N-feruloylserotonin Research Articles

N-Feruloyl Serotonin Attenuates Neuronal Oxidative Stress and Apoptosis in Aβ25-35-Treated Human Neuroblastoma SH-SY5Y Cells.

Amyloid-beta (Aβ) aggregation and deposition have been identified as a critical feature in the pathology of Alzheimer's disease (AD), with a series of functional alterations including neuronal oxidative stress and apoptosis. N-feruloyl serotonin (FS) is a plant-derived component that exerts antioxidant activity. This study investigated the protective effects of FS on Aβ25-35-treated neuronal damage by regulation of oxidative stress and apoptosis in human neuroblastoma SH-SY5Y cells. The radical scavenging activities increased with the concentration of FS, exhibiting in vitro antioxidant activity. The Aβ25-35-treated SH-SY5Y cells exerted neuronal cell injury by decreased cell viability and elevated reactive oxygen species, but that was recovered by FS treatment. In addition, treatment of FS increased anti-apoptotic factor B-cell lymphoma protein 2 (Bcl-2) and decreased the pro-apoptotic factor Bcl-2-associated X protein. The FS attenuated Aβ-stimulated neuronal apoptosis by regulations of mitogen-activated protein kinase signaling pathways. Moreover, activated CREB-BDNF signaling was observed by the treatment of FS in Aβ25-35-induced SH-SY5Y cells. These results demonstrate that FS shows potential neuroprotective effects on Aβ25-35-induced neuronal damage by attenuation of oxidative stress and apoptosis, and suggest that FS may be considered a promising candidate for the treatment of AD.

HPLC-MS Methodology for R. carthamoides Extract Quality Evaluation: A Simultaneous Determination of Eight Bioactive Compounds

Classified as an adaptogen, Maral root (Rhaponticum carthamoides, Leuzea carthamoides) is a herb that has long been used in Siberian as well as Russian alternative medicine. With over 200 substances found, this plant is a great source of bioactive compounds which have significant beneficial effects on human health and physical enhancement. Simultaneous quantification of the eight most therapeutic and abundant substances, i.e., 20-hydroxyecdysone (20-HE), kaempferol, hesperetin, quercetin, chlorogenic acid, N-feruloyl serotonin, cynaropicrin, and tracheloside belonging to various groups, such as ecdysteroids, flavonoids, phenolics, sesquiterpenes, and lignans, was performed for the first time through validated HPLC-MS. The evaluated parameters for method validation showed excellent linearity with R2 higher than 0.996, stability under various environmental factors with % RSD ≤ 2%, and recovery between 97 and 103% for all the studied compounds. Other validation parameters including selectivity, sensitivity, and precision were found to be within the acceptance criteria. The results of the stability studies provide information on the best combination of conditions for sample handling and storage. Generally, for almost every compound, exposure to light and elevated temperature for 96 h led to degradation; nevertheless, the acidic environment was beneficial for most of them.

Safflower Seed Extract Attenuates the Development of Osteoarthritis by Blocking NF-κB Signaling.

Although safflower seed extract exhibits pharmacological activity against various diseases, the effects of its individual compounds on osteoarthritis (OA) have not been elucidated. Here, we evaluated the effects of these extracts and their single compounds on OA. N-(p-Coumaroyl) serotonin and N-feruloyl serotonin, main components of safflower seed extract, were isolated by high-performance liquid chromatography. Under in vitro OA mimic conditions, the expression of the matrix metalloproteinases (MMPs) MMP3/13 and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) ADAMTS5 were reduced in mouse chondrocytes treated with safflower seed extract. Furthermore, the oral administration of safflower seed extract attenuated cartilage destruction in a mouse OA model induced by destabilization of the medial meniscus. N-(p-Coumaroyl) serotonin and N-feruloyl serotonin, but not serotonin, reduced MMP3, MMP13, and ADAMTS5 expression in IL-1β-treated chondrocytes. Additionally, they significantly blocked the nuclear factor-κB (NF-κB) pathway by inhibiting IκB degradation and p65 phosphorylation. Our results suggest that safflower seed extract and its single compounds can attenuate cartilage destruction by suppressing MMP and ADMATS5 expression. The anti-arthritic effects are mediated by NF-κB signaling and involve the inhibition of IκB degradation and p65 phosphorylation. These results indicate that safflower seed extract may serve as a novel therapeutic agent against OA.

