Pro-Apoptotic Activity and Cell Cycle Arrest of Caulerpa sertularioides against SKLU-1 Cancer Cell in 2D and 3D Cultures.

Cancer is a disease with the highest mortality and morbidity rate worldwide. First-line drugs induce several side effects that drastically reduce the quality of life of people with this disease. Finding molecules to prevent it or generate less aggressiveness or no side effects is significant to counteract this problem. Therefore, this work searched for bioactive compounds of marine macroalgae as an alternative treatment. An 80% ethanol extract of dried Caulerpa sertularioides (CSE) was analyzed by HPLS-MS to identify the chemical components. CSE was utilized through a comparative 2D versus 3D culture model. Cisplatin (Cis) was used as a standard drug. The effects on cell viability, apoptosis, cell cycle, and tumor invasion were evaluated. The IC50 of CSE for the 2D model was 80.28 µg/mL versus 530 µg/mL for the 3D model after 24 h of treatment exposure. These results confirmed that the 3D model is more resistant to treatments and complex than the 2D model. CSE generated a loss of mitochondrial membrane potential, induced apoptosis by extrinsic and intrinsic pathways, upregulated caspases-3 and -7, and significantly decreased tumor invasion of a 3D SKLU-1 lung adenocarcinoma cell line. CSE generates biochemical and morphological changes in the plasma membrane and causes cell cycle arrest at the S and G2/M phases. These findings conclude that C. sertularioides is a potential candidate for alternative treatment against lung cancer. This work reinforced the use of complex models for drug screening and suggested using CSE's primary component, caulerpin, to determine its effect and mechanism of action on SKLU-1 in the future. A multi-approach with molecular and histological analysis and combination with first-line drugs must be included.

Ulvophyte Green Algae Caulerpa lentillifera: Metabolites Profile and Antioxidant, Anticancer, Anti-Obesity, and In Vitro Cytotoxicity Properties.

Marine algae have excellent bioresource properties with potential nutritional and bioactive therapeutic benefits, but studies regarding Caulerpa lentillifera are limited. This study aims to explore the metabolites profile and the antioxidant, anticancer, anti-obesity, and in vitro cytotoxicity properties of fractionated ethanolic extract of C. lentillifera using two maceration and soxhlet extraction methods. Dried simplicia of C. lentillifera was mashed and extracted in ethanol solvent, concentrated and evaporated, then sequentially partitioned with equal volumes of ethyl acetate and n-Hexane. Six samples were used in this study, consisting of ME (Maceration-Ethanol), MEA (Maceration-Ethyl Acetate), MH (Maceration-n-Hexane), SE (Soxhletation-Ethanol), SEA (Soxhletation-Ethyl Acetate), and SH (Soxhletation-n-Hexane). Non-targeted metabolomic profiling was determined using LC-HRMS, while antioxidant, anti-obesity, and anticancer cytotoxicity were determined using DPPH and ABTS, lipase inhibition, and MTT assay, respectively. This study demonstrates that C. lentillifera has several functional metabolites, antioxidant capacity (EC50 MH is very close to EC50 of Trolox), as well as anti-obesity properties (EC50 MH < EC50 orlistat, an inhibitor of lipid hydrolyzing enzymes), which are useful as precursors for new therapeutic approaches in improving obesity-related diseases. More interestingly, ME, MH, and SE are novel bioresource agents for anticancer drugs, especially for hepatoma, breast, colorectal, and leukemia cancers. Finally, C. lentillifera can be a nutraceutical with great therapeutic benefits.

Dietary Supplementation of Caulerpa racemosa Ameliorates Cardiometabolic Syndrome via Regulation of PRMT-1/DDAH/ADMA Pathway and Gut Microbiome in Mice.

