Crude Polysaccharide Extract Research Articles

Chemical characteristics and bioactivity potential of polysaccharide extracts and purified fractions from Arctic brown macroalgae.

Macroalgae growing in the polar regions are exposed to extreme environment conditions and may induce differences in the structural and bioactive properties of their polysaccharides. Six brown macroalgae viz. kelp species - Saccharina latissima, Laminaria digitata, and Alaria esculenta; rockweed Fucus distichus; and filamentous macroalgae - Chorda filum and Chordaria flageliformis, from the Arctic were investigated for polysaccharides and their bioactivity. Among them, C. filum and C. flageliformis are least studied. Crude polysaccharide extracts (CPEs), extracted in water using ultrasonication, were found be dominated by fucoidan and alginate based on their monosaccharide composition analyzed using HPAEC-PAD. The CPEs showed up to 66% radical scavenging activity in DPPH, ABTS and H2O2 antioxidant assays, while 93% inhibition activity was observed in α-glucosidase anti-glycemic assay. The CPEs lacked pigments but contained minor amount of polyphenols. Purified polysaccharide fractions (PPFs) were obtained from the CPEs by ion-exchange chromatography. The monosaccharide composition and colorimetric analysis indicated enhanced purity of polysaccharides and absence of polyphenols in the PPFs. However, the antioxidant and antiglycemic activities of PPFs were lower than CPEs. We attributed the higher bioactivities in the crude extracts to synergistic interactions that may stabilize key active constituents.

Utilization of electron beam irradiation pretreatment for the extraction of pectic polysaccharides from Diaphragma juglandis fructus: Structural, physicochemical, and functional properties

The effects of electron beam irradiation (EBI) pretreatment on the alkaline extraction of pectic polysaccharides from Diaphragma juglandis fructus (DJF) are highly dependent on the irradiation dosage. Comprehensive characterizations encompassing physicochemical, structural, and functional properties were conducted on crude pectic polysaccharide extract from DJF subjected to various EBI doses. EBI pretreatment significantly increased the yields of crude pectic polysaccharides extract (increasing by 41.89 %), also facilitating the extraction of uronic acid, RG-I structure, and protein content, despite causing a decrease in total sugar content. EBI pretreatment induced the degradation of pectin, resulting in decreased molecular weight, particle size, crystallinity, viscosity, thermal stability, and water holding capacity, while enhancing solubility and oil holding capacity. Variations in physicochemical and structural properties induced by different EBI doses influenced the functional activities of DJF pectic polysaccharides. Low-dose EBI (at 5 kGy) pretreatment markedly improved the emulsifying activity/stability (increasing by 20.82/74.10 %) and ABTS/DPPH radical scavenging activity (increasing by 27.91/12.40 %), whereas high-dose EBI pretreatment (50 kGy) greatly enhanced foaming capacity/stability (increasing by 259.99/175.56 %). These findings provide a novel regulatory strategy for the functional activity of pectic polysaccharides.

Clinacanthus nutans crude polysaccharide extract as a green platform for microwave-assisted synthesis of silver nanoparticles: Optimization, characterization, and evaluation of bioactivities

Clinacanthus nutans (C. nutans) is a plant in tropical Asia with proven biological activities. The optimized extraction method of C. nutans crude polysaccharide (CNP) uses water in the presence of an ultrasound-assisted mechanical method (UL_CNP). However, the use of UL_CNP for the synthesis and optimization of silver nanoparticles (AgNP), particularly their anticancer and photocatalytic properties, remains unexplored. Hence, this research aimed to employ a green method using UL_CNP and silver nitrate to produce AgNP (UL_AgNP) with a small size and assess its potential toxicity, anticancer, and photocatalytic activities. The synthesis condition was optimized using the Box–Behnken design method. The synthesized UL_AgNP showed the surface plasmon resonance peak at 458 nm. The optimized synthesis condition produced spherically shaped UL_AgNP with a size of 5.21 ± 1.92 nm and a zeta potential of −26.33 ± 0.93 mV. An X-ray diffraction analysis exhibited intense Bragg's reflection peaks at (111), (200), (220), and (311), having a face-centered cubic structure of AgNP. Attenuated total reflectance-Fourier-transform infrared spectroscopy and energy-dispersive X-ray spectroscopy further confirmed the presence of silver in the synthesized UL_AgNP. The brine shrimp lethality test of UL_AgNP reported a lethal concentration 50 value of <7.8 μg/mL after 24 h. The UL_AgNP exhibited antiproliferative activity against MCF-7 cells with a half-maximal inhibitory concentration value of 4.96 ± 0.31 μg/mL by inducing S-phase cell cycle arrest, apoptotic effect, and reduction of cell migration. Furthermore, UL_AgNP proved its efficient photocatalytic activity against methylene blue dye (50.22 % ± 0.06 %, after 10 min at a concentration of 50 μg/mL). Therefore, the UL_AgNP exhibited promising antiproliferative activity against MCF-7 cells, highlighting their potential as a therapeutic agent. Further investigations are needed to elucidate the precise mechanism of their action.

