Optimization of Polysaccharides Extraction from Physalis alkekengi L.Peel and Its Effect on the Expression of Inflammation-Related Proteins in SW620 Cells

Objective:To establish an optimized aqueous extraction process for polysaccharides from Physalis alkekengi L.peel and to preliminarily explore its in vitro anti-inflammatory activity against colorectal cancer SW620 cells.Methods:A single-factor test combined with orthogonal test analysis was used to evaluate the effects of the material-to-liquid ratio,extraction temperature,and extraction time on the yield of polysaccharides from Physalis alkekengi L.peel.The antioxidant activity of the polysaccharides was assessed by analyzing their free radical scavenging ability in vitro,and the anti-inflammatory effect was evaluated using SW620 cells.Results:The optimal extraction conditions were a material-to-liquid ratio of m(g):V(mL)=1:30,an extraction temperature of 100℃,and an extraction time of 40 minutes,with a predicted polysaccharide yield of 25.7%.The polysaccharides from Physalis peruviana peel effectively scavenged DPPH,superoxide anion,and hydroxyl radicals.After treatment with Physalis peruviana polysaccharides,the levels of IL-1β,IL-18,and TNF-αin the cell culture medium were significantly reduced,and the phosphorylation level of P65 protein in SW620 cells was decreased.Conclusion:This extraction method is stable and reliable,and the prepared Physalis alkekengi L.polysaccharides exhibit significant in vitro antioxidant and anti-inflammatory activities.This study provides a theoretical basis for developing drugs for the prevention and treatment of colorectal cancer.

A novel cytotoxic neophysalin from Physalis alkekengi var.francheti

A new neophysalin, named 5α-hydroxy-25,27-dihydro-4,7-didehydro-7-deoxyneophysalin A（1）, along with three other known neophysalins （2-4） were isolated from the calyxes of Physalis alkekengi L. var.francheti （Mast.） Makino. The structure of 1 was determined by means of 1D and 2D NMR, UV, IR and mass spectra. Compound 1 displayed potent cytotoxicities in vitro against PC- 3 and LNCaP cell lines.

Three new physalins from Physalis alkekengi var. franchetii

Three new physalin steroids,physalin Ⅲ(1),physalin IV(2),3-O-methylphysalin X(3),together with five known physalins(4-8)were isolated from the 80%EtOH extract of calyces of Physalis alkekengi var.franchetii.The structures of the new compounds were revealed through 1D and 2D NMR and mass spectroscopic studies.

Medicinal and Edible Plant——Physalis alkekengi L.and Its Cultivation Techniques

Botanical morphological characteristics of Physalis alkekengi L.is first introduced,and its medicinal and edible value is elaborated.Cultivation techniques of P.alkekengi L.are mainly introduced.The research aims to provide a certain basis for the better development and application of P.alkekengi L.

Study on Callus Induction and Plant Regeneration of Physalis alkekengi

[ Objective] This study aimed to investigate optimal conditions for callus induction and plant regeneration of Physalis alkekengi. [ Method ] P. alkek- eng/was employed as the experimental material and different concentrations of 6-BA and c^-NAA were added to MS medium to prepare the differentiation medium, to investigate the effect of different concentrations of plant hormones on callus differentiation of P. alkekengi. [ Result ] Under low or high concentrations of 6-BA, ratio of the total number of adventitious buds/explants and differentiation rate were reduced with the increasing concentration of ct-NAA ; when the concentrations of 6-BA and ct-NAA were respectively 1.0 and 0.2 mg/L, ratio of the total number of adventitious buds/explants and differentiation rate reached the maximum. [ Conclusion] When the concentrations of α-NAA were maintained unchanged, ratio of the total number of adventitious buds/explants reached the maximum under the moderate concentrations of 6-BA, while the differentiation rate showed irregular variations. In MS medium with supplement of 1.0 mg/L 6-BA and 0.2 mg/L α-NAA, ratio of the total number of adventitious buds/explants and differentiation rate reached the peak. This study provided reference for callus induction of P. alkekengi.