This study evaluated the effects of an aqueous extract of Caulerpa racemosa (AEC) on cardiometabolic syndrome markers, and the modulation of the gut microbiome in mice administered a cholesterol- and fat-enriched diet (CFED). Four groups of mice received different treatments: normal diet, CFED, and CFED added with AEC extract at 65 and 130 mg/kg body weight (BW). The effective concentration (EC50) values of AEC for 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and lipase inhibition were lower than those of the controls in vitro. In the mice model, the administration of high-dose AEC showed improved lipid and blood glucose profiles and a reduction in endothelial dysfunction markers (PRMT-1 and ADMA). Furthermore, a correlation between specific gut microbiomes and biomarkers associated with cardiometabolic diseases was also observed. In vitro studies highlighted the antioxidant properties of AEC, while in vivo data demonstrated that AEC plays a role in the management of cardiometabolic syndrome via regulation of oxidative stress, inflammation, endothelial function (PRMT-1/DDAH/ADMA pathway), and gut microbiota.

Supplement of Caulerpa lentillifera polysaccharide by pre-prandial gavage and free feeding demonstrates differences to prevent obesity and gut microbiota disturbance in mice.

BACKGROUND: Caulerpa lentillifera has received extensive attention regarding expansion of its farming and increasing consumption. In our previous study, the structure of C. lentillifera polysaccharide (CLP) was elucidated. However, little information is available about its health effects. In this study, the anti-obesity effect of CLP was investigated by using a high-fat diet-induced obese mice model with two different supplementation methods. RESULTS: In vitro simulated digestion results showed that CLP significantly decreased the lipid digestibility and induced the lipid droplets aggregation in the intestinal stage to inhibit the absorption of lipids. As revealed by 16S ribosomal RNA sequencing and non-targeted metabolomics, supplement of CLP by both pre-prandial gavage and free feeding patterns effectively prevented mice obesity via ameliorating intestinal flora disturbance and regulating bile acids circulation metabolism. Of note was that CLP administration had no effect on short-chain fatty acids production, suggesting the anti-obesity effect was uncorrelated with their production. Moreover, pre-prandial administration of CLP had a better anti-obesity effect in lowering body weight and serum lipid levels, but the free feeding resulted in a higher α-diversity of gut microbiota. CONCLUSION: The findings of this study indicate that CLP could be a potential anti-obesity nutraceutical and that pre-prandial supplement of CLP may be a better intake method to exhibit its hypolipidemic effect. © 2022 Society of Chemical Industry.

An Efficient and Quick Analytical Method for the Quantification of an Algal Alkaloid Caulerpin Showed In-Vitro Anticancer Activity against Colorectal Cancer.

Biological invasion is the successful spread and establishment of a species in a novel environment that adversely affects the biodiversity, ecology, and economy. Both invasive and non-invasive species of the Caulerpa genus secrete more than thirty different secondary metabolites. Caulerpin is one of the most common secondary metabolites in genus Caulerpa. In this study, caulerpin found in invasive Caulerpa cylindracea and non-invasive Caulerpa lentillifera extracts were analyzed, quantified, and compared using high-performance thin layer chromatography (HPTLC) for the first time. The anticancer activities of caulerpin against HCT-116 and HT-29 colorectal cancer (CRC) cell lines were also tested. Caulerpin levels were found higher in the invasive form (108.83 ± 5.07 µg mL-1 and 96.49 ± 4.54 µg mL-1). Furthermore, caulerpin isolated from invasive Caulerpa decreased cell viability in a concentration-dependent manner (IC50 values were found between 119 and 179 µM), inhibited invasion-migration, and induced apoptosis in CRC cells. In comparison, no cytotoxic effects on the normal cell lines (HDF and NIH-3T3) were observed. In conclusion, HPTLC is a quick and novel method to investigate the caulerpin levels found in Caulerpa extracts, and this paper proposes an alternative utilization method for invasive C. cylindracea due to significant caulerpin content compared to non-invasive C. lentillifera.

Dietary fibers obtained from Caulerpa lentillifera prevent high-fat diet-induced obesity in mice by regulating the gut microbiota and metabolite profiles.