Silver Nanoparticles with Modified Synthesis Use Sodium Borohydride Reducing Agent and Okra (Abelmoschus Esculentus L.) Raw Polysaccharide Extract as an Anti-Colon Cancer

Nanomedicine using nanoparticles has become a new trend in the treatment of colon cancer. Nanosilver is known to have antibacterial, antifungal, antidiabetic, and anticancer properties. The most common and easy way to make nanosilver is using the reducing agent sodium borohydride, but this inorganic compound element has high toxicity to the body. Polysaccharides have long been known as anticancer agents with low toxicity and few side effects. Okra (Abelmoschus esculentus L.), a flowering plant from the Malvaceae family found in tropical and subtropical areas, and the crude polysaccharide extract from the pods has the highest polysaccharide content. This research aims to determine the effect of raw okra polysaccharide extract (ORPE) as a reducing agent for making nanosilver on its ability as an anti-colon cancer cell line HCT 116. Experiments were carried out by modifying and optimizing the manufacture of AgNPs by reducing NaBH4 with ORPE, 16 concentrations each with repetition 3 times. Characteristic tests were carried out using UV-Vis Spectrophotometry, Particle Size Analyzer, Scanning Electron Microscopy, Transmission Electron Microscopy, cell viability testing with MTT assay, and evaluation of potential apoptosis and necrosis using Annexin V-PI flow cytometry. PSA test for average particle size. Two groups 1 (AgNP-NaBH4) and 2 (AgNP-ORPE) each had repeated measurements 3 times in 16 concentrations. The mean PSA test and zeta potential value for group 1 = 232.5 ± 25.47 nm / –42.23 ± 1.45 mV and 2 = 779.66 ± 112.45 nm / –23.15 ± 3.65 mV. TEM showed that the size of Group 1 was 50.85 nm (ꭓ = 113.14 nm) and Group 2 was 121.43 nm (ꭓ = 248.52 nm). SEM showed that the morphology of both groups was round in shape (group 2 with slight agglomeration). The absorbance spectrum is formed at a wavelength of 389 nm (group 1) and 281.5 nm (group 2). The IC50 value obtained by group 1 = 76.68 mmol/L with 60.3 % apoptotic cells, 3.74 % necrosis and group 2 = 92.58 mmol/L with 81.4 % apoptotic cells and 4.95 % necrosis. ORPE as a nanosilver-reducing agent has been proven to have the potential to induce cell death and cause changes in mitochondrial membrane permeability in the intrinsic pathway of apoptosis.

Inhibition of dengue virus infection in vitro by fucoidan and polysaccharide extract from marine alga Sargassum spp.

