Optimized extraction, enrichment, identification and hypoglycemic effects of triterpenoid acids from Hippophae rhamnoides L pomace.

The Hippophae rhamnoides L. pomace was generated in the production process for juice, wine of food industry. To expand the application of pomace, the extraction process optimization, enrichment and identification of triterpene acids were performed in this study. The extraction yield was 14.87% under optimal ultrasound-assisted extraction techniques performed via response surface methodology. The extract was subsequently purified to obtain the triterpenoid acid enrichment fraction (TPF) with the content of 75.23% ± 1.45%. 13 triterpenoid acids were identified via UPLC-Triple-TOF MS/MS and further semi-quantified through comparison with triterpenoid acid standards. TPF exhibited a strong inhibitory effect on α-glucosidase with IC50 value of 5.027 ± 0.375 µg/mL, as determined via enzyme inhibition experiment and molecular docking. Additionally, the TPF significantly reduced postprandial glucose levels, as revealed via carbohydrate tolerance tests, as well as ameliorate serum lipid profiles. Therefore, pomace may be a promising resource of functional food components with therapeutic and commercial values.

Flavonoid Compounds in Hippophae rhamnoides L. Protect Endothelial Cells from Oxidative Damage Through the PI3K/AKT-eNOS Pathway.

Sea buckthorn, a traditional medicinal plant, has been used for several years in China for the prevention and treatment of various diseases, a practice closely associated with its significant antioxidant activity. The aim of this study was to investigate the protective effects of sea buckthorn flavonoids on vascular endothelial cells in an oxidative stress environment. We isolated and extracted active compounds from sea buckthorn and investigated their impact on endothelial nitric oxide synthase (eNOS) activity through the PI3K/AKT-eNOS signaling pathway through a combination of network pharmacology and cellular experiments, elucidating the regulatory effects of these compounds on endothelial cell functions. Three flavonoids, named Fr.4-2-1, Fr.4-2-2 and Fr.4-2-3, were obtained from sea buckthorn. The results of network pharmacology indicated that they might exert their effects by regulating the PI3K-AKT signaling pathway. In vitro results showed that all three flavonoids were effective in alleviating the degree of oxidative stress in cells, among which Fr.4-2-1 exerted its antioxidant effects by modulating the PI3K/AKT-eNOS pathway. Flavonoids in sea buckthorn can effectively inhibit oxidative stress-induced cellular damage, preserving the integrity and functionality of endothelial cells, which is crucial for maintaining vascular health and function.

Phenolic composition and bioactivities of sea buckthorn (Hippophae rhamnoides L.) fruit and seeds: an unconventional source of natural antioxidants in North America.

BACKGROUND: Sea buckthorn (Hippophae rhamnoides L.) was introduced into Canada in the early 2000s. This plant bears fruits with high commercial value in other countries due to its premium oil. Nevertheless, sea buckthorn berries are also a rich source of bioactives with nutraceutical potential, especially the variety grown in Newfoundland (Canada), which has not previously been characterized. As such, this study evaluated the composition of polyphenols in sea buckthorn pomace and seeds, as well as their prospective health-promoting effects. RESULTS: Polyphenolic identification by high-performance liquid chromatography-ultraviolet-mass spectrometry-time of flight revealed the presence of 24 compounds in the seeds and 16 compounds in the pomace, including phenolic acids, flavonoids, and tannins, with ellagic acid derivative IV (pomace, 52.13 µg g-1) and (+)-catechin (seeds, 690.8 µg g-1) being the most dominant. Sea buckthorn extracts displayed in vitro antidiabetic and anti-obesity potential by inhibiting α-glucosidase (71.52-99.31%) and pancreatic lipase (15.80-35.61%) enzymes, respectively. The extracts also protected low-density-lipoprotein cholesterol (50.97-89.67%) and supercoiled DNA (35.11-79.84%) from oxidative damage. CONCLUSION: Sea buckthorn berries grown in Canada showed promising health benefits induced by their rich and diverse polyphenolic profile and need to be considered for further commercial expansion as a bioactive-loaded superfruit. © 2024 Society of Chemical Industry.