Due to its expanded farming and growing consumption, Caulerpa lentillifera has received extensive attention. In the present study, the physicochemical properties of insoluble dietary fibers from C. lentillifera (CL-IDFs) were evaluated in vitro, and 16S rRNA sequencing analysis as well as non-targeted metabolomics were performed to investigate the hypolipidemic effects of CL-IDFs and their combined supplementation (CL-TDFs, composed of CL-IDFs and soluble polysaccharides of C. lentillifera) in vivo. The results show that CL-IDFs exhibited superior physicochemical capacities on the binding of water, oil, and glucose. In addition, CL-IDF and CL-TDF administration could regulate the gut microbiota, increase acetic and propionic acid levels, and restore the metabolic disorders of amino, fatty, and bile acids in obese mice. Notably, considering the processing cost of C. lentillifera and the equal anti-obesity effects of CL-IDFs and CL-TDFs, fresh whole-food supplementation of C. lentillifera may be a cost-effective way to prevent obesity.

Phytochemical Analysis and Molecular Identification of Green Macroalgae Caulerpa spp. from Bali, Indonesia.

The studies of the Bulung Boni and Bulung Anggur (Caulerpa spp.) species and secondary metabolites are still very limited. Proper identification will support various aspects, such as cultivation, utilization, and economic interests. Moreover, understanding the secondary metabolites will assist in developing algae-based products. This study aimed to identify these indigenous Caulerpa algae and analyze their bioactive components. The tufA sequence was employed as a molecular marker in DNA barcoding, and its bioactive components were identified using the GC-MS method. The phylogenetic tree was generated in MEGA 11 using the maximum likelihood method, and the robustness of the tree was evaluated using bootstrapping with 1000 replicates. This study revealed that Bulung Boni is strongly connected to Caulerpa cylindracea. However, Bulung Anggur shows no close relationship to other Caulerpa species. GC-MS analysis of ethanolic extracts of Bulung Boni and Bulung Anggur showed the presence of 11 and 13 compounds, respectively. The majority of the compounds found in these algae have been shown to possess biological properties, such as antioxidant, antibacterial, anticancer, anti-inflammation, and antidiabetic. Further study is necessary to compare the data obtained using different molecular markers in DNA barcoding, and to elucidate other undisclosed compounds in these Caulerpa algae.

Structural characterization and SARS-CoV-2 inhibitory activity of a sulfated polysaccharide from Caulerpa lentillifera.

Caulerpa lentillifera (Bryopsidophyceae, Chlorophyta) is an edible seaweed attracting great attention for its expansion of farming scale and increasing consumption in these years. In the present study, a sulfated polysaccharide (CLSP-2) was isolated and separated from C. lentillifera, and its chemical structure was elucidated by a series of chemical and spectroscopic methods. Among these methods, mild acid hydrolysis and photocatalytic degradation were applied to release mono- and oligo-saccharide fragments which were further identified by HPLC-MSn analysis, affording the information of the sugar sequences and the sulfate substitution in CLSP-2. Results indicated that the backbone of CLSP-2 was constructed of →6)-ß-Manp-(1→ with sulfated branches at C2, which were comprised of prevalent →3)-ß-Galp4S-(1→, →3)-ß-Galp2,4S-(1→, and minor Xyl. In addition, the virus neutralization assay revealed that CLSP-2 could effectively protect HeLa cells against SARS-CoV-2 infection with an IC50 of 48.48 µg/mL. Hence, the present study suggests CLSP-2 as a promising agent against SARS-CoV-2.

Microwave irradiation is a suitable method for caulerpin extraction from the green algae Caulerpa racemosa (Chlorophyta, Caulerpaceae).