Dengue virus (DENV) infection poses a global health threat, leading to severe conditions with the potential for critical outcomes. Currently, there are no specific drugs available whereas the vaccine does not offer comprehensive protection across all DENV serotypes. Therefore, the development of potential antiviral agents is necessary to reduce the severity risk and interrupt the transmission circuit. The search for effective antiviral agents against DENV has predominantly focused on natural resources, particularly those demonstrating diverse biological activities and high safety profiles. Cyanobacteria and algae including Leptolyngbya sp., Spirulina sp., Chlorella sp., and Sargassum spp., which are prevalent species in Thailand, have been reported for their diverse biological activities and high safety profiles. However, their anti-DENV activity has not been documented. In this study, the screening assay was performed to compare the antiviral activity against DENV of crude polysaccharide and ethanolic extracts derived from 4 species of cyanobacteria and algae in Vero cells. Polysaccharide extracts from Sargassum spp. were the most effective in inhibiting DENV-2 infection under co-infection conditions, where the virus was exposed to the extract at the time of infection. Treatment of the extract significantly reduced the ability of DENV to bind to the host cells to 47.87 ± 3.88 % while treatment upon virus binding step had no antiviral effect suggesting the underlaying mechanism of the extract on interfering virus binding step. Fucoidan, a key bioactive substance in Sargassum polysaccharide, showed to reduce DENV-2 infection to 26.59 ± 5.01 %, 20.46 ± 6.58 % under the co-infection condition in Vero and A549 cells, respectively. In accompanied with Sargassum polysaccharide, fucoidan disturbed the virus binding to the host cells. These findings warrant further development and exploration of the Sargassum-derived polysaccharide, fucoidan, as a promising candidate for combating DENV infections.

Crude Polysaccharide Extract from the Microalga Porphyridium cruentum Improved Nonspecific Immune Responses and Resistance in Penaeus vannamei Exposed to Vibrio alginolyticus

Sulfated polysaccharides (SPs), especially those with immunostimulant effects on shrimp and fish, are bioactive compounds with potential use in aquaculture. In this study, Pacific white shrimp (Penaeus vannamei) were fed diets containing 0%, 0.5%, 1.0%, 1.5%, or 2.0% crude extracts from the microalga Porphyridium cruentum for 30 days. After dietary supplementation, the shrimp were exposed to Vibrio alginolyticus, allowing the assessment of phenoloxidase activity (PO) and total hemocyte count (THC) pre- and post-challenge, as well as shrimp survival rates. Shrimp that received the 1.0% treatment had greater survival (90%) at 48 h post-challenge, while the 0% and 2.0% treatments had survival rates close to 63% in the same period. Furthermore, at 48 h post-challenge, THC was significantly greater (p &lt; 0.05) in the shrimp that received crude extract supplementation compared to that in shrimp of the control group (0%). PO showed a proportional increase according to crude extract inclusion levels. Such dose-dependency in response to PO increases became significantly greater (p &lt; 0.05) in the 2.0% treatment group compared to that in the 0% treatment group in the pre-challenge period and that in the 0% and 0.5% treatment groups in the post-challenge period. These results showed that crude extracts from P. cruentum could be considered immunostimulants by increasing the resistance of P. vannamei exposed to V. alginolyticus, potentially representing a major breakthrough for marine shrimp farming.

Screening of Antioxidative and Antiproliferative Activities of Crude Polysaccharides Extracted from Six Different Plants

Plant polysaccharides have gained interest in medical research for their ability to suppress various diseases, including cancer. However, information on some plant polysaccharides is yet to be uncovered. In this study, we screened crude polysaccharides extracted from six different plants for their antioxidative and antiproliferative activities. Crude polysaccharides were isolated from different parts of some plants using the established extraction protocol. The crude polysaccharides were evaluated for their chemical composition (protein, total sugar, and phenolics), free radical-scavenging activities, and antiproliferative activities against breast cancer MCF-7 cells as well as non-cancerous cells, i.e., human fibroblast MRC-5 cells and Cercopithecus aethiops kidney Vero cells, via an MTT assay and CM20 Incubation Monitoring System (Olympus) for MCF-7. The investigated crude polysaccharides showed significant variations in their chemical constituents and antioxidative properties. Only Moringa seed crude polysaccharide extracts showed significant antiproliferative activities at various concentrations, with an IC50 value of 0.061 mg/mL, which was about 2.6 folds higher on MRC-5 and Vero cell lines. The antiproliferative activities toward cancer cell lines and lack of significant toxicity in the case of normal cells indicate that this extract may be promising as a valuable source for novel cancer therapy.

Physicochemical Properties and Biological Activities of Quinoa Polysaccharides.