Comparative quantitative analysis of fruit oil from Hippophae rhamnoides (seabuckthorn) by qNMR, FTIR and GC-MS.

Objective: To develop a qNMR method for quantitative analysis of triacylglycerols in fruit oil of Hippophae rhamnoides (seabuckthorn, SBT) and analyze commercial samples of SBT oils using GC-MS and FTIR. Methods: SBT fruit oil (IPHRFH) was extracted with hexane and the triglyceride (TAG) was isolated by vacuum liquid chromatography. Six different branded SBT oils purchased from e-commerce suppliers (Amazon) and in-house prepared SBT oil was analyzed by qNMR and fatty acyl composition of TAGs determined by using NMR. In-house oil was also analysed by GC-MS and FTIR spectroscopy. Results: The qNMR results showed that the oil contained 80.3% of triacylglycerol (TAG). The SBT oil TAGs comprised of linolenate 6.6%, palmitoleate/oleate 65.4%, and total saturated fatty acyl chain including palmitate 28% as determined by qNMR. GC-MS analysis revealed that the major acyl functionalities present in the TAG were palmitoleic acid 36.5%, oleic acid 12.9%, palmitic acid 21.2%, and linoleic acid 18%. Of the six commercial samples analyzed, samples from only one supplier (SW) were fruit oil; All others were the seed oils or mix of fruit oil and seed oil. The labels for samples except for the SW did not indicate whether it was fruit oil or seed oil. Conclusion: The results suggest that SBT oil should be analyzed by combination of GC-MS, FTIR and qNMR for factual content of free fatty acid or TAGs, which are chemically different in nature and affect the quality of oil. GC-MS showed the content of omega free fatty acids after hydrolysis, while qNMR and FTIR showed the content of TAGs. The major acyl functionalities found in SBT fruit oil TAGs are palmitoleate/palmitate/oleate, while linoleate and linonelate make up a minor fraction. Furthermore, analysis of commercial samples showed discrepancies between label claims and actual content.

In Search of Authenticity Biomarkers in Food Supplements Containing Sea Buckthorn: A Metabolomics Approach.

Sea buckthorn (Hippophae rhamnoides L.) (SB) is increasingly consumed worldwide as a food and food supplement. The remarkable richness in biologically active phytochemicals (polyphenols, carotenoids, sterols, vitamins) is responsible for its purported nutritional and health-promoting effects. Despite the considerable interest and high market demand for SB-based supplements, a limited number of studies report on the authentication of such commercially available products. Herein, untargeted metabolomics based on ultra-high-performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry (UHPLC-QTOF-ESI+MS) were able to compare the phytochemical fingerprint of leaves, berries, and various categories of SB-berry herbal supplements (teas, capsules, tablets, liquids). By untargeted metabolomics, a multivariate discrimination analysis and a univariate approach (t-test and ANOVA) showed some putative authentication biomarkers for berries, e.g., xylitol, violaxanthin, tryptophan, quinic acid, quercetin-3-rutinoside. Significant dominant molecules were found for leaves: luteolin-5-glucoside, arginine, isorhamnetin 3-rutinoside, serotonin, and tocopherol. The univariate analysis showed discriminations between the different classes of food supplements using similar algorithms. Finally, eight molecules were selected and considered significant putative authentication biomarkers. Further studies will be focused on quantitative evaluation.

Preparation of Cosmetic Emulsions Containing Hippophae Oil Isolated by Various Methods: Study of Their Antioxidant, Sun-Blocking and Physicochemical Properties.