Caulerpin is a bisindolic alkaloid that has been obtained from many species of the genus Caulerpa. The main objective of this paper is to evaluate four extraction methods of caulerpin in the C. racemosa: maceration (DMA), Soxhlet extraction (SOX), ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE). The methods were compared through caulerpin content quantified by Ultraviolet-visible (UV-vis) spectrophotometry. The highest extract yield was obtained by SOX but the highest contain of caulerpin was presented in the MAE extract. The caulerpin content was significant different within the extacts by MAE and UAE, it yielded by MAE more than three times as much as UAE. The most efficient caulerpin extraction method had the parameters solvent, temperature and time optimised. Thus, the best conditions were achieved with MAE in ethanol during 7 min at 90 °C. Therefore, this work suggests an improved routine analysis of caulerpin by the green chemistry concept.

The Enhancing Immune Response and Anti-Inflammatory Effects of Caulerpa lentillifera Extract in RAW 264.7 Cells.

BACKGROUND: Caulerpa lentillifera (CL) is a green seaweed, and its edible part represents added value as a functional ingredient. CL was dried and extracted for the determination of its active compounds and the evaluation of its biological activities. The major constituents of CL extract (CLE), including tannic acid, catechin, rutin, and isoquercetin, exhibited beneficial effects, such as antioxidant activity, anti-diabetic activity, immunomodulatory effects, and anti-cancer activities in in vitro and in vivo models. Whether CLE has an anti-inflammatory effect and immune response remains unclear. METHODS: This study examined the effect of CLE on the inflammatory status and immune response of lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and the mechanisms involved therein. RAW264.7 cells were treated with different concentrations of CLE (0.1-1000 µg/mL) with or without LPS (1 µg/mL) for 24 h. Expression and production of the inflammatory cytokines, enzymes, and mediators were evaluated. RESULTS: CLE suppressed expression and production of the pro-inflammatory cytokines IL-6 and TNF-α. Moreover, CLE inhibited expression and secretion of the inflammatory enzyme COX-2 and the mediators PGE2 and NO. CLE also reduced DNA damage. Furthermore, CLE stimulated the immune response by modulating the cell cycle regulators p27, p53, cyclin D2, and cyclin E2. CONCLUSIONS: CLE inhibits inflammatory responses in LPS-activated macrophages by downregulating inflammatory cytokines and mediators. Furthermore, CLE has an immunomodulatory effect by modulating cell cycle regulators.

Immunomodulatory sulfated polysaccharides from Caulerpa racemosa var. peltata induces metabolic shifts in NF-κB signaling pathway in RAW 264.7 macrophages.

Algal polysaccharide activates macrophages to alter physiologic biomarkers to drive the immunomodulatory phenotype, but it lacks specific biomarkers involved in the biochemical underpinning process. Here, we undertook an extensive analysis of the RAW 264.7 macrophages induced by an immunostimulating sulfated polysaccharide from Caulerpa racemosa var. peltata (CRVP-1) employing combined transcriptomic, proteomic, and metabolomic analyses to reveal the molecular details occurring in the CRVP-1-induced immunomodulatory process. The omics profiling of CRVP-1-activated macrophage demonstrated a total of 8844 genes (4354 downregulated and 4490 upregulated), 1243 proteins (620 downregulated and 623 upregulated), and 68 metabolites (52 downregulated and16 upregulated). Further, the co-mapped correlation network of omics combined with Western blot and immunofluorescence staining indicated that the cluster of differentiation 14 (CD14) might assist Toll-like receptor 4 (TLR4) involved in nuclear factor kappa-B (NF-κB) signaling pathway to drive the immunomodulatory phenotype. Together, our results discover novel physiologic biomarkers in the immunomodulatory activities of algal polysaccharides.

In Vitro Antitumor Potential of Sulfated Polysaccharides from Seaweed Caulerpa cupressoides var. flabellata.