Quinoa, known as the "golden grain" for its high nutritional value, has polysaccharides as one of its sources of important nutrients. However, the biological functions of quinoa polysaccharides remain understudied. In this study, two crude polysaccharide extracts of quinoa (Q-40 and Q-60) were obtained through sequential precipitation with 40% and 60% ethanol, with purities of 58.29% (HPLC) and 62.15% (HPLC) and a protein content of 8.27% and 9.60%, respectively. Monosaccharide analysis revealed that Q-40 contained glucose (Glc), galacturonic acid (GalA), and arabinose (Ara) in a molar ratio of 0.967:0.027:0.006. Q-60 was composed of xylose (xyl), arabinose (Ara), galactose, and galacturonic acid (GalA) with a molar ratio of 0.889:0.036:0.034:0.020. The average molecular weight of Q-40 ranged from 47,484 to 626,488 Da, while Q-60 showed a range of 10,025 to 47,990 Da. Rheological experiments showed that Q-40 exhibited higher viscosity, while Q-60 demonstrated more elastic properties. Remarkably, Q-60 showed potent antioxidant abilities, with scavenging rates of 98.49% for DPPH and 57.5% for ABTS. Antibacterial experiments using the microdilution method revealed that Q-40 inhibited the growth of methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli), while Q-60 specifically inhibited MRSA. At lower concentrations, both polysaccharides inhibited MDA (MD Anderson Cancer Center) cell proliferation, but at higher concentrations, they promoted proliferation. Similar proliferation-promoting effects were observed in HepG2 cells. The research provides important information in the application of quinoa in the food and functional food industries.

Effect of Lactiplantibacillus plantarum fermentation on the physicochemical, antioxidant activity and immunomodulatory ability of polysaccharides from Lvjian okra

Okra polysaccharides exhibits a range of biological activities. To date, its processing using microbial fermentation has not been explored. This study investigated the fermentation of okra juice with various lactic acid bacteria, followed by the extraction and characterization of crude polysaccharides (termed OPS-F), in contrast to their non-fermented counterpart (OPS). Changes in physicochemical properties, antioxidant activity and immunomodulatory ability were noted. The results demonstrated that OPS-F had a 7.42–12.53 % increase in total polysaccharides content compared to OPS. However, high-performance size-exclusion chromatography indicated a reduction in the molecular weight of OPS-F (7.9–9.5 × 105 Da) relative to OPS (1.66 × 106 Da). Compared to OPS, OPS-F had reduced levels of mannose, glucose, glucuronic acid and arabinose, but increased rhamnose, galacturonic acid and galactose, exhibiting enhanced solubility and lower apparent viscosity. Fourier transform infrared spectroscopy and nuclear magnetic resonance analysis showed minimal changes in polysaccharide structure post-fermentation. Moreover, despite a decrease in antioxidant activity post-fermentation, OPS-F exhibited superior immunomodulatory potential. In conclusion, fermenting okra juice with lactic acid bacteria alters the physicochemical properties of crude polysaccharides and enhances their immunomodulatory activity, offering a promising approach for developing new functional food resources.

Polysaccharides from red seaweeds: Effect of extraction methods on physicochemical characteristics and antioxidant activities

Seaweed polysaccharides are reported to possess biological and medicinal properties. Specifically, the antioxidant activity of seaweed extracts is the subject of intensive research due to ever-increasing demands from the food and pharmaceutical industries. The structure and composition of polysaccharides (carrageenan, xylan) extracted from four red seaweed species: Chondrus crispus (CC), Ahnfeltiopsis devoniensis (AD), Sarcodiotheca gaudichaudii (SG) and Palmaria palmata (PP) were compared and correlated with their antioxidant activity. Crude polysaccharide extracts were characterized by NMR (1H and 13C), HPLC-SEC, and FTIR spectroscopy. Total phenolic content (TPC), total sugars, sulfate and protein content were determined using spectrophotometric methods. The antioxidant capacity of each fractionated polysaccharide sample was evaluated by the following methods: DPPH (2,2-diphenyl-1-picrylhydrazyl radical), ABTS (2,2-azino-bis-[3-ethylbenzothiazoline-6-sulfonic acid]), superoxide dismutase (SOD), ferric-reducing power in FRAP (Ferric-reducing Antioxidant Power) assays, and the hydroxyl radical (OH) scavenging capacity assay. The highest antioxidant activity was found in extracts from A. devoniensis (ι/κ hybrid carrageenan), while the lowest was in P. palmata (xylans). Moreover, Chondrus crispus extracts did not possess SOD inhibition activity. Polysaccharides evaluated in this study offer high potential for applications in the food, pharmaceutical and biotechnology industries.