An industry listed as one of the largest globally is the cosmetic industry. In recent years, this industry has shown growing interest in the application of natural ingredients providing advanced properties to cosmetic creams such as moisturizing, antioxidant, sun-protecting and antimicrobial effects. In this context, the present study concerns the production of cosmetic emulsions containing hippophae oil obtained via the methods of extraction, hydro-distillation and maceration using sunflower oil as the carrier oil. Firstly, an IR-ATR analysis was performed showing that the oils prepared were close to those commercially obtained. Then, the stability of the emulsions was tested over a time period of four months through measuring their pH and viscosity values with positive outcomes, and their antioxidant ability was also measured using the DPPH method. The latter one showed that hippophae oil greatly improves the antioxidant capacity. Moreover, based on the fact that sea buckthorn contains carotenoids, the SPF value of the emulsions was determined. The results showed that the addition of hippophae oil to the emulsions gave higher absorption in UV-Vis, thus higher SPF values. Py-GC/MS analysis was used to identify decomposition compounds in the produced oils. Among those, valuable compounds such as Ω-6, Ω-7 and Ω-9 fatty acids and many aldehydes were found by the decomposition of the oils.

Optimization of Enzyme-Assisted Extraction of Bioactive Compounds from Sea Buckthorn (Hippophae rhamnoides L.) Leaves: Evaluation of Mixed-Culture Fermentation.

Hippophae rhamnoides L. leaves possess a remarkable amount of polyphenols that could serve as a natural remedy in various applications. In comparison, numerous techniques, such as conventional and high-pressure techniques, are available for extracting the bioactive fractions from sea buckthorn leaves (SBL). However, enzyme-assisted extraction (EAE) of SBL has not been comprehensively studied. The aim of this study was to optimize critical EAE parameters of SBL using the cellulolytic enzyme complex, Viscozyme L, to obtain a high-yield extract with a high concentration of bioactive compounds. In order to determine the optimal conditions for EAE, the study employed a central composite design and response surface methodology to analyze the effects of four independent factors (pH, temperature, extraction time, and enzyme concentration) on two different responses. Our findings indicated that under optimal conditions (3:15 h extraction, temperature 45 °C, pH 4.9, and 1% Viscozyme L v/w of leaves DW), EAE yielded 28.90 g/100 g DW of the water-soluble fraction. Furthermore, the EAE-optimized liquid extract was continuously fermented using an ancient fermentation starter, Tibetan kefir grains, which possess lactic acid bacteria (LAB) and have significant potential for use in biopreservation. Interestingly, the results indicated various potential prebiotic characteristics of LAB. Additionally, alterations in the cell wall morphology of the SBL residue after EAE were examined using scanning electron microscopy (SEM). This study significantly optimized EAE parameters for sea buckthorn leaves, providing a promising natural source of bioactive compounds for various applications, such as nutraceuticals, functional foods, and high-value products.

HrTPS12 gene dramatically enhanced insect resistance of sea buckthorn to infection by fruit fly (Rhagoletis batava obseuriosa Kol.).

BACKGROUND: Terpenoids emitted from plants are important for regulating plant-insect interaction. However, it is still unclear how terpenoids affect the host defense system. There are few reports of terpenoids' involvement in the mechanisms that regulate woody plants' insect resistance. RESULTS: The (E)-ß-ocimene of terpenes was only found in RBO-resist leaves, and its content was higher than that of other type terpenes. Further, we also found (E)-ß-ocimene had a significant avoidance effect on RBO and reached 87.5% of the highest avoidance rate. Meanwhile, overexpression of HrTPS12 in Arabidopsis increased the HrTPS12 expression level, (E)-ß-ocimene content, and enhanced the defense against RBO. However, silencing HrTPS12 in sea buckthorn revealed that the expression levels of HrTPS12 and (E)-ß-ocimene significantly decreased, causing the attraction effect on RBO. CONCLUSION: HrTPS12 was an up-regulator, which improves sea buckthorn resistance to RBO by regulating the synthesis of volatile (E)-ß-ocimene. These results provide in-depth information about the interaction between RBO and sea buckthorn and provide a theoretical basis for developing plant-based insect repellents that can be used to manage RBO. © 2023 Society of Chemical Industry.