The Protective Role of Feruloylserotonin in LPS-Induced HaCaT Cells.

Epidermal inflammation is caused by various bacterial infectious diseases that impair the skin health. Feruloylserotonin (FS) belongs to the hydroxycinnamic acid amides of serotonin, which mainly exists in safflower seeds and has been proven to have anti-inflammatory and antioxidant activities. Human epidermis mainly comprises keratinocytes whose inflammation causes skin problems. This study investigated the protective effects of FS on the keratinocyte with lipopolysaccharides (LPS)-induced human HaCaT cells and elucidated its underlying mechanisms of action. The mechanism was investigated by analyzing cell viability, PGE2 levels, cell apoptosis, nuclear factor erythroid 2-related factor 2 (Nrf2) translocation, and TLR4/NF-κB pathway. The anti-inflammatory effects of FS were assessed by inhibiting the inflammation via down-regulating the TLR4/NF-κB pathway. Additionally, FS promoted Nrf2 translocation to the nucleus, indicating that FS showed anti-oxidative activities. Furthermore, the antioxidative and anti-inflammatory effects of FS were found to benefit each other, but were independent. Thus, FS can be used as a component to manage epidermal inflammation due to its anti-inflammatory and anti-oxidative properties.

Metabolomic Profiling of the Host Response of Tomato (Solanum lycopersicum) Following Infection by Ralstonia solanacearum.

Tomato (Solanum lycopersicum) is an important dietary source of bioactive phytochemicals and active breeding programs constantly produce new cultivars possessing superior and desirable traits. The phytopathogenic Ralstonia solanacearum, the causal agent of bacterial wilt, is a highly destructive bacterial disease with a high economic impact on tomato production. This study followed an untargeted metabolomic approach involving four tomato cultivars and aimed at the identification of secondary metabolites involved in plant defense after infection with R. solanacearum. Liquid chromatography coupled to mass spectrometry (LC-MS) in combination with multivariate data analysis and chemometric modelling were utilized for the identification of discriminant secondary metabolites. The total of 81 statistically selected features were annotated belonging to the metabolite classes of amino acids, organic acids, fatty acids, various derivatives of cinnamic acid and benzoic acids, flavonoids and steroidal glycoalkaloids. The results indicate that the phenylpropanoid pathway, represented by flavonoids and hydroxycinnamic acids, is of prime importance in the tomato defense response. The hydroxycinnamic acids esters of quinic acid, hexoses and glucaric acids were identified as signatory biomarkers, as well as the hydroxycinnamic acid amides to polyamines and tyramine. Interestingly, the rapid and differential accumulation of putrescine, dopamine, and tyramine derivatives, along with the presence of a newly documented metabolite, feruloyl serotonin, were documented in the infected plants. Metabolite concentration variability in the different cultivar tissues point to cultivar-specific variation in the speed and manner of resource redistribution between the host tissues. These metabolic phenotypes provide insights into the differential metabolic signatures underlying the defense metabolism of the four cultivars, defining their defensive capabilities to R. solanacearum.

