A Capsicum annuum L. seed extract exerts anti-neuroexcitotoxicity in HT22 hippocampal neurons.

The hippocampal memory deficit stands out as a primary symptom in neurodegenerative diseases, including Alzheimer's disease. While numerous therapeutic candidates have been proposed, they primarily serve to delay disease progression. Given the irreversible brain atrophy or injury associated with these conditions, current research efforts are concentrated on preventive medicine strategies. Herein, we investigated whether the extracts of Capsicum annuum L. seeds (CSE) and Capsicum annuum L. pulp (CPE) have preventive properties against glutamate-induced neuroexcitotoxicity (one of the main causes of Alzheimer's disease) in HT22 hippocampal neuronal cells. Pretreatment with CSE demonstrated significant anti-neuroexcitotoxic activity, whereas CPE did not exhibit such effects. Specifically, CSE pretreatment dose-dependently inhibited the elevation of excitotoxic elements (intracellular calcium influx and reactive oxygen species; ROS) and apoptotic elements (p53 and cleaved caspase-3). In addition, the glutamate-induced alterations of neuronal activity indicators (brain-derived neurotrophic factor; BDNF and cAMP response element-binding protein phosphorylation; CREB) were significantly attenuated by CSE treatment. We also found that luteolin is the main bioactive compound corresponding to the anti-neuroexcitotoxic effects of CSE. Our results strongly suggest that Capsicum annuum L. seeds (but not its pulp) could be candidates for neuro-protective resources especially under conditions of neuroexcitotoxicity. Its underlying mechanisms may involve the amelioration of ROS-mediated cell death and BDNF-related neuronal inactivity and luteolin would be an active compound.

Ten weeks of Capsicum annuum L. extract supplementation did not change adipose tissue-derived hormones, appetite, body composition, and muscle strength when combined with resistance training in healthy untrained men: A clinical trial study.

Capsiate (CAP) is a nonpungent capsaicin analog (Capsicum annuum L. extract) that has been studied as a potential antiobesity agent. However, the interaction between chronic CAP supplementation and resistance training is not clear. The purpose of this study was to examine the changes in adipose tissue-derived hormones, body composition, appetite, and muscle strength after 10 weeks of resistance training, combined with chronic CAP supplementation in healthy untrained men. We hypothesized that CAP could induce higher benefits when combined with resistance training after 10 weeks of intervention compared to resistance training alone. Twenty-four young men (age, 22.0 ± 2.9) were randomized to either capsiate supplementation (CAP = 12 mg/day) or placebo (PL), and both groups were assigned to resistance training. Body composition, leptin and adiponectin concentrations, subjective ratings of appetite, energy intake, and exercise performance were assessed at before and after 10 weeks of progressive resistance training. There was a significant increase in body mass (P < .001), fat-free mass (CAP: 58.0 ± 7.1 vs. post, 59.7 ± 7.1 kg; PL: pre, 58.4 ± 7.3 vs. post, 59.8 ± 7.1 kg; P < .001), resting metabolic rate (CAP: pre, 1782.9 ± 160.6 vs. post, 1796.3 ± 162.0 kcal; PL: pre, 1733.0 ± 148.9 vs. post, 1750.5 ± 149.8 kcal; P < .001), maximal strength at 45 leg press (P < .001) and bench press (P < .001) in both groups, but no significant (P > .05) supplementation by training period interaction nor fat mass was observed. For subjective ratings of appetite, energy intake, leptin, and adiponectin, no significant effect of supplementation by training period interaction was observed (P > .05). In conclusion, 10 weeks of resistance training increased total body weight, muscle mass, and maximum strength in healthy untrained men; however, CAP supplementation (12 mg, 7 days per week) failed to change adipose tissue-derived hormones, appetite, body composition and muscle strength in this population. Registered under Brazilian Registry of Clinical Trials (RBR-8cz9kfq).

Adjusting function of camphor on primary metabolism in Cinnamomum camphora stressed by high temperature.