Polysaccharide from Fruits of Physalis Alkekengi L. Enhances Antitumor Efficacy by a DNA Vaccine

Physalis. alkekengi fruit has long been used in traditional Chinese medicine for tumor therapy. In the present study, using plasmids that encode ovalbumin (OVA) gene, we investigate the adjuvant activity of a polysaccharide fraction (PPSB) isolated from P.alkekengi fruit. Formulation by simple procedures of mixing of the OVA-encoding pCI-neo-sOVA plasmid with PPSB not only induced specific antibody responses, but also induced antigen-specific cytotoxic T lymphocyte (CTL) responses (Graph abstract). Furthermore, immunization using this vaccine prevented the growth of OVA-expressing B16-OVA tumor cell growth in the immunized mice. Thus, we provide evidence supporting the adjuvant activity of PPSB in DNA vaccine against tumor.

Intestinal anti-inflammatory activity of Ground Cherry(Physalis angulata L.)standardized CO2 phytopharmaceutical preparation

AIM To investigate the effects of Ground Cherry(Physalis angulata L.)standardized supercritical CO_2 extract in trinitrobenzenesulphonic acid(TNBS)model of rat intestinal inflammation.METHODS The animals were divided into groups that received vehicle or P.angulata extract(PACO_2)orally at the doses 25,50 and 100 mg/kg daily by 5 d before TNBS damage.Protective effects of PACO_2 were assessed by macroscopic analysis,biochemical determinations of the levels of myeloperoxidase(MPO),alkaline phosphatase(ALP),glutathione and cytokines(such as INF-γ,IL-1β,IL-6,IL-10 and TNF-α),gene expression evaluation(including Hsp70,heparanase,NF-κB,mitogenactivated protein kinases(Mapk)1,3,6 and 9,and the mucins genes Muc 1,2,3 and 4)and histopathological studies using optical,and electronic(transmission and scanning)microscopy.RESULTS PACO2 extract promoted a significant reduction in MPO and ALP activities,reducing oxidative stress and neutrophil infiltration.These effects were accompanied by significant reduction of colonic levels of IFN-γand IL-6 and down-regulation of heparanase,Hsp70,Mapk3,Mapk9,Muc1 and Muc2 genes expression when compared with TNBS-control animals.In addition,protective effects were also evidenced by reduced neutrophil infiltration,recovery of cell architecture and replacement of mucin by histopathological and ultrastructural analysis.CONCLUSION Physalis angulata supercritical CO2 extract is an intestinal anti-inflammatory product that modulates oxidative stress,immune response and expression of inflammatory mediators,with potentially utility for treating inflammatory bowel disease.

UPLC-MS/MS法测定苦蘵宿萼中physalin H和physalin D的含量

建立了同时测定苦蘵中2种酸浆苦味素类化合物含量的方法.应用超高效液相色谱(UPLC)与Qtrap质谱(MS)联用同时测定苦蘵中酸浆苦味素H(physalin H)和酸浆苦味素D(physalin D)2个成分的含量.采用Waters Acquity BEH C18(100 mm×2.1 mm,1.7μm)色谱柱,以水(0.1%甲酸)-乙腈(0.1%甲酸)为流动相,梯度洗脱,柱温35℃,流速0.3 mL·min^(-1),使用ESI+离子源,多反应监测(MRM)模式扫描,对physalin H和physalin D进行定量.结果表明physalin H和physalin D在5~10000 ng·mL^(-1)范围内具有良好的线性关系,相关系数(r)分别为0.9997和0.9995;平均加样回收率(n=5)分别为71.1%和71.0%,RSD值分别为1.36%和1.47%.该方法准确可靠,可用于苦蘵药材中physalin H和physalin D的含量测定,为有效控制苦蘵药材的质量奠定了基础.

Extraction and Crystal Structure of Physalin B

The title compound of physalin B（C28H32O9）, a main active physalin of Physalis angulata L, was isolated from the whole plant of Physalis angulata L, and characterized by X-ray diffraction analysis. It crystallizes in the monoclinic system, space group P21 with C28H32O9, a = 12.4996（2）, b = 14.35620（10）, c = 14.75190（10）, V = 2607.97（5） 3, Z = 4, Dc = 1.382 mg/cm3, Mr = 542.56, F（000） = 1152, and μ = 0.870 mm-1. The final R = 0.0389 and wR = 0.1037 for 47670 observed reflections（I 〉 2σ（I））. The rigid molecule consists of eight fused rings involving two lactones. There are two C28H32O9 molecules in an symmetric unit, and the title compound is stacked into a 3D layer structure through hydrogen bonds. In the 5~20 μmol/L range, physalin B can significantly inhibit the secretion of inflammatory cytokines TNF-α and IL-6 on RAW264.7 cells. The results suggest that physalin B has anti-inflammatory activity in vitro.