Impact of high pressure pre-treatment and hot water extraction on chemical properties of crude polysaccharide extract obtained from mushroom (Volvariella volvacea).

An examination of the process of extracting crude polysaccharides from Volvariella volvacea solely through hot water treatment (HWE) at 60, 80, and 100°C and through an approach involving high pressure processing (HPP) at 200, 400, and 600MPa followed by HWE. The physiological properties of the polysaccharides could be explained by the structural analysis performed via FT-IR spectroscopy and NMR spectroscopy, which revealed the extract composition of the protein-bound polysaccharides connected by β-glycosidic bonds. Under the extraction conditions investigated in this current study, the recommended extraction condition was a combination of HPP (600MPa, 10min) and HWE (60°C, 2h). This condition gave high crude polysaccharide yields (with a 2-12% increase), and β-glucan content (with a 15-20% increase) without disrupting the β-glycosidic bond, as compared to using HWE alone. High pressure extraction could be an alternative technique for reduced extraction temperatures of active compounds from mushrooms.

The Effect of G. applanatum Crude Polysaccharide Extract on Proinflammatory Cytokines and Proapoptotic Caspases in HeLa Cell Line: An In Vitro Study.

Polysaccharide extracts exhibit promise as potential anticancer agents. Among the fungi rich in polysaccharide content, G. applanatum stands out; however, its anticancer activity necessitates further investigation. This study aims to explore the impact of G. applanatum crude polysaccharide (GACP) extract by assessing its effects on cell viability, levels of proinflammatory cytokines such as TNF-α, IFN-γ, IL-2, and IL-12, and levels of proapoptotic markers including caspase-3 and caspase-9, as well as the percentages of necrosis and apoptosis in the HeLa cell line. Employing the HeLa cell line as a research model, four groups were studied: KN (media and DMSO), K+ (doxorubicin 10 μg/mL), P1 (G. applanatum extract 200 μg/mL), and P2 (G. applanatum extract 400 μg/mL). The G. applanatum extract was obtained via boiling distilled water. Anticancer activity was evaluated through the MTT test (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide) conducted over three treatment durations (24, 48, and 72 hours). Cytokine levels and caspase-3 and caspase-9 levels were assessed using the ELISA test. Cell apoptosis was determined using the Annexin V-PI biomarker and analyzed through flow cytometry. The MTT test exhibited optimal results at the 48-hour treatment mark. Cytokine level analysis revealed significant reductions in TNF-α, IFN-γ, IL-2, and IL-12 levels (p < 0.005). Concurrently, caspase-3 and caspase-9 levels exhibited substantial increases (p < 0.005). Flow cytometry highlighted the highest percentage of apoptosis in HeLa cells. In conclusion, G. applanatum's polysaccharide extract demonstrates potential as an anticancer and therapeutic agent for cancer treatment.

Potential Prebiotic Properties of Crude Polysaccharide Extract from Durian (Durio zibethinus Murr.) Seed Flour

