of Interacting Genes/Proteins数据库构建马齿苋活性成分与肝损伤共表达靶点的“药物-主要活性成分-共表达靶点-疾病”网络以及蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络图。通过the Database for Annotation,Visu⁃alization and Integrated Discovery数据库对共表达靶点进行基因本体论(gene ontology,GO)富集和京都基因与基因组百科全书(Kyoto encycloppedia genes and genomes,KEGG)通路注释分析,阐述马齿苋改善肝损伤的作用通路及分子作用机制。通过CB-Dock在线分子对接网站验证马齿苋有效活性成分与关键靶点之间的关系。结果从TCMSP数据库中筛选出10种马齿苋活性化合物及其对应的78个靶点,从GeneCards和OMIM数据库中筛选出1489个肝损伤的相关靶点,通过Venny 2.1.0网站找到54个马齿苋活性成分与肝损伤的共表达靶点。通过Cytoscape 3.9.1软件获得“药物-主要活性成分-共表达靶点-疾病”网络并找出马齿苋改善肝损伤的主要活性成分,在PPI网络图中得到关键靶点为TNF、IL6、IL1B、MMP9、CASP3、IL10、PPARG。GO富集分析结果显示马齿苋主要对转录、凋亡等生物过程起调节作用,通过KEGG通路分析筛选出与马齿苋改善肝损伤关系密切的通路为Non-alcoholic fatty liver disease通路,并且其重要的靶点有TGFB1,INSR,TNF,TNFRSF1A,IL1A,CASP7,IL6,MAPK8,CASP8,IL1B,CASP3,PPARG和PPARA。分子对接结果表明,马齿苋中主要活性成分β-胡萝卜素、槲皮素与核心靶点TNF、CASP3、PPARG稳定结合。结论马齿苋中的β-胡萝卜素、槲皮素通过与TNF、CASP3、PPARG结合调节Non-alcoholic fatty liver disease等信号通路发挥改善肝损伤的作用。

马齿苋中吲哚类生物碱的生物活性研究

目的:研究马齿苋中分离所得吲哚类生物碱的抗胆碱酯酶活性及抗氧化活性。方法:采用改良的Ellman法测定各化合物的抗胆碱酯酶活性,通过测量吸光度值计算化合物的胆碱酯酶抑制率,并计算IC_(50)值;采用DPPH自由基清除法测定各化合物的抗氧化活性,通过测量吸光度值计算化合物的DPPH清除率,并计算IC_(50)值。结果:马齿苋中oleraindole A-oleraindole G均有抗胆碱酯酶活性,其中,oleraindole B抗胆碱酯酶作用的IC_(50)值为40.22μM,与阳性药毒扁豆碱(IC_(50)=37.92μM)的活性相当;oleraindole A、B、C、D和G均具有抗氧化活性,其中,oleraindole C和oleraindole A抗氧化作用的IC_(50)值分别为7.49μM和9.68μM,远远优于阳性药BHA的活性(IC_(50)=37.81μM)。结论:oleraindole B抗胆碱酯酶活性较强,oleraindole C和oleraindole A抗氧化活性较优,可为马齿苋生物碱的深入研究提供科学依据。

Research and application of Portulaca oleracea in pharmaceutical area

Portulaca oleracea, a plant species used as medicine and food, is widely spread in most areas of China. It is also a popular food in some Europe and Mediterranean countries. Purslane contains a variety of chemical constituents with pharmacological activities, such as antitumor, hypoglycemic, antioxidant, antibacterial,anti-inflammatory activities. It is also full of plenty of nutrients serving as a diet food or healthy food.Here we reviewed purslane in the following aspects: botanical resources, ethnopharmacological function,chemical constituents, pharmacology and pharmacokinetics, safety evaluation and toxicity, and clinical applications.