First Report of Sphingobacterium multivorum Causing Bacterial Rot of Cauliflower mushroom (Sparassis latifolia) in China.

Cauliflower mushroom (Sparassis latifolia), is widely distributed in Australia, North America, Europe, and East Asia (Bashir et al., 2020). It is known for its medicinal significance due to the availability of various pharmacological substances and their use in health supplements (Bashir et al., 2017). In recent years, with the development of artificial cultivation technology, S. latifolia has been industrialized in China, with an annual output value 50 million dollars. In March 2023, approximately 15% of S. latifolia showed obvious bacterial rot in mushroom hothouse (about 0.05 ha), located in Shuangliu county, Sichuan province, China (104°7'51"N, 30°25'2"E). The affected parts appear water-soaked, and become sunken and softened as the disease progresses. In the finally, all the fruiting body tissues turn into paste, with colors pale yellow, and have a foul smell. The pathogen was isolated from the margin of the lesions by dilution and streaking techniques onto Nutrient Agar, and incubated at 28℃ in the dark for 2-3 days. A single colony was re-streak for purification. Eight isolates were obtained from five samples collected randomly. The representative three isolates were selected for further characterization. For pathogenicity testing, ten health fruit bodies of S. latifolia were selected (for per isolate). Bacterial suspensions (1 × 107 CFU/ml) of the three isolates were applied to the fruiting body until wet, sterile water was used as controls. All the S. latifolia were maintained at 19±1℃, 85-100% relative humidity, and 18 h of light in the mushroom hothouse. Three days later, the inoculated fruiting bodies developed yellow color, and appear water-soaked, five days later, fruiting body gradually turn to soft and part turn to rot, seven days later, the fruiting body tissues completely turn into paste with a foul smell. The symptoms exhibited were similar to those of the original diseased fruiting bodies, while the control group remained healthy. The same bacterial were re-isolated from the infected fruiting bodies and subsequently identified by morphological characteristics and DNA sequenced. The pathogenicity test was conducted three times, each yielding similar results. The colonies of the pathogen are gram-negative rods, medium sized, convex, smooth, opaque, turning yellow after several days at a temperature 28℃. For molecular identification, the DNA of the representative three isolates was extracted using a Bacterial Genomic DNA Extraction Kit (Solarbio, Beijing). The 16S rRNA genes were amplified and sequenced with the primer 27F/1492R (Lane et al., 1985). Finally, the sequences were identical. The generated representative sequence was deposited in GenBank with accession number OR399122. BLASTn analysis showed 100% identity (1404/1404 bp) with previously deposited sequence (accession number CP068224) of S. multivorum FDAARGOS in GenBank. Based on the maximum likelihood method, phylogenetic analysis revealed 100% bootstrap support values with S. multivorum. Finally, the bacterium was identified as S. multivorum. This is the first report of S. multivorum causing bacterial rot of mushroom. The fruiting body of S. multivorum consists of multiple folded flat lobes, which are thin and have large surface area, may facilitate the infection of S. multivorum. Sphingobacterium sp. are named for their synthesize sphingolipids, which play an important role in bacterial infection (Kunz et al., 2019). These results will contribute to developing control strategies for this disease.

Potential osteoporosis-blocker Sparassis crispa polysaccharide: Isolation, purification and structure elucidation.