Flavonoids from Seabuckthorn (Hippophae rhamnoides L.) mimic neurotrophic functions in inducing neurite outgrowth in cultured neurons: Signaling via PI3K/Akt and ERK pathways.

BACKGROUND: Various brain disorders, including neurodegenerative diseases and major depressive disorders, threaten an increasing number of patients. Seabuckthorn, a fruit from Hippophae rhamnoides L., is an example of "medicine food homology". The fruit has enriched flavonoids that reported to have benefits in treating cognitive disorders. However, the studies on potential functions of Seabuckthorn and/or its flavonoid-enriched fraction in treating neurodegenerative disorders are limited. PURPOSE: This study aimed to determine the ability and mechanism of the flavonoid-enriched fraction of Seabuckthorn (named as SBF) in mimicking the neurotrophic functions in inducing neurite outgrowth of cultured neurons. METHODS: Cultured PC12 cell line, SH-SY5Y cell line and primary neurons (cortical and hippocampal neurons isolated from E17-19 SD rat embryos) were the employed models to evaluate SBF in inducing neurite outgrowth by comparing to the effects of NGF and BDNF. Immuno-fluorescence staining was applied to identify the morphological change during the neuronal differentiation. Luciferase assay was utilized for analyzing the transcriptional regulation of neurofilaments and cAMP/CREB-mediated gene. Western blot assay was conducted to demonstrate the expressions of neurofilaments and phosphorylated proteins. RESULTS: The application of SBF induced neuronal cell differentiation, and this differentiating activation was blocked by the inhibitors of PI3K/Akt and ERK pathways. Additionally, SBF showed synergy with neurotrophic factors in stimulating the neurite outgrowth of cultured neurons. Moreover, the major flavonoids within SBF, i.e., isorhamnetin, quercetin and kaempferol, could account for the neurotrophic activities of SBF. CONCLUSION: Seabuckthorn flavonoids mimicked neurotrophic functions in inducing neuronal cell differentiation via activating PI3K/Akt and ERK pathways. The results suggest the beneficial functions of Seabuckthorn as a potential health food supplement in treating various brain disorders, e.g., neurodegenerative diseases.

Hippophae rhamnoides L.: A Comprehensive Review on the Botany, Traditional Uses, Phytonutrients, Health Benefits, Quality Markers, and Applications.

Hippophae rhamnoides L. (sea buckthorn), consumed as a food and health supplement worldwide, has rich nutritional and medicinal properties. Different parts of H. rhamnoides L. were used in traditional Chinese medicines for relieving cough, aiding digestion, invigorating blood circulation, and alleviating pain since ancient times. Phytochemical studies revealed a wide variety of phytonutrients, including nutritional components (proteins, minerals, vitamins, etc.) and functional components like flavonoids (1-99), lignans (100-143), volatile oils (144-207), tannins (208-230), terpenoids (231-260), steroids (261-270), organic acids (271-297), and alkaloids (298-305). The pharmacological studies revealed that some crude extracts or compounds of H. rhamnoides L. demonstrated various health benefits, such as anti-inflammatory, antioxidant, hepatoprotective, anticardiovascular disease, anticancer, hypoglycemic, hypolipidemic, neuroprotective, antibacterial activities, and their effective doses and experimental models were summarized and analyzed in this paper. The quality markers (Q-markers) of H. rhamnoides L. were predicted and analyzed based on protobotanical phylogeny, traditional medicinal properties, expanded efficacy, pharmacokinetics and metabolism, and component testability. The applications of H. rhamnoides L. in juice, wine, oil, ferment, and yogurt were also summarized and future prospects were examined in this review. However, the mechanism and structure-activity relationship of some active compounds are not clear, and quality control and potential toxicity are worth further study in the future.

Development of fermented sea buckthorn (Hippophae rhamnoides L.) juice and investigation of its antioxidant and antimicrobial activity.