SEROTONIN AND SEROTONIN DERIVATIVES: NEW APPLICATION FOR VARIOUS SKIN DISORDERS

Objectives In the human body, serotonin is well known as a neurotransmitter that regulates mood, appetite and sleep. However, more than 90% of the serotonin in a human body is generated in gastrointestinal, stored in platelets, circulating whole body exept brain, and contributes to various physiological function. Plants also produce serotonin, and this phyto-serotonin is known to be involved in plant growth, flowering, mature and senescence. The most difference between phyto-serotonin and human serotonin is phyto-serotonin turns to various serotonin derivatives to store in flowers and fruits during mature and senescence. Serotonin derivatives were dramatically increased against pathogen attack. There are 4 types of major serotonin derivates. Among these 4 compounds, Caffeoylserotonin (CaS), Feruloylserotonin (FS) and Coumaroylserotonin (CS) are major components which mainly found from some plant extracts. Almost all human diseases are accompanie/d by ROS generation, so we investigated the antioxidant ability of serotonin derivates to make sure that they have a potential as therapeutic reagent. Findings As a result, serotonin and serotonin derivates were found to have up to 16 fold higher antioxidant levels than Trolox which is the standard material of the antioxidant assay. In addition, CaS and 5-HT have been shown to inhibit protein damage and DNA damage either. Serotonin derivatives promote cell proliferation and migration and inhibit melanogensis and cell apoptosis. Interestingly, 5-HT regulated cell proliferation and cell migration only through ERK/AP-1/MMP9 pathway while additional Akt/NF-κB/MMP9 pathway was involved in effects of CaS. Conclusions These results suggest that CaS can enhance keratinocyte proliferation and migration. Overall, serotonin derivatives might have potential as reagents beneficial for wound closing, cell regeneration and anti-melanogenesis.

Feruloylserotonin inhibits hydrogen peroxide-induced melanogenesis and apoptosis in B16F10 and SK-Mel-2 melanoma cells

Feruloylserotonin (FS) is a major bioactive component of safflower seeds, with documented strong antibacterial, anti-inflammatory, and free radical scavenging activities. Reactive oxygen species (ROS) can strongly induce melanogenesis and cell apoptosis. The present study aimed to investigate the ability of FS in preventing hydrogen peroxide (H2O2)-induced melanogenesis and cell apoptosis. Melanogenesis and apoptotic cell death were induced by transient exposure to H2O2 in B16F10 and SK-Mel-2 melanoma cells. FS significantly inhibited melanogenesis and cell death in both cell lines. FS inhibited H2O2-induced melanin production by down-regulating CREB/MITF/TYR signaling via inhibited intracellular cAMP accumulation. Additionally, FS induced extracellular regulated kinase activation, which led to the degradation of MITF and consequently decreased TYR expression and melanin production in H2O2-stimulated cells. Furthermore, FS inhibited H2O2-induced apoptotic cell death by maintaining mitochondrial membrane potential. Therefore, FS might have potential use for cosmetic whitening and as a therapeutic agent for hyperpigmentation disorder.

Serotonins of safflower seeds play a key role in anti-inflammatory effect in lipopolysaccharide-stimulated RAW 264.7 macrophages

Safflower (Carthamus tinctorius) seeds are well- known traditional oriental medicines that have long been used for the remedies of blood stasis and bone formation in east Asia. In this study, ethyl acetate (EtOAc) was used for extraction of the main chemical compounds from C. tinctorius seeds. Four major compounds were identified, acacetin, cosmosiin, N-feruloyl serotonin and N-(p-coumaroyl) serotonin. Each compound was evaluated for its inhibitory activity against the inflammatory process of macrophages. All compounds significantly inhibited production of lipopolysaccharide (LPS)- stimulated nitric oxide (NO) and pro-inflammatory cytokines. The protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were dramatically decreased by serotonins in a dose-dependent manner in LPS-stimulated RAW 264.7 macrophages. These results suggest that serotonin derivatives from safflower seeds may reduce inflammation- related diseases.