This study aimed to investigate the structural characterization of water-soluble polysaccharides from Sparassis crispa and their effects on the proliferation and differentiation of mouse osteoblasts. Three fractions (F-1, F-2, and F-3) were obtained from crude polysaccharides by a DEAE-52 cellulose column. The main fraction (F-1) was further purified by polysaccharide gel purification systems to obtain purified water-soluble Sparassis crispa polysaccharide (SCPS). The chemical structure of SCPS was analyzed by gas chromatography, Fourier transform infrared spectroscopy, methylation analysis, and nuclear magnetic resonance spectroscopy. The monosaccharide compositional analysis revealed that SCPS consisted of fucose, arabinose, galactose, glucose, xylose, mannose, ribose, galacturonic acid, glucuronic acid, and mannuronic acid in a molar ratio of 17.37:1.94:25.52:30.83:1.14:0.30:4.98:2.87:2.65. Moreover, the backbone of SCPS was composed of →3)-ß-d-Glcp-(1→4)-ß-d-Glcp-(1→, with side chains attached to the backbone at the O-6 positions through the →3,6)-ß-d-Glcp-(1→ linkage. The in vitro experiments were conducted to investigate the effects of SCPS on the proliferation and differentiation of mouse osteoblasts. The results showed that SCPS significantly enhanced the proliferation and differentiation of mouse osteoblasts, indicating their potential as a pharmaceutical agent for promoting osteoblast proliferation and differentiation.

First Report of Lecanicillium aphanocladii Causing Rot of Sparassis crispa in China.

Sparassis crispa, also known as cauliflower mushroom, is a new popularly edible mushroom in China, also a medicinal mushroom, which possesses various biological activities, such as immunopotentiation, anti-diabetes, anti-cancer, and anti-inflammatory effects. (Han et al., 2018). In recent years, the artificial cultivation of S. crispa has gained considerable public attention in China. In 2023, approximately 20% of S. crispa (about 0.05 ha of the planting area) showed obvious rot with white molds symptoms in mushroom hothouse, located in Shuangliu county, Sichuan province, China (GPS, 104°7'51"N, 30°25'2"E). Infected fruiting bodies were covered by white mycelia that later turned red or fuchsia. In the final stages of infection, the S. crispa fruiting bodies turned dark red or brown before rotting. The pathogen was isolated from the margin of the lesions by plating onto potato dextrose agar (PDA), and incubated at 25℃ in the dark for a week. Five pure culture fungal isolates were obtained. Collected isolates with similar morphology were described as Lecanicillium spp. (Zare et al., 2001). The colonies were raised, covered with white, the reverse side were violet brown, produced diffusing reddish-purple pigment. Conidiogenous cells produced singly, in pairs, verticillate or in dense irregular clusters on prostrate hyphae, at first flask-shaped, tapering into threadlike neck, with a size of 3.0-6.2×0.8-2.2 µm. Conidia were solitary, oval to subglobose, and 2.3-4.0×1.1-2.1 µm in size, similar to L. aphanocladii (Higo et al., 2021). For pathogenicity testing, ten fruiting bodies of S. crispa (planted in the bottles) were selected. Fungal cake of the isolate Bx-Ljb of L. aphanocladii were applied to the fruiting body of S. crispa, whereas pieces of sterile PDA medium were used as controls. All the bottles were incubated at 19±1℃, 85-100% relative humidity, and 18 h of light in the mushroom hothouse. A week later, the inoculated fruiting bodies developed brown spots and gradually expanding, with symptoms similar to the original diseased fruiting bodies. The controls remained healthy. The same fungus was reisolated from the infected fruiting bodies and subsequently identified by morphological characteristics and DNA sequence analysis. The pathogenicity test was repeated three times with similar results. For molecular identification, the DNA of the isolates was extracted using a Fungi Genomic DNA Extraction kit (Solarbio, Beijing). The SSU, LSU, and TEF1-α genes were amplified with the primer as previously described (Zhou et al., 2018). The generated sequences were deposited in GenBank with accession numbers OR206377, OR206378, and OR204702, respectively. BLASTn analyses showed >99.2% identity with previously deposited sequences of L. aphanocladii. Based on the maximum likelihood method, phylogenetic analysis revealed 99% bootstrap support values with L. aphanocladii. The fungus was identified as L. aphanocladii based on morphological and multilocus phylogenetic analyses. To our knowledge, there are two reports of L. aphanocladii on fruiting bodies of Tremella fuciformis and Morchella sextelata in China, and this is the first report of this fungus causing rot of S. crispa in China. It may be a reminder that the risk of L. aphanocladii in mushroom production in China is gradually increasing. These results will contribute to developing managemental strategies for this disease in S. crispa.