Sea buckthorn (Hippophae rhamnoides L.) is an edible and medicinal plant species. However, due to its sour taste, it is not readily accepted by consumers. To overcome this, fermentation can be used to change its flavor profile. In this study, we used response surface methodology (RSM) to determine the best process for producing fermented sea buckthorn juice (FSBJ) using probiotics. The biological enzyme activity and total flavonoid content (TFC) of sea buckthorn juice (SBJ) increased after fermentation. When the number of bacteria inoculated was 4.08 × 106 CFU/mL and the inoculation ratio was 30% Z. mobilis, 5% L. casei, 13.75% L. plantarum, 31.25% P. acidilactici, 12.5% L. animalis, and 7.5% P. pentosaceus, the amount of sugar was 2.98% (w/v) after 20 h of fermentation at 37°C, and the superoxide dismutase (SOD) activity reached 725.44 U/mL, and the TFC reached 2.38 mg/mL. FSBJ demonstrated strong antimicrobial activity against Escherichia coli, Staphylococcus aureus and Botrytis cinerea. Then, to investigate the antioxidant capacity of FSBJ, we used H2O2 to induce oxidative stress in C2C12 cells and assessed the protection conferred by FSBJ to damaged cells. It was discovered that after 24 h of treatment with FSBJ, not only was there an increase in the activities of intracellular SOD and glutathione peroxidase (GSH-Px), but also a reduction in reactive oxygen species (ROS) content, catalase (CAT) activity, and malondialdehyde (MDA) content. This research lays the theoretical groundwork and provides reference materials for the improved fermentation of sea buckthorn and demonstrates its resulting antioxidant effect.

Comparative quantitative analysis of fruit oil from Hippophae rhamnoides (seabuckthorn) by qNMR, FTIR and GC-MS / 中草药·英文版

OBJECTIVE@#To develop a qNMR method for quantitative analysis of triacylglycerols in fruit oil of Hippophae rhamnoides (seabuckthorn, SBT) and analyze commercial samples of SBT oils using GC-MS and FTIR.@*METHODS@#SBT fruit oil (IPHRFH) was extracted with hexane and the triglyceride (TAG) was isolated by vacuum liquid chromatography. Six different branded SBT oils purchased from e-commerce suppliers (Amazon) and in-house prepared SBT oil was analyzed by qNMR and fatty acyl composition of TAGs determined by using NMR. In-house oil was also analysed by GC-MS and FTIR spectroscopy.@*RESULTS@#The qNMR results showed that the oil contained 80.3% of triacylglycerol (TAG). The SBT oil TAGs comprised of linolenate 6.6%, palmitoleate/oleate 65.4%, and total saturated fatty acyl chain including palmitate 28% as determined by qNMR. GC-MS analysis revealed that the major acyl functionalities present in the TAG were palmitoleic acid 36.5%, oleic acid 12.9%, palmitic acid 21.2%, and linoleic acid 18%. Of the six commercial samples analyzed, samples from only one supplier (SW) were fruit oil; All others were the seed oils or mix of fruit oil and seed oil. The labels for samples except for the SW did not indicate whether it was fruit oil or seed oil.@*CONCLUSION@#The results suggest that SBT oil should be analyzed by combination of GC-MS, FTIR and qNMR for factual content of free fatty acid or TAGs, which are chemically different in nature and affect the quality of oil. GC-MS showed the content of omega free fatty acids after hydrolysis, while qNMR and FTIR showed the content of TAGs. The major acyl functionalities found in SBT fruit oil TAGs are palmitoleate/palmitate/oleate, while linoleate and linonelate make up a minor fraction. Furthermore, analysis of commercial samples showed discrepancies between label claims and actual content.

Assessment of Berries of Some Sea Buckthorn Genotypes by Physicochemical Properties and Fatty Acid Content of the Seed.