Seaweeds are important source of bioactive compounds, including sulfated polysaccharides (SP). Because of their structural heterogeneity, these compounds are promising sources of anticancer compounds. SP from brown and red seaweeds have shown antimelanoma activity in different in vitro and in vivo models. However, SP from green seaweed are still poorly evaluated. Therefore, SP were extracted from the green alga Caulerpa cupressoides var. flabellata, and their antiproliferative, anti-migratory, and inhibitory effect on melanin production on B16-F10 melanoma cells was evaluated. Cell assays, including flow cytometry, demonstrated that SP (100-1000 µg mL-1) are non-cytotoxic, do not induce apoptosis or necrosis, and do not interfere with cell cycle. However, SP (1000 µg mL-1) were found to significantly inhibit cell colony formation (80-90%), cell migration (40-75%), and melanin production (~ 20%). In summary, these results showed that SP inhibited important melanoma development events without cytotoxicity effects, suggesting that C. cupressoides may be an important source of SP with antitumor properties.

Anti-proliferative and ROS-inhibitory activities reveal the anticancer potential of Caulerpa species.

Seaweeds are considered a promising functional food and safe for human consumption as they have significant health benefits. Five abundant tropical seaweeds, Caulerpa racemosa var. macrophysa, Caulerpa scalpelliformis, Grateloupia indica, Sargassum linearifolium, and Spatoglossum asperum rich in metabolites, phenolic, and flavonoid compounds, were analyzed for the anti-proliferative and ROS inhibitory activities including transcript expression of cancer-linked key genes and apoptosis. C. racemosa var. macrophysa showed the maximum effective activities with a lower dose of extract, about 130 ± 30 and 23 ± 1 µg ml-1 EC50 dose for HeLa and Huh-7, respectively, followed by C. scalpelliformis, showing EC50 dose about 200 ± 10 and 140 ± 30 µg ml-1, respectively. Similarly, about 56% and 54% ROS inhibition were determined with Caulerpa spp. for HeLa and Huh-7 cells, respectively. Results indicated that tropical green seaweed Caulerpa spp. (C. racemosa var. macrophysa and C. scalpelliformis) have substantial potential of ROS inhibition. Further, it was observed that different cancer-linked marker proteins encoding genes were deferentially expressed with seaweed extracts in different cell lines. Overall, it is concluded that Caulerpa spp. are rich in antioxidant and anti-proliferative activities. Caulerpa spp. have potential to be explored further for cancer preventive properties or functional food or nutraceuticals applications.

An Analysis of the Nutritional and Health Values of Caulerpa racemosa (Forsskål) and Ulva fasciata (Delile)-Two Chlorophyta Collected from the Philippines.

Polysaccharides, lipids and amino acid profiles were investigated to understand the nutritional value of Caulerpa racemosa and Ulva fasciata from the Philippines. The results revealed that both species contain high amounts of proteins (8.8-19.9% for C. racemosa and 8.0-11.1% for U. fasciata). The portions of the total amino acids that were essential amino acids (EAAs) (45.28 ± 0.12% for C. racemosa and 42.17 ± 0.12% for U. fasciata) out were comparable to FAO/WHO requirements. Leucine, valine, isoleucine, and lysine are the dominant EAAs in C. racemosa, while leucine, valine, lysine, and phenylalanine are those in U. fasciata. The fatty acid profiles are dominated by monounsaturated fatty acids and polyunsaturated fatty acids in C. racemosa (56.2%), while saturated fatty acids (72.1%) are dominant in U. fasciata. High C18/C20 polyunsaturated fatty acid ratios were recorded in both species. Mineral contents for both seaweeds were within levels considered safe for functional foods. Total pigment content of C. racemosa (140.84 mg/g dw) was almost 20 times higher than that of U. fasciata (7.54 mg/g dw). Hot water extract (HWE) from C. racemosa showed in vitro antiherpetic activity without cytotoxicity. Nutritional characteristics confirmed that C. racemosa could be potentially used as a nutritious and functional food items for human consumption.

The potential of polysaccharide extracts from Caulerpa lentillifera waste.