Neuroprotective Effects of Sparassis crispa Ethanol Extract through the AKT/NRF2 and ERK/CREB Pathway in Mouse Hippocampal Cells.

Sparassis crispa, known as the "Cauliflower mushroom", is an edible medicinal fungus found in Asia, Europe, and North America. Its fruiting bodies contain active biological and pharmacological ingredients with antitumor and anti-inflammatory properties. In this study, we investigated the neuroprotective effect of various Sparassis crispa extract against glutamate-induced toxicity and oxidative stress in hippocampal HT22 cells. Cell viability and reactive oxygen species (ROS) analyses served to evaluate the neuroprotective effects of Sparassis crispa ethanol extract (SCE) and their fractions partitioned with ethyl acetate (EtOAc; SCE-E) and water (SCE-W) in HT22 cells. SCE and SCE-E treatment reduced glutamate-induced cell death and ROS generation. SCE-E reduced apoptosis and ROS levels by regulating anti-apoptotic proteins. Under glutamate treatment, SCE-E activated nuclear factor erythroid-derived 2-related factor 2 (Nrf2) and regulated extracellular signal-regulated kinase (ERK) and AKT signals at late stages. SCE-E increased the protein expression of cAMP response element binding (CREB), brain-derived neurotrophic factor (BDNF), and Kelch-like ECH-associated protein 1 (Keap1), and decreased the Nrf2 protein expression. Moreover, co-treatment of SCE-E and wortmannin did not activate Nrf2 expression. Thus, the neuroprotective effect of SCE-E is likely due to Nrf2 and CREB activation through AKT and ERK phosphorylation, which effectively suppress glutamate-induced oxidative stress in HT22 cells. Accordingly, a daily supplement of SCE-E could become a potential treatment for oxidative-stress-related neurological diseases.

Neuroprotective effects of polysaccharide from Sparassis crispa on Alzheimer's disease-like mice: Involvement of microbiota-gut-brain axis.

Alzheimer's disease (AD) is an irreversible neurodegenerative disease that may cause neurotoxicity and imbalance in gut microbiota. A polysaccharide derived from Sparassis crispa-1 (SCP-1) acts as a neuroprotective agent in vitro. There is, however, no clarity on the mechanism responsible for SCP-1's neuroprotective effects against AD. In this study, C57BL/6J male mice were treated with D-galactose and AlCl3 to establish an animal model of AD, followed by treatment with SCP-1. As evidenced by behavioral tests and brain pathology, SCP-1 treatment ameliorated learning deficits and defective spatial recognition, reduced amyloidogenesis, and modulated the neurotransmitter levels (γ-aminobutyric acid, glutamate, and acetylcholine) in the brain of AD mice. The results of 16S rRNA sequencing revealed that SCP-1 reshaped the gut microbiota composition, especially by promoting the proliferation of butyrate-producing genera, such as Intestinaimonas, [Eubacterium] ventriosum group, Lachnospiraceae_UCG_010, and Lachnospiraceae_UCG_001, and suppressing the growth of inflammation-related bacteria (i.e., Escherichia/Shigella). Furthermore, SCP-1 significantly attenuated inflammation by reducing the levels of inflammatory cytokines, maintaining intestinal barrier function, inhibiting glial activation, and decreasing the expression of toll-like receptor 4 (TLR4) and nuclear factor-κB (NF-κB). Collectively, our findings suggest that SCP-1 may prevent the development of AD via modulation of gut microbiota and suppression of inflammation, for a potential application in preventing or managing AD.

Natural Products and Disease Prevention, Relief and Treatment.

This Special Issue focusses on the role of natural products in disease prevention, relief and treatment [...].

Comprehensive analysis of Sparassis crispa polysaccharide characteristics during the in vitro digestion and fermentation model.