The purpose of this research is to investigate the possible presence of prebiotic substances in durian seeds. The durian seed flour (DSF) was prepared by drum dry processing at 140 °C. The proximate composition of DSF 100 g contained moisture (2.43 g), protein (6.85 g), total fat (0.60 g), total carbohydrate (87.73 g), ash (2.39 g), and the sum of dietary fiber (4.48 g), respectively. The morphology of DSF was studied by SEM at a fixed magnification (100×). DSF appears to have smooth and rough surfaces, and the granules are covered by the gum. The optimal condition for extracting crude polysaccharide from DSF is ethanol 50 % at solvent ratios 1:8 at a temperature of 30 °C for 90 min, gave a maximum extraction yield of 5.28 % and the maximum amount of non-reducing sugar content at 392.19 ± 10.60 mg mL−1. The amount of non-reducing sugar is represented by an oligosaccharide referred to as a prebiotic. After being digested in vitro, the crude polysaccharide extract and inulin each had a total percentage of hydrolysis that was 14.68 and 11.54 %, respectively. Thus, approximately 85.32 % of crude polysaccharide extract was estimated to reach the colon. Moreover, the crude polysaccharide extract has enhanced activity values higher than inulin with 3 probiotic strains. The enhanced activity value increases when the extract concentration increases. The concentration of crude polysaccharide extract was 5.0 %w/v. Lactobacillus rhamnosus TBRC 374 had the highest enhanced activity at 582.67 %. In comparison, Lactobacillus casei BCC 13300 and Lactobacillus acidophilus ATCC 4356 with enhanced activity were 543.78 and 288.34 %, respectively. Significant advancements have been made in the utilization of plant waste and the development of novel prebiotic ingredients with the potential for application in food as a result of this research. HIGHLIGHTSDurian seed is one of the wastes that still has a good source of nutrition. Durian seed flour was prepared by drum dry processing that can maintain fiber and texture, making it suitable for application in the food industry. Moreover, the crude polysaccharide extract showed prebiotic properties. These are good prebiotic candidates that might be useful in producing functional foods. GRAPHICAL ABSTRACT

Optimisation of the extraction of crude polysaccharides from Clinacanthus nutans leaves for antioxidant applications: Content analysis, chemometrics and metabolomics analysis

This study aimed to evaluate the optimal method for extracting crude polysaccharides from Clinacanthus nutans leaves (CNP) as natural antioxidants. Five different extraction methods, including hot water (HW_CNP), ultrasound-assisted (UL_CNP), freeze-thaw (FT_CNP), alkaline (ALK_CNP), and acid (ACD_CNP). The extraction yield, chemical composition, antioxidant properties, and chemometrics and metabolomics analysis were investigated and compared. The results showed that the ALK_CNP and UL_CNP methods were the most effective for extracting CNP, with the highest extraction yield and antioxidant properties. The total phenolic content (TPC) was found to be directly related to antioxidant activity, with the highest TPC exhibited by the FT_CNP. The metabolomics analysis confirmed that the extraction method played a significant role in determining the antioxidant properties of CNP extracts. The findings of this study have implications for the food industry, as natural antioxidants are in high demand for food preservation and enhancement.

Physicochemical Properties, Antioxidant and Anti-Tyrosinase Activities of Durio zibethinus Murray and Value Added for Cosmetic Product Formulation

Durio zibethinus Murray, or durian fruit, is considered the “King of fruit” in various parts of Southeast Asia. It is classified in the Durio genus, which has been reported to be beneficial in the medical and pharmaceutical industry. The objective of this study was to investigate the physicochemical properties, biological activities, and cosmetic applications of the polysaccharide extractions from the green (GH) and white (WH) fruit-hulls and the ethanolic extracts from the pulp (P) and seeds (S) of D. Zibethinus. The extracts were developed as cosmetic products, and skin irritation was assessed by volunteers. P, S, GH, and WH extracts gave percentage yields of 12.13, 4.01, 1.60, and 1.18, respectively. The morphological structure of GH was highly porous, while the surface of WH was flattened, with no porosity observed. The seed extract showed the highest total phenolic content and antioxidant activity (DPPH) of 0.33 ± 0.01 mg GAE/g and IC50 0.08 ± 0.00 mg/mL, respectively, while no anti-tyrosinase activity was detected in any individual extract. The total phenolic content, DPPH assay, and anti-tyrosinase activity after blending the extracts were significantly (p &lt; 0.05) higher than in the individual extracts. Combining extracts such as SGH and SGWH gave IC50 values of anti-tyrosinase activity at 8.69 ± 1.82 mg/mL and 0.067 ± 0.00 mg/mL, respectively. No durian extracts exhibited growth inhibition against Staphylococcus aureus, Streptococcus pyogenes, and Cutibacterium acnes. A gel formulation containing durian extract was prepared by a cold process. It had good stability, with no skin irritation reported by the volunteers. Both crude durian and polysaccharide extracts showed promise as active ingredients in cosmetic products.