Anti-hyperglycemic activity of polyphenols isolated from barnyard millet (Echinochloa utilis L.) and their role inhibiting α-glucosidase

The extracts and chemical compounds isolated from barnyard millet (Echinochloa utilis) grains were investigated as part of a search for naturally derived anti-hyperglycemic medicinal plants. Among the five different solvent extracts, the ethyl acetate extract showed the lowest IC50 value against Saccharomyces cerevisiae α-glucosidase (70.2 μg/mL). Chromatography yielded eight phenolic compounds that may have been responsible for this effect. Among them, N-p-coumaroyl serotonin (1, CS), feruloyl serotonin (2, FS), and luteolin (5) potently inhibited α-glucosidase with IC50 values of 1.3–17.8 μM compared with those of deoxynojirimycin (DNJ, IC50 = 2.5 ± 0.1 μM) and acarbose (IC50 = 255.1 ± 15.6 μM). Additionally, for the first time, we found that CS and FS were significantly inhibited mammalian rat intestinal sucrase (IC50 of 3.0 and 8.2 μM) and reduced glucose content (73 and 52 % at 0.1 mg/wells) in Caco-2 (human intestinal epithelial) cells. Furthermore, oral glucose tolerance test revealed improved glucose tolerance following treatment with the barnyard millet grains extract by retarding the postprandial rise in blood glucose in vivo. These results suggest that barnyard millet grain can be used as a natural functional medicine to prevent and alleviate type-2 diabetes.

Preparative Separation of N-Feruloyl Serotonin and N-(p-Coumaroyl) Serotonin from Safflower Seed Meal Using High-Speed Counter-Current Chromatography.

High-speed counter-current chromatography (HSCCC) was successfully applied for the preparative separation and purification of N-feruloyl serotonin (NF) and N-(p-coumaroyl) serotonin (NP) from safflower seed meal. After the measurement of partition coefficient of the two target compounds in the two-phase solvent systems, the HSCCC was performed well with a two-phase solvent system composed of CHCl3-methanol-0.1 M HCl at a volume ratio of 1 : 1 : 1, v/v. The upper phase was used as stationary phase and the lower phase was used as mobile phase. Under the optimized condition, 7.5 mg NF and 6.9 mg NP were separated from 40 mg crude sample with the purity of 98.8 and 97.3%, respectively. The structures of the isolated compounds were identified by (1)H NMR and (13)C NMR.

Anti-pigmentation effect of serotonin alkaloid isolated from Korean barnyard millet (Echinochola utilis)

Investigation of the melanogenesis inhibitory effect of barnyard millet grain extracts (BME) led to isolation of compounds against mushroom tyrosinase and B16 melanoma cells. Chromatography yielded five phenolic compounds, among which n-p-coumaroyl serotonin (1, CS) exhibited potent non-competitive inhibition (IC50 =8.8 μM) compared with that of kojic acid (IC50=14.6 μM) against mushroom tyrosinase. More importantly, CS decreased melanin content to eight times lower (IC50 =11.6 μM) in B16 melanocytes than that of feruloyl serotonin (2, FS, IC50 =86.5 μM) without cytotoxicity. CS significantly inhibited the protein expression of tyrosinase, tyrosinase-related protein-1 (TRP-1), TRP-2, and microphthalmia-associated transcription factor using immunoblotting. Furthermore, molecular docking simulations were also carried out to investigate the structure related to the inhibitory activities. Therefore, BME can be used as a natural source of depigmentation to prevent melanogenesis.

Biotransformation of serotonin derivatives by the larvae of common cutworm (Spodoptera litura)

Seven serotonin derivatives (1–7) were biotransformed using larvae of Spodoptera litura. N-p-methoxy cinnamoyl serotonin (3) was converted into its O-demethylated compound, N-p-coumaroyl serotonin (2). Furthermore, N-feruloyl serotonin (5) and N-isoferuloyl serotonin (6) were converted into, two new compounds, N-feruloyl serotonin-4′-O-β-D-glucopyranoside (8) and N-isoferuloyl serotonin-3′-O-β-D-glucopyranoside (9), respectively. These structures were established by IR, 1D-NMR and 2D-NMR spectral studies.