Sparassis crispa (S.crispa), an edible mushroom, is widely used as a natural medicine due to its excellent pharmacological activities, including antitumor, anti-angiogenic, and immunomodulatory activities. In the current study, the digestion and fermentation characteristics of an S.crispa polysaccharide (SCP-1) were investigated by the in vitro simulated models. Our results revealed that SCP-1 was not degraded during the simulated gastrointestinal tract. However, it was consumed by human intestinal microbiota, which was characterized by enhancing the production of short-chain fatty acids (such as acetate, propionate, and butyrate), and modifying the gut microbiota composition through promoting beneficial genera (Prevotella 9, Dialister, Megamonas, and Megasphaera) and inhibiting proliferation of some harmful bacteria (i.e., Escherichia/Shigella). The PICRUSt prediction analysis indicated that SCP-1 significantly increased carbohydrate, energy, and amino acid metabolism. These results suggest that SCP-1 could be developed as a prebiotic addition and may improve host health by regulating gut microbiota.

Purification, structural characterization and neuroprotective effect of a neutral polysaccharide from Sparassis crispa.

In the present study, a purified polysaccharide (named SCP-1, Mw 1.368 × 104 Da) was isolated from Sparassis crispa, and its biological activity was evaluated in an oxidative stress model caused by H2O2 in hippocampal neuronal HT22 cells. SCP-1 was a heteropolysaccharide mainly comprising glucose, galactose, fucose, and mannose in a molar ratio of 52.10: 31.10: 15.04: 1.76. The main backbone of SCP-1 was predominantly composed of (1 â†’ 6)-α-D-Galp, (1 â†’ 6)-ß-D-Glcp, (1 â†’ 3)-ß-D-Glcp, (1 â†’ 2,6)-α-D-Galp and (1 â†’ 3,6)-ß-D-Glcp. The branches, substituted at the O-2 of Gal and O-3 of Glc, contained (1 â†’ 6)-2-OMe-α-D-Galp, (1 â†’ 4)-ß-D-Glcp, (1 â†’ 3)-ß-D-Glcp, and terminated by T-α-L-Fucp and T-ß-D-Glcp. Besides, SCP-1 could effectively protect the HT22 cells against H2O2-induced oxidative injury via decreasing the intracellular reactive oxygen species levels, modulating antioxidant enzymes, and reducing cell apoptosis. The findings suggested that SCP-1 holds a potential to be a natural antioxidant or as a neuroprotective agent.

Metabolomic analysis of antimicrobial mechanism of polysaccharides from Sparassis crispa based on HPLC-Q-TOF/MS.

Abuse of antibiotics makes antibiotic-resistance become a huge challenge in bacterial infection treatment. The discovery of new antibiotics is of great significance to human health. In this study, the antibacterial mechanism of Sparassis crispa polysaccharides (SCPs) was explored. The SCPs isolated from Sparassis crispa was composed of fucose, glucose and galactose with a molar ratio of 0.043 : 0.652: 0.305. Bacteriostatic tests showed SCPs inhibited the growth of Staphylococcus aureus better than Escherichia coli's, and damage to bacteria was observed under scanning electron microscopy. Metabolomic analysis based on HPLC-Q-TOF/MS indicated that SCPs disrupted metabolism of the glycolysis and tricarboxylic acid cycle pathways in S. aureus. The variations of fructose-1,6-diphosphate, 1,3-diphosphoglycerol, succinate and oxaloacetate were significant, whose systematic changes accompanied with decrease of ATP in cells indicated that SCPs could exert antibacterial effects by inducing dysfunction of catabolism and energy metabolism. Our research confirmed the antibacterial properties of SCPs and provided a perspective for understanding antibacterial mechanism of polysaccharides from natural products through metabolomics technology.

The complete mitochondrial genome of an edible mushroom, Sparassis crispa.