Sea buckthorn (Hippophae rhamnoides L.) is consumed mainly in its processed form. Therefore, the investigation of the physicochemical properties of its berries is a current task in the aspect of food processing. The aim of this study was to determine the physicochemical parameters (soluble solid content, total titratable acidity, sugar/acid ratio), color characteristics (L*, a*, b*) and fatty acid profile of five varieties ('Askola', 'Clara', 'Habego', 'Leikora', 'Mara') and one Hungarian candidate, R-01, to establish a basis for experiments on the processability of the whole berries (e.g., drying). The weight of the berry of 'Leikora' (0.64 g) was significantly higher than the other investigated fruits. The differences between the values of soluble solid content (6.3-10.84 °Brix) and titratable acid (1.4-3.7%) content of berries were significant. 'Mara' had the highest sugar/acid ratio. Regarding the fatty acid profile, the amount of unsaturated fatty acids was measured between 72.6-83.4%, including polyunsaturated fatty acids, which were between 32.3-58.1%. The seeds of the tested samples contained high concentrations of linoleic acid (17.0-33.2%) and linolenic acid (15.3-24.9%), mainly in the case of the 'Mara', 'Clara' and 'Askola' varieties. Candidate R-01 could be used as a raw material for functional foods due to its significant content of palmitoleic acid and a favourable omega-6/omega-3 ratio.

Evaluation of Antioxidative and Neuroprotective Activities of Total Flavonoids From Sea Buckthorn (Hippophae rhamnoides L.).

The aim of this study was to investigate the antioxidative and neuroprotective activities of total flavonoids from sea buckthorn (Hippophae rhamnoides L.) (TFH). Results indicated that TFH possessed DPPH radicals, hydroxyl radicals and superoxide anions scavenging activities. The neuroprotective potential was assessed with acetylcholinesterase (AChE) and monoamine oxidase A (MAO-A). The inhibition rates of AChE and MAO-A by 50 µg/ml TFH were 75.85 and 51.22%, respectively. The in vivo antioxidative and neuroprotective potential of TFH were explored in Caenorhabditis elegans. In the longevity assay, TFH (50 µg/ml) significantly increased the lifespan of wild-type C. elegans (29.40%). In the hydrogen peroxide-induced oxidative stress challenge, the antioxidant capacity of TFH-treated wild-type C. elegans was significantly enhanced. The C. elegans mutant strain CL4176 was used to study the neuroprotective effect of TFH in vivo. Results showed that TFH significantly delayed paralysis in C. elegans CL4176. Our study suggested total flavonoids from sea buckthorn (Hippophae rhamnoides L.) had the potential as an antioxidative and neuroprotective agent to extend aging and treat neurodegenerative diseases.

Protective effect of flavonoids extract of Hippophae rhamnoides L. on alcoholic fatty liver disease through regulating intestinal flora and inhibiting TAK1/p38MAPK/p65NF-κB pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: The therapeutic properties of Hippophae rhamnoides L. were already known in ancient Greece as well as in Tibetan and Mongolian medicine. Modern studies have indicated that Hippophae rhamnoides L. fermentation liquid protected against alcoholic fatty liver disease (AFLD). However, the underlying mechanism of Hippophae rhamnoides L. flavonoids extract (HLF) treating AFLD remains elusive. AIM OF THE STUDY: This study aimed to investigate the hepatoprotective effect of HLF in mice with AFLD and the interaction between AFLD and gut microbiota. MATERIALS AND METHODS: Chemical constituents of HLF were analyzed by Liquid Chromatography-Ion Trap-ESI-Mass Spectrometry. The Hepatoprotective effect of HLF was evaluated in mice with AFLD induced by alcohol (six groups, n = 10) daily at doses of 0.1, 0.2, and 0.4 g/kg for 30 consecutive days. At the end of experiment, mice were sacrificed and the liver, serum and feces were harvested for analysis. The liver histological changes were observed by H&E staining and oil red O staining. Moreover, the alterations of fecal microflora were detected by 16S rRNA gene sequencing. The inflammatory related genes were determined by qRT-PCR and western blotting respectively. RESULTS: The results showed that the oral administration of HLF remarkably alleviated hepatic lipid accumulation by decreasing the levels of ALT, AST, TG and TC. The levels of TNF-α, TGF-ß, and IL-6 were also reduced after treatment with HLF. Meanwhile, the protein and mRNA expression of NF-kB p65, MAPK p38 and TAK-1 in the liver of mice with AFLD were all reduced by HLF compared with model group. Furthermore, the 16S rRNA gene sequencing analysis demonstrated that HLF treatment can help restore the imbalance of intestinal microbial ecosystem and reverse the changes in Fimicutes/Bacterodietes, Clostridiales, Lachnospiraceae, S24-7, and Prevotella in mice with AFLD. CONCLUSION: HLF can effectively ameliorate liver injury in mice with AFLD, and regulate the composition of gut microbiota. Its regulatory mechanism may be related to TAK1/p38MAPK/p65NF-κB pathway. This study may provide novel insights into the mechanism of HLF on AFLD and a basis for promising clinical usage.