Cinnamomum camphora has great economic value for its wide utilization in traditional medicine and furniture material, and releases lots of monoterpenes to tolerate high temperature. To uncover the adjusting function of monoterpenes on primary metabolism and promoting their utilization as anti-high temperature agents, the photosynthetic capacities, primary metabolite levels, cell ultrastructure and associated gene expression were surveyed in C. camphora when it was blocked monoterpene biosynthesis with fosmidomycin (Fos) and fumigated with camphor (a typical monoterpene in the plant) under high temperature (Fos+38 °C+camphor). Compared with the control (28 °C), high temperature at 38 °C decreased the starch content and starch grain size, and increased the fructose, glucose, sucrose and soluble sugar content. Meanwhile, high temperature also raised the lipid content, with the increase of lipid droplet size and numbers. These variations were further intensified in Fos+ 38 °C treatment. Compared with Fos+ 38 °C treatment, Fos+ 38 °C+camphor treatment improved the starch accumulation by promoting 4 gene expression in starch biosynthesis, and lowered the sugar content by suppressing 3 gene expression in pentose phosphate pathway and promoting 15 gene expression in glycolysis and tricarboxylic acid cycle. Meanwhile, Fos+ 38 °C+camphor treatment also lowered the lipid content, which may be caused by the down-regulation of 2 genes in fatty acid formation and up-regulation of 4 genes in fatty acid decomposition. Although Fos+ 38 °C+camphor treatment improved the photosynthetic capacities in contrast to Fos+ 38 °C treatment, it cannot explain the variations of these primary metabolite levels. Therefore, camphor should adjust related gene expression to maintain the primary metabolism in C. camphora tolerating high temperature.

Beneficial Effects of Fermentation of Red Chili Pepper Using Lactococcus lactis subs. Cremoris RPG-HL-0136 in High-Fat Diet-Induced Obese Mice.

Red chili pepper is a beneficial natural spicy food that has antiobesity and antitype II diabetes effects, but it is not conducive to in-depth research as a dietary strategy to treat obesity. This study aims to investigate the beneficial effects of red chili pepper, fermented with a novel Lactococcus lactis subs. cremoris RPG-HL-0136. LC-MS/MS analysis is conducted to detect the content of capsaicin and dihydrocapsaicin, and no significant difference is observed between the nonfermented red chili pepper (NFP) (W/W) and the prepared L. lactis subs. cremoris RPG-HL-0136-fermented chili mixture (LFP). After establishing a high-fat diet-induced obese type II diabetic mouse model, the effects on weight gain, weight loss of liver and testicular fat, total cholesterol, triglyceride, fasting glucose, insulin, and homeostatic model assessment for insulin resistance in LFP were evaluated to be better than those in NFP following 10 weeks of interventions. All animal experiments were approved by the Institutional Animal Care and Use Committee of Xinxiang medical university. NFP and LFP could increase the expression of transient receptor potential vanilloid subfamily 1, peroxisome proliferator-activated receptor-alpha and caspase-2 in the high-fat mice. Compared with unfermented red chili pepper, the fermented red chili pepper complex significantly reduced LPS, tumor necrosis factor-alpha, and interleukin-6 in serum (P < .05). Intake of LFP significantly increased the expression of claudin-1 and occludin in the colon of the high-fat mice (P < .05), and there was no damage to the stomach and colon. This study provides scientific evidence that red chili pepper, fermented with L. lactis subs. cremoris RPG-HL-0136, may be beneficial for future treatment of obesity and accompanying diabetes. (IACUC.No.XYLL-20200019).

Phytotoxicity of Chemical Compounds from Cinnamomum camphora Pruning Waste in Germination and Plant Cultivation.

Much previous research has indicated most composts of pruning waste are characterized by potential phytotoxicity, it is highly correlated with the chemical compounds of raw materials. Cinnamomum camphora, a common kind of pruning waste in Southeast Asia and East Asia, is characterized by intense bioactivities due to complex chemical components. This study investigated the potential phytotoxicity of C. camphora pruning waste in light of germination and higher plant growth. C. camphora extracted from leaves completely inhibited seed germination and still showed suppression of root elongation at an extremely low dosage. C. camphora extract also displayed significant inhibition of nutrient absorption in tomato seedlings, including moisture, available nutrients (N, P and K) and key microelements (Fe, Mn, Zn and S). The gene expression of aquaporins and transporters of nitrate and phosphate was significantly up-regulated in roots. This could be regarded as a positive response to C. camphora extract for enhancing nutrient absorption. Moreover, the severe damage to the plasma membrane in roots caused by C. camphora extract might seriously affect nutrient absorption. Camphor is the main component of the C. camphora extract that may induce the phytotoxicity of plasma membrane damage, resulting in the inhibition of nutrient absorption and low biomass accumulation. This study provided a new understanding of the ecotoxicological effects of C. camphora pruning waste, indicating that the harmless disposal of pruning waste requires much attention and exploration in the future.