Sparassis crispa, also known as cauliflower mushroom, is a widely used medicinal mushroom in traditional Chinese medicine due to the presence of bioactive substances with pharmacological activity. Here, we report a complete mitochondrial genome sequence of S. crispa consisting of 139,253 bp containing 47 genes including 15 protein-coding genes, 27 transfer RNA, and 5 ribosomal RNA genes obtained from 40.406 Mb genome containing 18,917 predicted contigs using raw data of next-generation sequencing having 85.4% Q30. The overall base composition of S. crispa was 26.47% G-C and 73.53% A-T. The phylogenetic tree based on atp6 sequence data showed its close relationship with Sparassis radicata. The complete mitochondrial genome sequence of S. crispa provides an essential and important DNA molecular data for further phylogenetic and evolutionary analysis of S. crispa.

Mushroom Sparassis crispa (Hanabiratake) Fermented with Lactic Acid Bacteria Significantly Enhances Innate Immunity of Mice.

Sparassis crispa (SC; Japanese name: Hanabiratake) is a mushroom with high ß(1-3)-glucan content. We here studied the effects of SC and lactic acid bacteria-fermented SC (SCL) on innate immunity. In in vivo studies using mice, oral administration of SC or SCL enhanced the accumulation of macrophages, neutrophils, natural killer (NK) cells, and C-C chemokine receptor type 2- or phospho-Syk-expressing cells in the jejunum epithelial villi and spleen, with significantly higher cell numbers in the SCL group than in the SC group. In addition, mRNA levels of genes encoding tissue factor (TF) and tumor necrosis factor (TNF)-α were increased in monocytes/macrophages from the peritoneal cavity of mice orally administered SCL. In in vitro studies using cultured human monocytes, SC and SCL enhanced the expression of gees involved in blood coagulation and inflammation, as well as those encoding various innate immune-related factors, such as TF, TNF-α, plasminogen activator inhibitor (PAI)-1, monocyte chemotactic protein (MCP)-1, interleukin (IL)-1ß, IL-8, IL-12ß, and IL-17, in a dose-dependent manner. In particular, the expression levels of all these factors in monocytes were significantly higher with SCL treatment than with SC treatment. SCL significantly enhanced the phagocytosis of pH-sensitive fluorescent dye-labeled Escherichia coli by human monocytes compared to SC. The effect of SCL on phagocytosis was significantly reduced to approximately 30% by pre-digestion of SCL with ß-glucanase, suggesting that ß(1-3)-glucan in SCL is a major contributor to the effect. These data suggest that oral administration of SCL significantly enhances innate immunity in mice and possibly humans.

Nanocomposite Gel as Injectable Therapeutic Scaffold: Microstructural Aspects and Bioactive Properties.

The development of tissue scaffolds able to provide proper and accelerated regeneration of tissue is a main task of tissue engineering. We developed a nanocomposite gel that may be used as an injectable therapeutic scaffold. The nanocomposite gel is based on biocompatible gelling agents with embedded nanoparticles (iron oxide, silver, and hydroxyapatite) providing therapeutic properties. We have investigated the microstructure of the nanocomposite gel exposed to different substrates (porous materials and biological tissue). Here we show that the nanocomposite gel has the ability to self-reassemble mimicking the substrate morphology: exposition on porous mineral substrate caused reassembling of nanocomposite gel into 10× smaller scale structure; exposition to a section of humerus cortical bone decreased the microstructure scale more than twice (to ≤3 µm). The reassembling happens through a transitional layer which exists near the phase separation boundary. Our results impact the knowledge of gels explaining their abundance in biological organisms from the microstructural point of view. The results of our biological experiments showed that the nanocomposite gel may find diverse applications in the biomedical field.

Rapid Characterization of Chemical Components in Edible Mushroom Sparassis crispa by UPLC-Orbitrap MS Analysis and Potential Inhibitory Effects on Allergic Rhinitis.