Methanolic extract of ceplukan leaf(Physalis minima L.)attenuates ventricular fibrosis through inhibition of TNF-α in ovariectomized rats

OBJECTIVE To investigate the effects of ceplukan leaf(Physalis minima L),which contain phytoestrogen physalin and withanolides,toward ventricular TNF-αlevel and fibrosis in ovariectomized rats.METHODS Wistar rats divided into six groups(K1:normal;K2:5 weeks ovariectomy(OVX);K3:9weeks ovariectomy(OVX),K4,K5,and K5:9weeks OVX+4 weeks ceplukan leaf's methanolic extract dose 500,1500,and 2500 mg·kg-1,respectively.TNF-αlevel measured with ELISA method.Fibrosis measured as blue color percentage in Masson′s Trichrome staining.RESULTS This study showed that prolonged hypoestrogen increase ventricular fibrosis(P<0.05).Ceplukan leaf treatment also resulted in a decreased ventricular fibrosis and TNF-αlevel in dose dependent manner compared with those of without treatment group(P<0.05).Furthermore,the TNF-αlevel normalized in rat treated with 2500mg·kg-1 Physalis minima L(P<0.05).Reduction of fibrosis positively correlated with TNF-αlevel(P<0.05,r= 0.873).CONCLUSION Methanolic extract of ceplukan leaf decrease ventricular fibrosis through inhibition of ventricular TNF-αin ovariectomized rats.Duration of hypoestrogen increase ventricular fibrosis.

A review of the ethnobotanical value,phytochemistry,and pharmacology of Physalis pubescens L.

Physalis pubescens L.(P.pubescens)was widely used for the treatment of inflammation-related diseases,such as sore throat,aphonia,phlegm,heat,and cough in folk medicine.The fruits of P.pubescens are commonly consumed as fruit in many areas of the world.In the past few decades,the phytochemistry and pharmacology of P.pubescens were extensively investigated.About 170 chemical constituents were purified from P.pubescens.The extract and chemical constituents of P.pubescens demonstrate diverse pharmacological effects,including anti-oxidation,anti-inflammation,anticancer,antimicrobial activity,immunomodulation,diuretic effect,hypoglycemic,and hypolipidemic in vitro and in vivo.Herein,we systematically summarized the ethnomedicinal uses,botanical characterization,distribution,phytochemistry,and pharmacology of P.pubescens,and establish the correlation between chemical constituents and pharmacological effects.

超声辅助溶剂提取锦灯笼宿萼总黄酮工艺的研究

锦灯笼宿萼富含多种生物活性成分,为了高效、充分提取锦灯笼宿萼总黄酮,以锦灯笼宿萼为原料,采用超声波辅助乙醇溶剂提取总黄酮,探讨超声波功率、提取温度、超声时间、乙醇体积分数及料液比对总黄酮得率的影响,并在单因素试验基础上采用L16(45)正交试验进一步优化提取工艺参数。结果表明,锦灯笼宿萼总黄酮提取的最佳工艺为:超声波功率180 W,提取温度60℃,用60%乙醇溶液按料液比1∶20(g/mL)浸泡锦灯笼宿萼粉末24 h后,超声提取50 min。在此条件下,锦灯笼宿萼总黄酮得率为2.982%。该工艺简单,制备成本低,便于工业化生产,有助于锦灯笼资源的综合利用和开发。