Camphor Seed Kernel Oil Beneficial Effects the Gut Microbiota of the Non-Alcoholic Fatty Liver Disease Mice.

INTRODUCTION: Non-alcoholic fatty liver disease (NAFLD) has become one of the most common liver diseases globally and its negative impact has grad- ually attracted attention. METHOD: In order to explore whether camphor seed oil has a certain link effect on NAFLD, we identified its remodeling of intestinal flora and liver function index through the use of camphor seed kernel oil as food treatment (CCSKO). RESULTS: Our results showed that camphor seed oil significantly improved the liver index, TG content and histopathology of the NAFLD mice compared with other groups. Meanwhile, we found significant differences in the intestinal microbiota of mice in different treatment groups. DISCUSSION: The CCSKO treatment might significantly increase the abundance of Bacteroidetes, which were involved in many important metabolic activities in the human colon, including carbohydrate fermentation, utilization of nitrogenous substances, and biotransformation of bile acids and other steroids. CONCLUSIONS: These findings indicate a strong contribution in shaping the gut microbiota by treatment groups, shedding light on the impact of camphor tree seed kernel oil on gut microbiota.

De novo assembly and comparative transcriptome analysis: novel insights into terpenoid biosynthesis in Chamaemelum nobile L.

KEY MESSAGE: Analysis of terpenoids content, transcriptome from Chamaemelum nobile showed that the content of the terpenoids in the roots was the highest and key genes involved in the terpenoids synthesis pathway were identified. Chamaemelum nobile is a widely used herbaceous medicinal plant rich in volatile oils, mainly composed of terpenoids. It is widely used in food, cosmetics, medicine, and other fields. In this study, we analyzed the transcriptome and the content and chemical composition of the terpenoids in different organs of C. nobile. Gas chromatography-mass spectrometry analysis showed that the total content of the terpenoids among C. nobile organs was highest in the roots, followed by the flowers. Illumina HiSeq 2500 high-throughput sequencing technology was used to sequence the transcripts of roots, stems, leaves, and flowers of C. nobile. We obtained 139,757 unigenes using the Trinity software assembly. A total of 887 unigenes were annotated to secondary metabolism. In total, 55,711 differentially expressed genes were screened among different organs of C. nobile. We identified 16 candidate genes that may be involved in the terpenoid biosynthesis from C. nobile and analyzed their expression patterns using real-time PCR. Results showed that the expression pattern of these genes was tissue-specific and had significant differential expression levels in different organs of C. nobile. Among these genes, 13 were expressed in roots with the highest levels. Furthermore, the transcript levels of these 13 genes were positively correlated with the content of α-pinene, ß-phellandrene, 1,8-cineole, camphor, α-terpineol, carvacrol, (E,E)-farnesol and chamazulene, suggesting that these 13 genes may be involved in the regulation of the synthesis of the volatile terpenoids. These results laid the foundation for the subsequent improvement of C. nobile quality through genetic engineering.

Borneol Dehydrogenase from Pseudomonas sp. Strain TCU-HL1 Catalyzes the Oxidation of (+)-Borneol and Its Isomers to Camphor.