Potent α‐Glucosidase Inhibitors from Safflower (Carthamus tinctorius L.) Seed

As part of the search for naturally derived α-glucosidase inhibitors, the chemical components isolated from safflower seed (Carthamus tinctorius L.) were evaluated. The compounds active as α-glucosidase inhibitors were serotonin derivatives (e.g. N-p-coumaroyl serotonin (1) and N-feruloyl serotonin (2)). These compounds showed a potent inhibitory activity, the 50% inhibitory concentration (IC(50) ) values were calculated as 47.2 µm (1) and 99.8 µm (2) while that of the reference drugs acarbose and 1-deoxynojirimycin were evaluated as 907.5 µm and 278.0 µm, respectively. Regarding the structure of the serotonin derivative, the existence of the hydroxyl group at 5-position in the serotonin moiety and the linkage of cinnamic acid and serotonin are essential for α-glucosidase inhibitory activities. These results are helpful for the proper use of safflower seed as traditional medicine for the treatment of diabetes, moreover, it could serve to develop medicinal preparations as supplements and functional foods for diabetes. In particular, the serotonin compounds could be used as a lead compound for a new potential α-glucosidase inhibitor derived from the plant.

Biosynthesis and biotechnological production of serotonin derivatives

Serotonin derivatives belong to a class of phenylpropanoid amides found at low levels in a wide range of plant species. Representative serotonin derivatives include feruloylserotonin (FS) and 4-coumaroylserotonin (CS). Since the first identification of serotonin derivatives in safflower seeds, their occurrence, biological significance, and pharmacological properties have been reported. Recently, serotonin N-hydroxycinnamoyl transferase (SHT), which is responsible for the synthesis of serotonin derivatives, was cloned from pepper (Capsicum annuum) and characterized in terms of its enzyme kinetics. Using the SHT gene, many attempts have been made to either increase the level of serotonin derivatives in transgenic plants or produce serotonin derivatives de novo in microbes by dual expression of key genes such as SHT and 4-coumarate-CoA ligase (4CL). Due to the strong antioxidant activity and other therapeutic properties of serotonin derivatives, these compounds may have high potential in treatment and prophylaxis, as cosmetic ingredients, and as major components of functional foods or feeds that have health-improving effects. This review examines the biosynthesis of serotonin derivatives, corresponding enzymes, heterologous production in plants or microbes, and their applications.

Ectopic expression of serotonin N-hydroxycinnamoyltransferase and differential production of phenylpropanoid amides in transgenic tomato tissues

Tomato contains high levels of amines such as serotonin and tyramine and is a suitable host to enhance phenylpropanoid amides (PAs), an important class of nutraceuticals with strong antioxidant activity and chemotherapeutic effects, by ectopic expression of the corresponding gene, serotonin N-hydroxycinnamoyltransferase (SHT). To assess whether ectopic overexpression of SHT cDNA under the control of the CaMV 35S promoter would enhance levels of PAs, we generated transgenic tomato plants and analyzed the levels of PAs. Transgenic tomato plants exhibited increased synthesis of PAs such as feruloylserotonin (FS), 4-coumaroylserotonin (CS), feruloyltyramine (FT), 4-coumaroyltyramine (CT), and feruloyloctopamine (FO) in 1-month-old leaves compared to the wild type. The increase and relative levels of PAs were even more apparent in 3-month-old leaves of transgenic tomato. When tomato leaves were challenged by wounding, levels of PAs in the best transgenic line increased by 3- and 10-fold for CS + FS and CT + FT, respectively. In contrast to leaves, tomato fruit only showed enhanced synthesis of CT + FT, whereas CS + FS levels were not enhanced. Regarding amine content, the levels of tyramine were much higher than those of serotonin in tomato leaves and fruits. The high levels of tyramine may contribute to the preferential production of CT + FT rather than CS + FS, although SHT enzyme shows the highest substrate affinity toward serotonin rather than tyramine.