模糊数学结合响应面法优化酸浆多糖酸奶工艺

以酸浆多糖、纯牛奶为主要原料,加入发酵菌粉进行发酵,研制一款具有独特酸浆香气且营养丰富的酸奶。以感官评分为指标,在单因素试验基础上采用模糊数学评价法与响应面法相结合的方法,优选出酸奶生产工艺。结果表明,当酸浆多糖添加量为2.2%,木糖醇添加量为4.9%,菌粉添加量为0.15%,发酵时间为8 h时,参照此条件制作的酸浆多糖酸奶组织均匀,润滑细腻且香气浓郁,感官评分为87.17分,与预测值相近。

RP-HPLC测定酸浆药材中3种有效成分的含量

目的:采用 RP-HPLC 建立同时测定酸浆药材中酸浆苦味素 A、酸浆苦味素 D 及木犀草素含量的方法。方法:分析柱采用 Diamonsil C_(18)(200 mm×4.6 mm,5 μm)色谱柱,流动相为甲醇-0.2%醋酸(45:55,v/v);流速:1.0 mL·min^(-1);检测波长:220 nm。结果:酸浆苦味素 A、酸浆苦味素 D 及木犀草素浓度分别在0.15～1.20 mg·mL^(-1)(r=0.9995),0.10～0.81 mg·mL^(-1)(r=0.9996),0.12～0.96 mg·mL^(-1)(r=0.9997)范围内与峰面积呈良好的线性关系,酸浆苦味素 A 平均回收率分别为95.9%,97.1%,103%;RSD 分别为2.0%,0.7%,1.4%。酸浆苦味素 D 的平均回收率分别为97.3%,101%,97.1%;RSD 分别为1.0%,1.9%,1.3%。木犀草素的平均回收率分别为97.8%,97.8%,97.7%;RSD 分别为0.4%,2.9%,0.5%。结论:采用 RP-HPLC 同时测定酸浆药材中酸浆苦味素 A、酸浆苦味素 D 及木犀草素3种成分的含量,该法简单、可靠,可作为控制酸浆药材质量的参考标准。

锦灯笼果实泡腾片制备工艺研究

目的通过对处方组成和工艺进行优选制备锦灯笼果实泡腾片。方法以锦灯笼果实为原料,以甜菊糖苷、柠檬酸、碳酸氢钠、α-乳糖、聚乙二醇6000(PEG 6000)、聚乙烯吡咯烷酮(PVP)为辅料,采用酸碱混合制粒的方法制备锦灯笼果实泡腾片。并在以锦灯笼果实提取物添加量、甜菊糖苷添加量、崩解剂添加量等为考查因素进行单因素试验的基础上,利用响应面试验优化设计锦灯笼果实泡腾片的制备工艺。结果锦灯笼果实提取物的添加量为30%,甜菊糖苷添加量为0.15%,崩解剂添加量为50%(酸碱比为1∶1)作为最佳工艺条件,在此条件下制得的泡腾片片剂完整,表面呈浅咖色,崩解速度快,口感好,具有锦灯笼香气,苦甜适宜。结论该泡腾片的处方及制备工艺合理。

HPLC法测定锦灯笼果实中5种黄酮类成分含量

建立同时测定锦灯笼果实中5种黄酮类成分的高效液相色谱方法,为以后的锦灯笼质量评价研究做铺垫,并对5个不同产地的锦灯笼果实中黄酮含量进行测定。采用HPLC法,流动相甲醇-水,梯度洗脱;柱温30℃;流速1.0 m L/min;检测波长350 nm。结果表明5种黄酮类成分的线性关系良好,精密度、稳定性良好,符合测定要求。采用高效液相色谱法测定锦灯笼果实中5种黄酮类成分方法简便、准确,对5个不同产地样品测定结果稳定,重现性好。

HPLC法测定锦灯笼宿萼中槲皮素的含量

[目的]建立HPLC测定锦灯笼[Physalis alkekengi L.var.franchetii（Mast.）Makino]宿萼中槲皮素含量的方法。[方法]色谱柱为Diamonsil-C18（250.0 mm×4.6 mm,5μm）;流动相为甲醇-0.4%磷酸溶液（V∶V=55∶45）;流速为1.0 ml/min;进样量为20μl;检测波长为360 nm;柱温为50℃。[结果]槲皮素进样浓度在4.0~40.0μg/ml范围内,与峰面积呈良好的线性关系（r=0.999 6）;槲皮素平均回收率为98.17%,RSD值为0.46%。[结论]该方法简便、准确,可用于锦灯笼宿萼的质量控制。