Sparassis crispa is a kind of edible fungus widely grows in the north temperate zone, which shows various medicinal properties. Due to the complexity of chemical constitutes of this species, few investigations have acquired a comprehensive configuration for the chemical profile of it. In this study, a strategy based on ultra-high performance liquid chromatography (UPLC) combined with Orbitrap mass spectrometer (MS) was established for rapidly characterizing various chemical components in S. crispa. Through the summarized MS/MS fragmentation patterns of reference compounds and systematic identification strategy, a total of 110 components attributed to six categories were identified for the first time. Moreover, allergic rhinitis (AR) is a worldwide inflammatory disease seriously affecting human health, and the development of drugs to treat AR has been a topic of interest. It has been reported that the extracts of S. crispa showed obvious inhibitory effects on degranulation of mast cell- and allergen-induced IgE and proinflammatory mediators, but the active components and specific mechanism were still not clear. Src family kinases (SFKs) participate in the initial stage of allergy occurrence, which are considered the targets of AR treatment. Herein, on the basis of that self-built chemical database, virtual screening was applied to predict the potential SFKs inhibitors in S. crispa, using known crystal structures of Hck, Lyn, Fyn, and Syk as receptors, followed by the anti-inflammatory activity evaluation for screened hits by intracellular calcium mobilization assay. As results, sparoside A was directly confirmed to have strong anti-inflammatory activity with an IC50 value of 5.06 ± 0.60 µM. This study provides a useful elucidation for the chemical composition of S. crispa, and demonstrated its potential inhibitory effects on AR, which could promote the research and development of effective agents from natural resources.

Sparassis crispa exerts anti-inflammatory activity via suppression of TLR-mediated NF-κB and MAPK signaling pathways in LPS-induced RAW264.7 macrophage cells.

ETHNOPHARMACOLOGY RELEVANCE: Sparassis crispa, also known as cauliflower mushroom, has been used historically in traditional Asian medicine. It possesses various biological activities, such as immunopotentiation, anti-diabetes, anti-cancer, and anti-inflammatory effects. Recently, we isolated the non-aqueous fraction from methanol extract of S. crispa (SCF4) by using water-organic solvent mixtures and high-performance liquid chromatography (HPLC). In the present study, we identified the anti-inflammatory activity and action mechanism of SCF4 in lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophage cells. MATERIALS AND METHODS: The chloroform layer isolated from S. crispa methanol extract was separated into seven fractions using preparative HPLC. The fractions were then applied to NO assay to identify the fraction with the best anti-inflammatory activity. The inflammation inhibitory effect and underlying mechanism of SCF4 in LPS-stimulated RAW264.7 cells were assessed using WST-1 assay, enzyme-linked immunosorbent assay (ELISA), ROS assay, and Western blot analysis. RESULTS: SCF4 significantly suppressed LPS-induced production of pro-inflammatory mediators, such as nitric oxide (NO) and prostaglandin E2 (PGE2), and pro-inflammatory cytokines, including tumor necrosis factor (TNF)-α, interleukin (IL)- 6, and IL-1ß, without cytotoxicity. In addition, SCF4 downregulated not only the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), but also the activation of nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs) stimulated by LPS. SCF4 also blocked the nuclear translocation of NF-κB via reduction of inhibitor of κB alpha (IκBα) degradation. Furthermore, SCF4 inhibited the phosphorylation of transforming growth factor beta-activated kinase 1 (TAK1), an important upstream factor of NF-κB and MAPK signaling mediated through toll-like receptor (TLR). CONCLUSIONS: These findings demonstrate for the first time the correlation between the anti-inflammatory activity of SCF4 and TLR-mediated NF-κB and MAPK signaling pathways in LPS-stimulated RAW264.7 macrophage cells, suggesting that the non-aqueous extract of S. crispa could be applied as a promising natural product for the prevention and treatment of inflammatory diseases.

GC-MS based metabolomics study of fermented stipe of Sparassis crispa.