Caulerpa is a marine macroalgae and is rich in polysaccharides, which have the potential for immunostimulatory and anticoagulant activity. The objective of this work was to increase the value of C. lentillifera waste by polysaccharide extraction. A polysaccharide yield of about 25% of dry weight was obtained under the following optimized conditions: two-stage extraction (60 min/stage) using a ratio of 1:15 (w/v) at 90 °C, and 2× precipitation by the final concentration of 75% ethanol. The polysaccharide extracts contained a non-reducing sugar that accounted for 44% of weight extracts as a major sugar and consisted of four neutral sugars: mannose (33.3%), galactose (31.9%,), glucose (27.0%) and xylose (7.6%). In addition, it contained sulfate, which is approximately 8.37% of weight extracts and had a phenolic content of around 1.27 mg GAE/g sample. Moreover, it demonstrated α-glucosidase inhibitory activity with an IC50 value of 13.59 mg/mL. This result suggests that the polysaccharide extracts could potentially be used for preventing diabetes disease. The economic analysis also showed an economic feasibility for producing polysaccharide extracts from C. lentillifera waste. This is an alternative for farmers in order to increase the value of C. lentillifera waste.

Three New Butenolides from the Green Alga Caulerpa racemosa var. turbinata.

Three new butenolides, caulerpalide A and a pair of enantiomers, (+)-caulerpalide B and (-)-caulerpalide B, together with seven known compounds, have been isolated from the green alga Caulerpa racemosa var. turbinata. All these structures were determined by spectroscopic techniques. The absolute configurations of caulerpalide A, (+)-caulerpalide B and (-)-caulerpalide B were elucidated by the method of ECD calculation. This is the first separation of butenolides from the algae of genus Caulerpa. Additionally, the antibacterial activities of the nine isolated compounds were also evaluated.

Caulerpa okamurae extract attenuates inflammatory interaction, regulates glucose metabolism and increases insulin sensitivity in 3T3-L1 adipocytes and RAW 264.7 macrophages.

OBJECTIVE: To examine whether Caulerpa okamurae ethanolic extract (COE) could inhibit obesity-mediated inflammation, improve glucose metabolism and increase insulin sensitivity, using in vitro cell models of RAW 264.7 macrophages and 3T3-L1 adipocytes. METHODS: We cocultured 3T3-L1 adipocytes in direct contact with lipopolysaccharide-stimulated RAW 264.7 macrophages and induced insulin resistance in 3T3-L1 adipocytes with tumor necrosis factor-α (TNF-α) in the presence or absence of 250 µg/mL of COE. We investigated various markers of inflammation, glucose regulation and insulin sensitivity in these models using Griess reagent to measure nitric oxide (NO) production, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxyglucose to measure glucose uptake, Western blot analysis to quantify protein expression and reverse transcriptase-polymerase chain reaction to evaluate mRNA expression. RESULTS: We found that COE (250 µg/mL) significantly inhibited the lipopolysaccharide-induced inflammatory response in RAW 264.7 macrophages by downregulating NO production, nitric oxide synthase 2 expression and nuclear translocation of nuclear factor-κB. COE also showed similar anti-inflammatory activity in coculture, along with decreased TNF-α, interleukin-6 and monocyte chemoattractant protein mRNA expression. In addition, COE also improved glucose uptake in coculture by upregulating glucose transporter-4 (GLUT-4) and adiponectin and reducing serine phosphorylation of insulin receptor substrate-1 (IRS1). In the TNF-α-induced insulin resistance model of 3T3-L1 adipocytes, COE significantly improved both basal and insulin-stimulated glucose uptake, accompanied by phosphorylation of IRS1 at tyrosine 632, phospho-5' adenosine monophosphate-activated protein kinase α and glycogen synthase kinase-3ß (Ser9) as well as upregulation of GLUT-4. CONCLUSION: Together, these findings suggest that COE has potential to treat or prevent obesity-induced metabolic disorders.