NaCl胁迫对酸浆试管苗生理特性的影响

目的：研究NaCl对酸浆试管苗生长及生理特性的影响,以探讨其耐盐性。方法：在附加0.2%-1.0%NaCl的胁迫培养基上对试管苗进行培养,测定其脯氨酸、可溶性糖和可溶性蛋白、叶绿素、丙二醛（MDA）含量和质膜相对透性及超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和过氧化物酶（POD）的活性,并对其进行分析。结果：当NaCl为0.6%及以下时,随浓度的增加酸浆试管苗株高、茎粗、根长、生根数及叶绿素含量均逐渐降低,而当NaCl为0.8%及以上时,试管苗株高、茎粗和叶绿素含量急剧下降,根的发生被显著抑制;脯氨酸含量及SOD活性均随NaCl浓度的增加呈上升趋势,可溶性糖和可溶性蛋白及CAT和POD活性均随NaCl浓度的升高呈先上升后下降的趋势,但可溶性糖和可溶性蛋白含量均高于对照;MDA含量和质膜透性在0.6%NaCl及以下时积累缓慢,而NaCl为0.8%及以上时急剧增加。结论：酸浆试管苗具有适应一定浓度（≤0.6%NaCl）的盐渍生境能力,在其受到盐胁迫时可通过提高体内渗透调节物质含量和抗氧化酶活性来缓解盐害。

毛酸浆茎叶多糖的结构表征及体外活性

采用水提法从毛酸浆茎叶部分提取得到粗多糖,利用柱色谱分离纯化获得一个新的多糖(PPL-S1),对其化学结构、理化性质进行初步探究,并测定该多糖的体外抗氧化活性以及神经细胞保护活性。结果表明:多糖PPL-S1分子量为1 949.8 Da,单糖组成为鼠李糖、阿拉伯糖、甘露糖、半乳糖,其物质的量比为0.15∶0.21∶0.14∶0.50,具有多糖的特征吸收峰。体外自由基清除实验表明,PPL-S1对DPPH、ABTS自由基具有一定的清除效果,IC_(50)值分别为1.60 mg·mL^(-1)和1.43 mg·mL^(-1)。并且PPL-S1能显著改善H_(2)O_(2)诱导的氧化应激,提高神经细胞SH-SY5Y细胞存活率,对H_(2)O_(2)诱导的神经细胞损伤具有一定的保护活性。

灯笼果叶片、宿萼和果实化学成分分析及抗氧化活性评价

为全面评估分析酸浆属(Physalis)国内栽培最广的灯笼果(Physalis peruviana L.)化学成分及抗氧化活性,该研究采用紫外分光光度法测定了灯笼果叶片、宿萼和果实提取物总酚、总黄酮含量,应用超高效液相色谱质谱联用技术鉴定其化合物,并检测主要化合物含量,分别应用DPPH自由基、ABTS阳离子自由基、羟自由基清除和铁离子还原法测试提取物体外抗氧化活性。结果表明,灯笼果叶片提取物总酚(8.15 mg GAE/g DW)和总黄酮(30.43 mg RT/g DW)含量最高,宿萼提取物次之,果实提取物总含量最低。从3个部位中共鉴定4种黄酮、3种脂肪酸、1种醉茄内酯、1种二苯并呋喃类和1种羟基肉桂酸类化合物。叶片和宿萼提取物化合物种类丰富,其中叶片提取物紫云英苷(492.03μg/g DW)含量最高,宿萼提取物金丝桃苷(162.10μg/g DW)含量最高,果实提取物仅检测到阿魏酸-4′-O-葡萄糖苷化合物(137.37μg/g DW)。叶片提取物有良好的清除羟自由基能力(SC_(50)=0.61 mg/mL),宿萼提取物清除DPPH自由基(SC_(50)=0.19 mg/mL)、ABTS阳离子自由基能力(0.13 mmol TE/g)和铁离子还原能力(0.16 mmol Fe^(2+)/g)最优。研究结果丰富了灯笼果次级代谢产物数据,为充分利用灯笼果资源、开发天然抗氧化产品提供了科学依据。