GC-MS coupled with multivariate statistical analysis was performed to understand metabolites difference between pileus and stipe of Sparassis crispa (cauliflower mushroom). Metabolic changes of S. crispa after fermentation by different microorganisms were also investigated. PCA score plot showed a clear separation between pileus and stipe of S. crispa regardless of fermentation. However, OPLS-DA score plot showed clear separation among fermented S. crispa samples according to microbial strain used, indicating that both pileus and stipe fermented with the same strain showed similar pattern of metabolites. Fructose, lactic acid, citric acid, malic acid, and phosphoric acid were metabolites that contributed to the discrimination of fermented S. crispa samples. Results of this study provide novel insights into intrinsic characteristics of stipe of S. crispa which is cheaper than pileus as ingredient for alternative functional food.

Dietary Sparassis crispa Reduces Body Fat Mass and Hepatic Lipid Levels by Enhancing Energy Expenditure and Suppressing Lipogenesis in Rats.

Accumulation of abdominal fat triggers metabolic syndrome, which is a cluster of metabolic abnormalities, such as dyslipidemia, glucose intolerance, insulin resistance or hyperinsulinemia, and hypertension, that leads to the development of diabetes and cardiovascular disease. Mushrooms have been used as a foodstuff and folk medicine worldwide. Among these mushrooms, Sparassis crispa (SC) is a relatively newly cultivated edible and medicinal mushroom, which has been reported to have anti-diabetic and anti-hypertensive properties. However, little is known about the anti-obesity and anti-hyperlipidemic properties of SC. In the present study, we investigated the effects of dietary SC on lipid metabolism and energy expenditure in Sprague-Dawley rats with diet-induced obesity and diabetes, and conducted respiratory gas analysis to determine how energy metabolism is altered by SC. After feeding periods of 3 and 7 weeks, dietary SC had significantly reduced hepatic triacylglycerol and cholesterol contents in a dose-dependent manner. These changes were attributable to suppression of fatty acid and cholesterol synthesis in the liver and increased insulin sensitivity in the body. In addition, after a feeding period of 6 weeks, dietary SC significantly increased energy expenditure through carbohydrate oxidation, reducing abdominal fat mass after 7 weeks. In conclusion, our results indicate that in addition to the previously reported anti-diabetic and anti-hypertensive activities, dietary SC exhibits anti-obesity and anti-hyperlipidemic activities that help protect against metabolic syndrome.

Simultaneous determination of the bioactive compounds from Sparassis crispa (Wulf.) by HPLC-DAD and their inhibitory effects on LPS-stimulated cytokine production in bone marrow-derived dendritic cell.

Sparassis crispa (Wulf.) belonging to the family of Sparassidaceae, has been widely used as an edible mushroom due to its unique flavor and functions to improve health. In this study, the compounds isolated from the extract of this mushroom were simultaneously quantified by the developed HPLC-DAD method and evaluated for the inhibitory activities on the production of the LPS-stimulated cytokines (IL-12p40, IL-6, and TNF-α) in bone marrow-derived dendritic cells (BMDCs). The contents of this compounds were 0.1928 ± 0.0118, 4.4137 ± 0.0240, 0.5237 ± 0.0005, 2.7303 ± 0.0206 mg/g for sparoside A (1), methyl 2,4-dihydroxy-3-methoxy-6-methylbenzoate (2), sparalide A (3), and 5'-deoxy-5'-methylthioadenosine (4), respectively, which demonstrated that they are the major constituents of this mushroom. Thus, our results were found that the sparoside A (1), 5-hydroxy-7-methoxyphthalide (6), 5-methoxy-7-hydroxyphthalide (7), nicotinamide (10), and adenosine (11) inhibit LPS-stimualted cytokine production, compound 6 is the most potent inhibitory activities on the production of IL-12p40, IL-6, and TNF-α with IC50 values of 0.19, 0.18, and 0.91 µM, respectively.

