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.

Component Analysis and Free Radicals Scavenging Activity of Physalis alkekengi L. Polysaccharide

A crude polysaccharide was extracted from Physalis alkekengi L. fruit. HPLC was used for the component analysis of the polysaccharide. The results indicate that Physalis alkekengi L. polysaccharide（PAP） was composed of rhamnose, xylose, arabinose, galactose, and glucose. Free radicals scavenging activity of PAP was studied through 3 free radicals scavenging tests. PAP exhibited high scavenging effects on OH and DPPH radicals, and both the scavenging rates were about 80%. The scavenging rate of O2^- radical was about 22%.

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.

Pro-apoptotic and anti-proliferative effects of Physalis angulata leaf extract on retinoblastoma cells

AIM: To investigate the effect of Physalis angulata leaf extract on apoptotic and proliferation of retinoblastoma cells. Despite several previous studies evidencing the anticancer potential of Physalis angulata;however, certain study that proves its benefits in retinoblastoma cancer cells has been limited.METHODS: This study utilizes an in-vitro experimental study by applying Y79 human retinoblastoma cell line culture obtained from the American Type Culture Collection(ATCC;10801 University Boulevard Manassas, VA 20110, USA). The cell was divided into 4 groups. Group I was the control group without the administration of Physalis angulata leaf extract. Whereas, group II, II and IV are engaged with 25, 50, and 100 μg/mL of Physalis angulata leaf extract respectively. After a 24 h incubation, an examination with microtetrazolium(MTT) cell proliferation assay and Annexin V apoptosis detection was conducted. Statistical analysis was performed with the Tukey test.RESULTS: Physalis angulata leaf extract improved apoptosis and significantly reduced the number of living cells in retinoblastoma cells, along with the increase in the given dose. Based on the Tukey test, a significant difference was found in the treatment group at 50 μg/mL(P=0.025) and 100 μg/mL(P=0.001) in the measurement of apoptosis. Proliferation measurements also indicated a significant decrease in the number of living cells in the 50μg/m L treatment group(P=0.004), and in the 100 μg/mL treatment group(P=0.000). Meanwhile, a dose of 25 μg/mL indicated insignificant difference in the two measurements. Improved apoptosis and decreased number of living cells occured at a dose of 100 μg/mL. Decreased number of living cells(in the measurement of proliferation) was due to the inhibited proliferation or improved apoptosis.CONCLUSION: Physalis angulata leaf extract improve apoptosis in retinoblastoma cell culture, requiring further research to inhibit proliferation.

Assessment of antidiabetic activity and acute toxicity of leaf extracts from Physalis peruviana L.in guinea-pig

Objective:To verify the antidiabetic activity of leaf extracts from Physalis peruviana L.popularly used in the Eastern part of the Democratic Republic of the Congo and to point out the possible toxicity.Method:Aqueous decoctions prepared from dried leaves powder were administrated to guinea pigs at the dose range of 100 mg/kg to 3.2 g/kg of body weight.The hypoglycemic activity was evaluated by glucose tolerance test,loading animals with glucose 4 g/kg and measuring blood glucose concentrations at various times.The effect was compared to the control and glibenclamide as antidiabetic reference drug.Acute toxicity was evaluated by recording mortality rate,changes on blood biomarkers and damage caused to vital organs.Results:At a dose of 100 mg/kg,the aqueous extract induced a significant reduction of peak concentration at 30 min after glucose loading as compared with control or reference(P<0.05).At doses greater than 400 mg,some alterations on blood,kidney and liver markers were observed.Upper 800 mg/kg,mortality was observed with LD_(50)estimated at about 1280 mg/kg.At the autopsy,vital organs were in haemorrhage and swelling state.Conclusion:The crude aqueous extracts from the leaves of Physalis peruviana L.present hypoglycemic aclivily in animal model,but at high doses the plant may cause severe intoxication.

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.

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.

Effect of ethanolic extract of Physalis minima on lowering blood pressure in DOCA-salts hypertensive rat models

OBJECTIVE To explore the effect of ethanolic extract of Physalis minima on lowering blood pressure in the DOCA-salts hypertensive rats.METHODS To produce hypertension,male rats were induced DOCA at dose of 10mg·kg-1 ip.twice weekly and 0.9% NaCl in drinking water ad libitum for six weeks.The ethanolic extract of Physalis minima were given at dose of 500,1500 and 2500mg·kg-1,respectively for 4weeks.After completion of treatment schedule,systolic blood pressure was evaluated using animal blood pressure analyzer.The serum nitric oxide levels was measured using the ELISA methods.SOD activity and MDA level were determined spectrophotometrically.Endothelial dysfunction was determined by acetylcholine-induced vasorelaxation studies on isolated rat aortas.RESULTS The extract decreased systolic blood pressure significantly(P<0.001)in hypertensive rats.In addition,the extract also suppressed the production of MDA and restored SOD activity to normal level.There was a reduced relaxation response to acetylcholine in isolated aortic rings and serum NO level from hypertensive rats that were reversed by extract.CONCLUSION Physalis minima can lower blood pressure in hypertensive rats and its mechanism may be through the improvement of endothelial function due to the decrease of stress oxidative.