Total flavonoids of sea buckthorn (Hippophae rhamnoides L.) improve MC903-induced atopic dermatitis-like lesions.

ETHNOPHARMACOLOGICAL RELEVANCE: Sea buckthorn (Hippophae rhamnoides L.) is popularly used as a herbal medicine and food additive in the world. Total flavonoids of Hippophae rhamnoides (TFH) are reported to have anti-inflammatory and immunomodulatory activities. AIM: The effects of TFH on atopic dermatitis (AD)-like lesions induced by MC903 in mice was elucidated in the study. METHODS: To induce AD-like lesions, MC903 was adopted to apply repeatedly on the left ear in C57BL/6 mice. After induction of AD-like lesions, 0.5% and 1% TFH cream was applied topically on ears of mice once a day for 8 days. The degree of skin lesions was evaluated by macroscopical and histological methods. Expressions of filaggrin (FLG) was evaluated by Western blotting. Real-time polymerase chain reaction (qPCR) was adopted to detect the mRNA expression of thymic stromal lymphopoietin (TSLP), interferon (IFN)-γ, interleukin (IL-4), tumor necrosis factor (TNF)-α in skin lesions. In vitro, Cytokine Antibody Arrays were performed to measure production of cytokines in IFN-γ/TNF-α-treated HaCaT cells, Western blotting was employed to detect the expressions of p-NF-κB, p-ERK and p-P38. RESULTS: Topical application of TFH significantly improved the severity of dermatitis by inhibiting the infiltration of mast cell, increasing expression of FLG, decreasing the expressions of TNF-α, IL-4, IFN-γ and TSLP in skin lesions. TFH decreased the levels of IL-1α, IL-1ß, IL-6, monocyte chemoattractant protein (MCP)-1, MCP-3, macrophage-derived chemokine (MDC), platelet-derived growth factor (PDGF)-BB, thymus and activation regulated chemokine (TARC) in the supernatants of the HaCaT cells treated by IFN-γ/TNF-α. Furthermore, expressions of p-NF-κB, p-ERK and p-P38 were also decreased by TFH administration with dose dependent manner in HaCaT cells treated by IFN-γ/TNF-α. CONCLUSIONS: Topical application of TFH improved AD-like lesions in mice induced by MC903. Which exerted the effects of anti-inflammation and repairing skin barrier by regulating Th1/Th2 balance. This finding indicates that TFH is a novel potential agent for the external treatment of AD.

Oligo-FISH Can Identify Chromosomes and Distinguish Hippophaë rhamnoides L. Taxa.