Physicochemical characterization and bile acid-binding capacity of water-extract polysaccharides fractionated by stepwise ethanol precipitation from Caulerpa lentillifera.

Herein, water-extracted Caulerpa lentillifera polysaccharides were structurally characterized and their bile acid-binding capacities were investigated. WCLP-25, WCLP-40, WCLP-55, WCLP-70, and WCLP-85 were obtained by graded ethanol precipitation with ethanol concentrations of 25%, 40%, 55%, 70%, and 85%. The total carbohydrate, protein, uronic acid and sulfate contents as well as the monosaccharide composition, molecular weight, and rheological properties were determined. Their infrared spectra, thermogravimetric curves, and scanning electron microscopy (SEM) images were acquired. The hypolipidaemic effects of the WCLPs were assessed with in vitro simulated bile acid-binding capacity experiments. The WCLPs are high-molecular-weight sulfated heteropolysaccharides, and the ethanol concentration significantly influenced the physicochemical properties of the extract. The bile acid-binding capacities of WCLP-55 and WCLP-70 were significantly higher than those of the other tested WCLP samples, which may be due to their higher neutral sugar, uronic acid and sulfate contents or due to their higher viscosities and a larger sheet structure based on SEM. This study will broaden the sources of raw materials for functional foods and provide a reference for the scientific use of C. lentillifera.

Anti-inflammatory activity and structural identification of a sulfated polysaccharide CLGP4 from Caulerpa lentillifera.

In the present study, the in vitro anti-inflammatory activity of four purified polysaccharides (CLGP1, CLGP2, CLGP3 and CLGP4) extracted from edible green algae Caulerpa lentillifera was evaluated. As a result, CLGP4 exhibited more effectively inhibitory effect on LPS-induced HT29 cells, including reducing the production of IL-1ß, TNF-α, SIgA and mucin2, and decreasing the expression of IL-1ß and TNF-α. According to the results, CLGP4 showed a better anti-inflammatory effect, might highly related to the presence of sulfate groups. Furthermore, the structure of CLGP4 was analyzed by methylation analysis, GC-MS and NMR spectroscopy. It was found that CLGP4 was a novel xylogalactomanan consisting of ß-(1 â†’ 4)-Manp, →2,4)Manp(1→, ß-(1 â†’ 2)-Manp, ß-(1 â†’ 3)-Galp, ß-(1 â†’ 4)-Xylp, terminal ß-Galp and terminal ß-Xylp residues. Additionally, the sulfate groups were located on C-3 of â†’4)Xylp(1→, C-6 of â†’3)Galp(1→ and C-3 of â†’2)Manp(1→. These results could enlarge the potential application of CLGP4 as functional ingredient to attenuate inflammation.

Study on immunostimulatory activity and extraction process optimization of polysaccharides from Caulerpa lentillifera.

The process of extracting polysaccharides from the green algae Caulerpa lentillifera was studied by single factor experiments and response surface methodology. Additionally, the immunostimulatory activity of Caulerpa lentillifera polysaccharides (CLP) on RAW264.7 mouse macrophage was evaluated by in vitro cell experiments. The results showed that the optimal extraction conditions consisted of ultrasonification for 30 min, extraction time of 9 h, extraction temperature of 100 °C, and a ratio of water to raw material of 40:1. RAW264.7 macrophage exhibited enhanced phagocytosis with no toxic effects after treatment with CLP. In addition, CLP effectively increased the synthesis and secretion of cytokines (IL-6, TNF-α, IL-1ß, and NO), whereby the secretion levels of IL-6, TNF-α, and IL-1ß were 1,840.32 ± 21.03 pg/mL (50 µg/mL), 197.17 ± 3.13 ng/mL (50 µg/mL), and 1,178.35 ± 78.82 pg/mL (25 µg/mL), respectively. The polysaccharides contained in Caulerpa lentillifera have potential value for further development due to their immunological activity.