Most plant-produced monoterpenes can be degraded by soil microorganisms. Borneol is a plant terpene that is widely used in traditional Chinese medicine. Neither microbial borneol dehydrogenase (BDH) nor a microbial borneol degradation pathway has been reported previously. One borneol-degrading strain, Pseudomonas sp. strain TCU-HL1, was isolated by our group. Its genome was sequenced and annotated. The genome of TCU-HL1 consists of a 6.2-Mbp circular chromosome and one circular plasmid, pTHL1 (12.6 kbp). Our results suggest that borneol is first converted into camphor by BDH in TCU-HL1 and is further decomposed through a camphor degradation pathway. The recombinant BDH was produced in the form of inclusion bodies. The apparent Km values of refolded recombinant BDH for (+)-borneol and (-)-borneol were 0.20 ± 0.01 and 0.16 ± 0.01 mM, respectively, and the kcat values for (+)-borneol and (-)-borneol were 0.75 ± 0.01 and 0.53 ± 0.01 s-1, respectively. Two plant BDH genes have been reported previously. The kcat and kcat/Km values of lavender BDH are about 1,800-fold and 500-fold lower, respectively, than those of TCU-HL1 BDH. IMPORTANCE: The degradation of borneol in a soil microorganism through a camphor degradation pathway is reported in this study. We also report a microbial borneol dehydrogenase. The kcat and kcat/Km values of lavender BDH are about 1,800-fold and 500-fold lower, respectively, than those of TCU-HL1 BDH. The indigenous borneol- and camphor-degrading strain isolated, Pseudomonas sp. strain TCU-HL1, reminds us of the time 100 years ago when Taiwan was the major producer of natural camphor in the world.

Identification of 1,8-cineole, borneol, camphor, and thujone as anti-inflammatory compounds in a Salvia officinalis L. infusion using human gingival fibroblasts.

Drinking or gargling Salvia officinalis L. infusion (sage infusion) is thought to soothe a sore throat, tonsillitis, and inflamed, red gums, although structure-based scientific evidence for the key anti-inflammatory compounds in sage infusion is scarce. Human gingival fibroblasts (HGF-1) were treated with sage infusion (SI) or SI fractions containing either its volatile components and water (aqueous distillate, AD) or its dry matter (DM) for six hours. SI, AD, and DM reduced a mean phorbol-12-myristate-13-acetate/ionomycin (PMA/I)-stimulated release of the pro-inflammatory interleukins IL-6 and IL-8 by more than 50% (p < 0.05). Cellular uptake experiments and subsequent GC-MS analysis using stable-isotope-labeled internal standards revealed the presence of 1,8-cineole, borneol, camphor, and α-/ß-thujone in SI-treated cells; LC-MS analysis demonstrated the presence of rosmarinic acid. A significant, more than 50% mean inhibition of PMA/I-induced IL-6 and IL-8 release was demonstrated for the volatile compounds 1,8-cineole, borneol, camphor, and thujone, but not for the nonvolatile rosmarinic acid when applied in concentrations representative of sage infusion. Therefore, the volatile compounds were found to be more effective than rosmarinic acid. 1,8-Cineole, borneol, camphor, and α-/ß-thujone chiefly contribute to the anti-inflammatory activity of sage infusion in human gingival fibroblasts.

Molecular cloning and functional characterization of borneol dehydrogenase from the glandular trichomes of Lavandula x intermedia.

Several varieties of Lavandula x intermedia (lavandins) are cultivated for their essential oils (EOs) for use in cosmetic, hygiene and personal care products. These EOs are mainly constituted of monoterpenes including camphor, which contributes an off odor reducing the olfactory appeal of the oil. We have recently constructed a cDNA library from the glandular trichomes (the sites of EO synthesis) of L. x intermedia plants. Here, we describe the cloning of a borneol dehydrogenase cDNA (LiBDH) from this library. The 780 bp open reading frame of the cDNA encoded a 259 amino acid short chain alcohol dehydrogenase with a predicted molecular mass of ca. 27.5 kDa. The recombinant LiBDH was expressed in Escherichia coli, purified by Ni-NTA agarose affinity chromatography, and functionally characterized in vitro. The bacterially produced enzyme specifically converted borneol to camphor as the only product with K(m) and k(cat) values of 53 µM and 4.0 × 10(-4) s(-1), respectively. The LiBDH transcripts were specifically expressed in glandular trichomes of mature flowers indicating that like other Lavandula monoterpene synthases the expression of this gene is regulated in a tissue-specific manner. The cloning of LiBDH has far reaching implications in improving the quality of Lavandula EOs through metabolic engineering.