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.

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.

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.

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.

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.

Bionic Design and Experimental Validation of a Robotic Airship Inspired by the Physalia physalis

The robotic airship is one of the most unique and promising green aircraft,however,as a“lighter-than-air aircraft”and“thermal aircraft”,its long-endurance flight has great difficulties in decreasing drag and controlling buoyancy and pressure under thermal effects.In this work,we reported a robotic airship inspired by the Physalia physalis,imitating its morphology,physiological structure,and biological behaviors.The hull is designed by imitating the morphology of the Physalia physalis,and the gasbags including a helium balloon,two ballonets,and a thermoregulation gasbag are designed by imitating the physiological structure and biological behaviors of the pneumatophore,bladder,and gland of the Physalia physalis,respectively.Experimental results show that the bionic airship has an increase of about 40%in lift-to-drag and decreases the pressure in helium balloon by 47.5%under thermal effects,and has better aerodynamic performances and thermoregulation performances than conventional airships.

Chemical constituents from the fruit calyx of Physalis alkekengi var. francheti

In the present study, in order to investigate the chemical constituents of Physal＆ alkekengi L. var. franchetii （Mast.） Makino, the isolation of ingredients was performed by repeated chromatography on silica gel, Sephadex LH-20 and preparative HPLC. Their structures were identified based on 1D, 2D NMR, and mass spectral analysis, A total of 14 compounds were obtained, and their structures were identified as physalin P （1）, 4,7-didehydroneophysalin B （2）, physalin D （3）, 5α-hydroxy-25,27-dihydro-7- dehydro-7-deoxyneophysalin A （4）, 4,7-didehydrophysalin B （5）, ursolic acid （6）, wogonin （7）, blumenol A （8）, nobiletin （9）, liquiritigenin （10）, schizandrin （11）, 5-hydroxymethylfurfural （12）, 5-（hydroxymethy1）-2-（dimethoxymethyl）furan （13）, 1-O- [3-O-2-methyl-5-（2,3,4-trimethyl）pheny1-2,3-pentanedi]-β-D-xylopyranosyl-（1→6）-β-D-galactopyranoside （14）. Among them, compound 14 is a new compound. Compounds 7-11, 13 are isolated from Physalis alkekengi L. var.franchetii （Mast.） Makino for the first time.

Application of a chitosan coating as a carrier for natamycin to maintain the storage quality of ground cherry(Physalis pubescens L.)

Ground cherry(Physalis pubescens L.) is a kind of berry fruit favored by consumers in China; however, this fruit is particularly vulnerable to physiological senescence and pathogen attack during the traditional cold storage period. In order to maintain storage quality, a 1.5%(w/w) chitosan(CS) water solution containing 50 mg/L of natamycin(NA) was introduced. After all treatments were completed, the fruit was stored at 0°C and sampled every 10 d. At each sampling date, the following tests were performed: mold and yeast analyses; enzyme activity and content analyses which included superoxide dismutase(SOD), ascorbate peroxidase(APX), and malondialdehyde(MDA); and color analysis. In addition, a sensory evaluation was carried out for quality assessment at the end of the storage period. The results showed that the application of a chitosan coating combined with natamycin(CSNA) enhanced the activity of superoxide dismutase(SOD) and ascorbate peroxidase(APX), reduced the physiological rate of senescence, and inhibited pathogen attack. Thus, CSNA treatment can maintain ground cherries at an acceptable level of storage quality for 50 d.

Metabolomics Analysis of Health Functions of Physalis Pubescens L. using by Ultra-performance Liquid Chromatography/Electrospray Ionization Quadruple Time-of-Flight Mass Spectrometry

Herbal medicines may benefit from metabolomics studies, and applying metabolomics may provide answers about which herbal interventions may be effective for individuals, which metabolic processes are triggered, and the subsequent chemical pathways of activity. Physalis pubescens L(PPL) is an herbal fruit for one year living plant and has been developed into healthy function's food. However, the mechanisms of health functions are still unclear. To comprehensively and holistically assess its anti-fatigue and antioxidant effects, a novel integrative metabolomics approach was applied. In this study, we present metabolomics analysis applying ultra performance liquid chromatography coupled to quadrupole with time-of-flight mass spectrometry(UPLC-Q/TOF-MS) to determine metabolite alterations after oral administration PPL to rats. Fifteen metabolites in urine were identified as potential biomarkers. Pattern analysis of the UPLC-Q/TOF-MS data disclosed that PPL could relieve fatigue rats by ameliorating the disturbance in amino acids metabolism and energy metabolism, alleviating the oxidative stress from reactive oxygen species and the inflammatory damage, and recovering the destructed regulation. Based on these results, we demonstrated that PPL is a promising source of natural anti-fatigue and antioxidants material for use in functional foods and medicines.