The Effect of Ganoderma applanatum Crude Polysaccharide Against a-Smooth Muscle Actin and Matrix Metalloproteinase-1 Expressions in Mice after Induced by Carbon Tetrachloride

Liver fibrosis was regarded as result of wound healing process in chronic liver injury. The α-smooth muscle actin (α-SMA) and matrix metalloproteinase-1 (MMP1) are several indicators for extracellular matrik (ECM) production which relate in fibrosis process. This study aims to determine the effect of Ganoderma applanatum crude polysaccharide (GACP) extract on carbon tetrachloride (CCl4)-against α-smooth muscle actin and matrix metalloproteinase expressions. A total of 24 male mice strain BALB/C, 3-4 months were divided into 6 groups. control group (distilled water and olive oil), model group (distilled water and CCl4), low G. applanatum crude polysaccharide group (G. applanatum extract at the dose of 25 mg/kg BW and CCl4), middle G. applanatum crude polysaccharide group (G. applanatum extract at the dose of 50 mg/kg b.w and CCl4), high G. applanatum crude polysaccharide group (G. applanatum extract at the dose of 100 mg/kg BW and CCl4) and silymarin group (silymarin 100 mg/kg BW and CCl4). Expressions of α-SMA and MMP1 were examined by immunohistochemical methods. The observation of immunohistochemistry used an inverted microscope at 200× magnification. The results of the observation were analyzed using ImageJ software to measure the percentage of α-SMA and MMP1 expressions. All of the data were analyzed by GraphPad Prism Software. The results showed that G. applanatum extracts prevented a significant increase in α-SMA expression and decrease MMP1 expression (p &lt; 0.05) in comparison with the model group. Therefore, G. applanatum extracts play as liver protector against chronic liver injury after successfully inhibit α-SMA expression and prevent reduction of MMP1 expression.

Microalga Chlorella sp. extract induced apoptotic cell death of cholangiocarcinoma via AKT/mTOR signaling pathway

Cancer is the leading cause of death worldwide. Drug resistance and relapse after current standard treatments frequently occur; thus, alternative and effective treatments are required. Algae and cyanobacteria are abundant organisms that serve as bioresources of nutrients/metabolites, which are attractive sources of numerous bioactive compounds for drug discovery. In the present study, we, therefore, investigated anti-cancer activities of crude polysaccharide and ethanolic extracts from Chlorella sp., Sargassum spp., and Spirulina sp. against cell lines of five top-leading cancers including lung cancer (A549), cervical cancer (Hela), breast cancer (MCF7), hepatocellular carcinoma (Huh7), and cholangiocarcinoma (CCA; KKU213A). Only ethanolic extracts of Chlorella sp. showed consistent inhibition of growth of all cancer cell types. CCA was the most sensitive to Chlorella sp. ethanolic extract with CC50 of 277.4, 400.5, and 313.4 µg/mL for KKU055, KKU100, and KKU213A cells, respectively. Flow cytometric analysis demonstrated that CCA cell death was triggered via apoptosis pathway in accompany with lowering procaspase-3, -8, and -9 and increasing caspase enzymatic activity in addition to reducing anti-apoptosis Bcl-2 protein. Interestingly, the treatment of the extract at 400 µg/mL greatly inhibited the AKT/mTOR survival signaling as evidenced by significant reduction of phosphorylated-AKT and phosphorylated-mTOR proteins. The presence of reported bioactive compounds, gallic acid, and lutein, were confirmed in Chlorella sp. extract by high-performance liquid chromatography. Gallic acid and lutein treatment caused a significant reduction of KKU055, KKU100, and KKU213A cell viability. This study demonstrated the anti-cancer effect of Chlorella sp. ethanolic extract to promote cancer cell death via inhibition of AKT/mTOR pathway.

Anticancer and Antioxidant Activity of Water-Soluble Polysaccharides from Ganoderma aff. australe against Human Osteosarcoma Cells.