Seasonal influence on gene expression of monoterpene synthases in Salvia officinalis (Lamiaceae).

Garden sage (Salvia officinalis L., Lamiaceae) is one of the most important medicinal and aromatic plants and possesses antioxidant, antimicrobial, spasmolytic, astringent, antihidrotic and specific sensorial properties. The essential oil of the plant, formed mainly in very young leaves, is in part responsible for these activities. It is mainly composed of the monoterpenes 1,8-cineole, α- and ß-thujone and camphor synthesized by the 1,8-cineole synthase, the (+)-sabinene synthase and the (+)-bornyl diphosphate synthase, respectively, and is produced and stored in epidermal glands. In this study, the seasonal influence on the formation of the main monoterpenes in young, still expanding leaves of field-grown sage plants was studied in two cultivars at the level of mRNA expression, analyzed by qRT-PCR, and at the level of end-products, analyzed by gas chromatography. All monoterpene synthases and monoterpenes were significantly influenced by cultivar and season. 1,8-Cineole synthase and its end product 1,8-cineole remained constant until August and then decreased slightly. The thujones increased steadily during the vegetative period. The transcript level of their corresponding terpene synthase, however, showed its maximum in the middle of the vegetative period and declined afterwards. Camphor remained constant until August and then declined, exactly correlated with the mRNA level of the corresponding terpene synthase. In summary, terpene synthase mRNA expression and respective end product levels were concordant in the case of 1,8-cineole (r=0.51 and 0.67 for the two cultivars, respectively; p<0.05) and camphor (r=0.75 and 0.82; p<0.05) indicating basically transcriptional control, but discordant for α-/ß-thujone (r=-0.05 and 0.42; p=0.87 and 0.13, respectively).

Ex vivo skin absorption of terpenes from Vicks VapoRub ointment.

BACKGROUND: The pharmaceutical market offers a wide range of inhalant drug products applied on the skin that contain essential oils and/or their isolated compounds, i.e. terpenes. Because there are few data concerning the skin penetration of terpenes, especially from complex carriers, the goal of this study was to determine the ex vivo skin absorption kinetics of chosen terpenes, namely eucalyptol, menthol, camphor, alpha-pinene, and beta-pinene, from the product Vicks VapoRub. MATERIAL/METHODS: Human cadaver skin was placed in a flow-through diffusion chamber and the product was applied for 15, 30, and 60 min. After the application time the skin was separated into layers using a tape-stripping technique: three fractions of stratum corneum and epidermis with dermis, and terpenes amounts in the samples were determined by gas-chromatography. RESULTS: The investigated terpenes showed different absorption characteristics related to their physicochemical properties and did not permeate through the skin into the acceptor fluid. Eucalyptol had the largest total accumulation in the stratum corneum and in the epidermis with dermis, while alpha-pinene penetrated into the skin in the smallest amount. CONCLUSIONS: The short time in which saturation of the stratum corneum with the terpenes occurred and the high accumulation of most of the investigated terpenes in the skin layers proved that these compounds easily penetrate and permeate the stratum corneum and that in vivo they may easily penetrate into the blood circulation.

Biotransformation of (+)-camphor by cultured cells of Eucalyptus perriniana.

Seven new biotransformation products were isolated from a jar fermentor culture of Eucalyptus perriniana following administration of (+)-camphor. The main product was (1S,4R,6S)-6-hydroxybornan-2-one 6-O-beta-D-glucopyranoside. The minor components were (1S,4R,6R)-6-hydroxybornan-2-one 6-O-beta-D-glucopyranoside, (1R,4S,6S)-6-hydroxybornan-2-one 6-O-beta-D-glucopyranoside, (1R,4R,5R)-5-hydroxybornan-2-one 5-O-beta-D-glucopyranoside, (1R,3R,4S)-3-hydroxybornan-2-one 3-O-beta-D-glucopyranoside, (1R,4R,7R)-8-hydroxybornan-2-one 8-O-beta-D-glucopyranoside and 2-(4-oxo-2,2,3-trimethylcyclopentyl)-ethyl-beta-D-glucopyranoside. All products were mono-glucosides and the oxygen function was introduced before glucosylation. Reduction of the ketone group of camphor was not observed.