Effects of Sparassis crispa in Medical Therapeutics: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

In this study, we investigated the therapeutic potential and medical applications of Sparassis crispa (S. crispa) by conducting a systematic review of the existing literature and performing a meta-analysis. The original efficacy treatment of the mushroom extract is considered primarily and searched in electronic databases. A total of 623 articles were assessed, 33 randomized controlled experiments were included after the manual screening, and some papers, review articles, or editorials that did not contain data were excluded. A comparative standard means difference (SMD) and a funnel plot between control and S. crispa groups were used as parameters to demonstrate the beneficial effects of S. crispa for diabetes and cancer treatment, as well as anti-inflammatory, anti-fungal and antioxidant activities. The meta-analysis was carried out using Review Manager 5.1 software. Although for therapeutic diabetes there was heterogeneity in the subgroup analysis (I² = 91.9%), the overall results showed statistically significant SMDs in major symptoms that decreased serum insulin levels (SMD = 1.92, 95% CI (1.10, 2.75), I² = 0%), wound rates (SMD = 3.55 (2.56, 4.54), I² = 40%) and contributions to an increase in nutrient intake content (SMD = 0.32 (-0.15, 0.78), I² = 0%). Simultaneously, the study confirmed the utility of S. crispa treatment in terms of not only anti-cancer activity (reduction of tumor activity and survival of cancer cells I² = 42 and 34%, respectively) but also anti-inflammatory, anti-fungal and antioxidant activities (I² = 50, 44, and 10%, respectively). Our findings suggest that S. crispa extracts are useful for prevention and treatment of human diseases and might be the best candidates for future medicines.

New Aromatic Compounds from the Fruiting Body of Sparassis crispa (Wulf.) and Their Inhibitory Activities on Proprotein Convertase Subtilisin/Kexin Type 9 mRNA Expression.

Successive chromatography of EtOAc-soluble extracts of the fruiting body of Sparassis crispa (Wulf.) resulted in isolation of four new aromatic compounds, sparoside A (1) and sparalides A-C (3-5), two new naturally occurring compounds, 2 and 6, and eight known compounds, 7-14. The chemical structures were determined by interpretation of nuclear magnetic resonance and mass spectrometry spectroscopic data. Extract, solvent-soluble fractions of the extract, and all of the pure compounds isolated from the fractions were subjected to the mRNA expression assay for proprotein convertase subtilisin/kexin type 9 (PCSK9). Among them, sparoside A (1), hanabiratakelide A (8), adenosine (11), and 5α,6α-epoxy-(22E,24R)-ergosta-8(14),22-diene-3ß,7ß-diol (14) exhibited potent inhibitory activities on PCSK9 mRNA expression, with IC50 values of 20.07, 7.18, 18.46, and 8.23 µM, respectively (berberine, positive control, IC50 = 8.04 µM), suggesting that compounds 1, 8, 11, and 14 are suitable for use in supplements to the statins for hyperlipidemia treatments.

Mitochondria Related Pathway Is Essential for Polysaccharides Purified from Sparassis crispa Mediated Neuro-Protection against Glutamate-Induced Toxicity in Differentiated PC12 Cells.

The present study aims to explore the neuro-protective effects of purified Sparassis crispa polysaccharides against L-glutamic acid (L-Glu)-induced differentiated PC12 (DPC12) cell damages and its underlying mechanisms. The Sparassis crispa water extract was purified by a DEAE-52 cellulose anion exchange column and a Sepharose G-100 column. A fraction with a molecular weight of 75 kDa and a diameter of 88.9 nm, entitled SCWEA, was obtained. SCWEA was identified with a triple helix with (1→3)-linked Rha in the backbone, and (1→2) linkages and (1→6) linkages in the side bone. Our results indicated that the pre-treatment of DPC12 cells with SCWEA prior to L-Glu exposure effectively reversed the reduction on cell viability (by 3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay) and reduced L-Glu-induced apoptosis (by Hoechst staining). SCWEA decreased the accumulation of intracellular reactive oxygen species, blocked Ca(2+) influx and prevented depolarization of the mitochondrial membrane potential in DPC12 cells. Furthermore, SCWEA normalized expression of anti-apoptotic proteins in L-Glu-explored DPC12 cells. These results suggested that SCWEA protects against L-Glu-induced neuronal apoptosis in DPC12 cells and may be a promising candidate for treatment against neurodegenerative disease.