Expression of serotonin derivative synthetic genes on a single self-processing polypeptide and the production of serotonin derivatives in microbes

The plant-specific serotonin derivatives feruloylserotonin (FS) and 4-coumaroylserotonin (CS) are synthesized by the enzymes 4-coumarate:coenzyme A ligase (4CL) and serotonin N-hydroxycinnamoyltransferase (SHT). To express these genes coordinately, SHT was fused in-frame with the self-processing FDMV 2A sequence followed by 4CL in a single open reading frame and introduced into Escherichia coli or Saccharomyces cerevisiae. The transgenes were abundantly expressed in both recombinant microbes, but functional expression was achieved only in yeast, with cleavage at the 2A sequence yielding monomeric SHT-2A and 4CL as judged by immunoblot and product analyses. In the presence of an exogenous supply of precursors such as serotonin and ferulic acid, the recombinant yeast synthesized 4.5 mg l(-1) FS in the medium while 0.02 mg l(-1) FS was produced in the cells. Time-course analysis indicated peak accumulation of FS at 24 h after induction, and this level was maintained until 96 h. The optimum precursor concentration was 2 mM. A series of serotonin derivatives was produced by adding various cinnamate derivative precursors with serotonin; 2.5 mg l(-1) caffeoylserotonin (CaS) and 1.4 mg l(-1) CS were produced, whereas no sinapoylserotonin or cinnamoylserotonin was yielded.

HPLC Analysis of Serotonin, Tryptamine, Tyramine, and the Hydroxycinnamic Acid Amides of Serotonin and Tyramine in Food Vegetables

Biogenic monoamines such as serotonin, tryptamine, and tyramine function as neurotransmitters and mitogenic factors in animals and are involved in flowering, morphogenesis, and protection from and adaptation to environmental changes in plants. In plants, serotonin and tyramine are conjugated to form phenolic compounds via thioester linkages during the synthesis of hydroxycinnamic acid amides, including p-coumaroylserotonin (CS), feruloylserotonin (FS), p-coumaroyltyramine (CT), and feruloyltyramine (FT). In this study, we determined the amounts of the biogenic monoamines CS, FS, CT, and FT in commonly consumed vegetables using high-performance liquid chromatography. Serotonin, tryptamine, and tyramine were detected in all vegetables tested. The serotonin levels ranged from 1.8 to 294 microg/g of dry weight, the tryptamine levels ranged from 0.8 to 372 microg/g of dry weight, and the tyramine levels ranged from 1.4 to 286 microg/g of dry weight. The highest serotonin and tryptamine contents were found in tomato and cherry tomato (140.3-222 microg/g of dry weight), while paprika and green pepper had higher tyramine contents than the other vegetables (286 and 141.5 microg/g of dry weight, respectively). Overall, the levels of CS, FS, CT, and FT ranged from 0.03 to 13.8 microg/g of dry weight, with green onion possessing the highest levels of CS (0.69 microg/g of dry weight), FT (1.99 microg/g of dry weight), and CT (13.85 microg/g of dry weight).

Endosperm-Specific Expression of Serotonin N-Hydroxycinnamoyltransferase in Rice

Serotonin N-hydroxycinnamoyltransferase (SHT) is a key enzyme in the synthesis of feruloylserotonin (FS) and 4-coumaroylserotonin (CS). These serotonin derivatives show strong antioxidant activity, making them valuable for both nutritional and pharmacological use in humans. Ectopic expression of SHT under the control of the endosperm specific-glutelin and prolamin promoters from rice was produced via Agrobacterium-mediated transformation. SHT expression was confirmed by Southern blot analysis, followed by Northern blotting and SHT enzyme activity analyses using total RNA and protein, respectively, extracted from transgenic seeds. The glutelin A3 (GluA3) promoter produced low SHT mRNA expression in rice seeds, whereas the prolamin promoter expressed high levels of SHT mRNA. In spite of the ectopic expression of SHT in rice seeds, both transgenic genotypes accumulated levels of serotonin derivatives similar to those found in wild-type rice. Furthermore, our data suggest that serotonin, rather than phenylpropanid-CoAs, is the rate-limiting substrate in the biosynthesis of serotonin derivatives in SHT-overexpressing transgenic rice seeds.