Withaminimas A–F, six withanolides with potential anti-inflammatory activity from Physalis minima

Withaminimas A–F(1–6), six new withaphysalin-type withanolides were isolated from the aerial parts of Physalis minima L.. The structures of these compounds were elucidated through a variety of spectroscopic techniques including HR-MS, NMR, and ECD. Compound 1 belongs to rare 18-norwithanolides, and 2–3 were 13/14-secowithanolides. According to the traditional usage of P. minima, inhibitory effects on nitric oxide(NO) production in lipopolysaccaride-activated RAW264.7 macrophages were evaluated, and compounds 1–4 exhibited significant inhibitory effects with IC50 values among 3.91–18.46 μmol·L–1.

In vitro antileishmanial effects of Physalis angulata root extract on Leishmania infantum

Objective： In the present study, we evaluated the effects of the aqueous extract of Physalis angulata root （AEPa） on Leishmania infanturn proliferation, morphology, and the driving mechanism in leishmanicidal activity and modulatory action on macrophages. Methods： I., infantum promastigotes were treated with 50 and 100 μg/mL AEPa for 72 h and then antipro- mastigote assay was performed by counts in a Newbauer chamber, morphological changes were analyzed by transmission electron microscopy and the mechanism of the leishmanicidal activity was detected. In addition, macrophages were infected with L. infantum and were used to evaluate anti-amastigote activity of AEPa and effects of AEPa on cytokine secretion after 72-hour treatment. Results： Treatment with AEPa reduced the numbers ofL infantum promastigotes （50% inhibitory concen- tration （ICso） = 65.9μg/mL; selectivity index （SI）= 22.1） and amastigotes （ICso = 37.9 μg/mL; SI = 38.5） compared with the untreated control. Amphotericin B reduced 100% of the promastigote numbers after 72 h of treatment （IC50 = 0.2μg/mL）. AEPa induced several morphological changes and increased the production of reactive oxygen species and apoptotic death in promastigotes after treating for 72 h. AEPa （100 μg/mL） promoted tumor necrosis factor-α secretion in macrophages infected with L. infantum after 72 h of treatment, but did not induce an increase in this cytokine in noninfected macrophages. In addition, AEPa showed no cytotoxic effect on J774-A1 cells （50% cytotoxic concentration 〉1000μg/mL）. Conclusion： AEPa presented antileishmanial activity against the promastigotes and amastigotes of I.. infantum without macrophage cytotoxicity; these results show that natural products such as P. angulata have leishmanicidal potential and in the future may be an alternative treatment for leishmaniasis.

Modulative effect of Physalis alkekengi on both gut bacterial and fungal micro-ecosystem

Objective: Gut microbiome is an intricate micro-ecosystem mediating the human health and drug efficacy. Physalis alkekengi(PAL) is an edible and time-honored traditional Chinese medicine. Several pharmacological effects of PAL have been verified and gut bacteria are implied in its therapeutic actions.However, the detailed modulation of PAL on gut bacterial species and on gut fungi remains largely unknown. We, therefore, designed a preliminary experiment in normal mice to reveal the modulation effect of PAL on both gut bacteria and fungi, and explore the interaction between them.Methods: Herein, the aqueous extract of PAL was orally administrated to normal C57BL/6 mice for four weeks. The full-length 16S rRNA and ITS1/2 gene sequencing were explored to detect the taxa of gut bacteria and gut fungi after PAL treatment, respectively.Results: Oral administration of PAL notably enriched anti-infammatory bacterial species such as Duncaniella spp. and Kineothrix alysoides, whereas decreased pro-infammatory species such as Mucispirillum schaedleri. Simultaneously, PAL increased the abundance of gut fungi Aspergillus ochraceus,Cladosporium sp. and Alternaria sp., and decreased Penicillium janthinellum. Correlation network analysis identified two co-existing microbial groups(groups 1 and 2) that were negatively associated with each other. The group 1 comprised PAL-enriched bacteria and fungi, while group 2 was mainly normal chow-enriched bacteria and fungi. In group 1, Antrodia monomitica, Aspergillus clavatus, Mortierella kuhlmanii and Sarcinomyces sp. MA 4787 were positively correlated with Bifidobacterium globosum,Romboutsia ilealis and so on. In group 2, Chaetomium subspirilliferum, Septoria orchidearum and Cephaliophora tropica were positively related to Lactobacillus spp.Conclusion: Altogether, this preliminary study first demonstrated the modulation effect of PAL on both gut bacteria and gut fungi, which may shed light on the elucidation of PAL’s pharmacological mechanism.