Wild mushrooms have gained great importance for being a source of biologically active compounds. In this work, we evaluate the anticancer and antioxidant activity of a water-soluble crude polysaccharide extract isolated from the fruiting bodies of the Ganoderma aff. australe (GACP). This mushroom was collected in San Mateo (Boyacá, Colombia) and identified based on macroscopic and microscopic characterization. GACP was characterized by UV-Vis spectroscopy, Fourier-transform infrared spectroscopy, high-performance liquid chromatography-diode array detector, and nuclear magnetic resonance. The antiradical and antioxidant activity were evaluated by different methods and its anticancer activity was verified in the osteosarcoma MG-63 human cell line. Chemical and spectroscopic analysis indicated that GACP consisted of β-D-Glcp-(1→, →3)-β-D-Glcp-(1→ and α-D-Glcp-(1→ residues. The results of the biological activity showed that GACP exhibited high antioxidant activity in the different methods and models studied. Moreover, the results showed that GACP impaired cell viability (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay) and cell proliferation (clonogenic assay) in a dose-response manner on MG-63 cells. The findings of this work promote the use of mushroom-derived compounds as anticancer and antioxidant agents for potential use in the pharmaceutical and food industries.

Ultrasonic-Cellulase Synergistic Extraction of Crude Polysaccharides from Moringa oleifera Leaves and Alleviation of Insulin Resistance in HepG2 Cells.

Moringa oleifera leaves (MOL) are a new food resource, rich in functional factors. MOL polysaccharides are important active macromolecules within MOL. However, there are problems, such as low extraction rates and lack of evidence for functional activity. Therefore, in this experiment, single-factor experiments were carried out using MOL powder as the raw material, and the Plackett–Burman test was used to screen the significantly influential test factors. The extraction process of MOL polysaccharide was optimized by response surface methodology. The insulin resistance alleviating activity of MOLP polysaccharides was initially explored. The results showed that the extraction of Moringa oleifera leaves crude polysaccharides (MOLP) by ultrasonic assisted cellulase enzymatic digestion was (17.03 ± 1.03)%, and the obtained MOLP was a crude polysaccharide with an average molecular weight (Mw) of 279.48 kDa, consisting of fucose, rhamnose, arabinose, galactose, glucose, xylose, mannose, galacturonic acid, and glucuronic acid. MOLP had an IC50 value of 8.02 mg/mL for α-glucosidase and scavenging activity against free radicals such as ABTS, DPPH, hydroxyl radicals, and superoxide anion with an IC50 value of 0.21 mg/mL 0.31 mg/mL 0.97 mg/mL 0.49 mg/mL. At the same time, MOLP significantly enhanced the glucose consumption, glycogen synthesis, CAT, SOD, GSH-Px activity, and reduced the MDA and ROS content in high glucose-induced insulin-resistant HepG2 (IR-HepG2) cells. This experiment improved the extraction rate of MOLP and demonstrated that MOLP has antioxidant activity and α-glucosidase inhibitory activity, which can alleviate the insulin resistance of high glucose-induced HepG2 cells. It provides partial data support for the possible hypoglycemic effect of MOLP by alleviating oxidative stress, and also provides new ideas for the in-depth study of basic research and industrial application of MOLP.

Pressurized hot water extraction of crude polysaccharides, β-glucan, and phenolic compounds from dried gray oyster mushroom

This study aimed to investigate the effects of temperature (100 °C–140 °C), pressure (0.4–1.0 MPa), and extraction time (20–60 min) on the extraction yield of crude polysaccharides, β-glucan, and total phenolic compounds with pressurized hot water extraction of gray oyster mushrooms (Pleurotus sajor-caju (Fr.) Singer). The extraction conditions were optimized by maximizing all responses, using a central composite design (CCD)-based response surface methodology. Temperature mainly affected the increase in the extracted yield and phenolic content. However, exceeded temperature and extraction time negatively affected the β-glucan yield. The optimal extraction conditions were at 140 °C, 0.92 MPa, and 40 min with a corresponding extraction yield of 3.20 ± 0.17 g/100 g, as well as β-glucan and total phenolic contents of 43.84 ± 3.86 mg/100 g and 8.49 ± 0.66 mg gallic acid equivalent/100 g dried extract, respectively. The antioxidant activities of extracts were estimated by in vitro biological assays. The SC50 values of DPPH, NO, and ABTS assays were 1.66 ± 0.02, 1.50 ± 0.19, and 6.94 ± 1.04 mg/mL, respectively. This work demonstrates an alternative approach to valorize over-produced mushrooms.