Oligo-fluorescence in situ hybridization (FISH) facilitates precise chromosome identification and comparative cytogenetic analysis. Detection of autosomal chromosomes of Hippophaë rhamnoides has not been achieved using oligonucleotide sequences. Here, the chromosomes of five H. rhamnoides taxa in the mitotic metaphase and mitotic metaphase to anaphase were detected using the oligo-FISH probes (AG3T3)3, 5S rDNA, and (TTG)6. In total, 24 small chromosomes were clearly observed in the mitotic metaphase (0.89-3.03 µm), whereas 24-48 small chromosomes were observed in the mitotic metaphase to anaphase (0.94-3.10 µm). The signal number and intensity of (AG3T3)3, 5S rDNA, and (TTG)6 in the mitotic metaphase to anaphase chromosomes were nearly consistent with those in the mitotic metaphase chromosomes when the two split chromosomes were integrated as one unit. Of note, 14 chromosomes (there is a high chance that sex chromosomes are included) were exclusively identified by (AG3T3)3, 5S rDNA, and (TTG)6. The other 10 also showed a terminal signal with (AG3T3)3. Moreover, these oligo-probes were able to distinguish one wild H. rhamnoides taxon from four H. rhamnoides taxa. These chromosome identification and taxa differentiation data will help in elucidating visual and elaborate physical mapping and guide breeders' utilization of wild resources of H. rhamnoides.

Inhibitory effect of sea buckthorn extracts on advanced glycation endproduct formation.

This study shows the inhibitory effect of sea buckthorn (Hippophae rhamnoides L.) extracts, sea buckthorn leaf (HRL) and berry (HRB), on the formation of advanced glycation endproducts (AGEs), closely linked to diverse disease. In vitro assay revealed the superior inhibitory effect of HRL on the AGEs formation and AGEs-induced collagen crosslinking compared with that of HRB. Ultra-performance liquid chromatography-mass spectrometry results revealed that HRL displays a higher inhibition efficiency on the AGEs formation at 30 AGEs binding sites in bovine serum albumin than HRB. The high concentration of 3-sophoroside-7-rhamnoside in HRL compared with that in HRB may result in the strong inhibitory effect of HRL compared with that of HRB. HRL also exhibited significantly higher ABTS and DPPH radical scavenging activities than HRB. Overall, this study demonstrated that HRL has excellent potential as a dietary agent for controlling various diseases mediated by AGEs and oxidative stress.

Mechanochemical-Assisted Extraction and Hepatoprotective Activity Research of Flavonoids from Sea Buckthorn (Hippophaë rhamnoides L.) Pomaces.

Pomaces of sea buckthorn berry were usually side-products during the processing of juice. Due to a lack of an economical and effective extraction method, it was typically recognized as waste. For the purpose of resource utilization, the mechanochemical-assisted extraction (MCAE) method was applied to develop an ecofriendly extraction method and product with better pharmacology activity. The parameters were investigated through response surface methodology (RSM) design experiments. The processing conditions were optimized as follows: amount of Na2CO3 40%, ball-to-material rate 29:1 g/g, milling speed 410 rpm, milling time 24 min, extraction temperature 25 °C, extraction time 20 min and the solid-to-solution ratio 1:10 g/mL. Under these conditions, the yields of flavonoids from sea buckthorn pomaces were 26.82 ± 0.53 mg/g, which corresponds to an increase of 2 times in comparison with that extracted by the heat reflux extraction method. Meanwhile, the hepatoprotective activity of sea buckthorn pomaces extracts was studied by the liver injury induced by ip injection of tetracycline. Biochemical and histopathological studies showed that biomarkers in serum and liver of nonalcoholic fatty liver disease (NAFLD) mice were significantly ameliorated when sea buckthorn flavonoids extracted by MCAE were used. Altogether, these results demonstrate that, as a green and efficient extraction, MCAE treatment could increase the extraction yield of sea buckthorn flavonoids, meanwhile it could exhibit significant activity of improving liver function. This research provided a new way to use pomaces of sea buckthorn as a functional food. It also has great value on the comprehensive utilization of nature's resources.

Encapsulation of sea buckthorn (Hippophae rhamnoides L.) leaf extract via an electrohydrodynamic method.

Polyphenols from the leaves of sea buckthorn (Hippophae rhamnoides L.) are nutritious and bioactive substances that can be used as nutritional supplements. To improve their stability and bioaccessibility in vivo, chemical extracts of sea buckthorn leaves were, for the first time, encapsulated using electrohydrodynamic technology. The microcapsules were characterized using scanning electron microscopy, Fourier-transform infrared spectroscopy, and thermogravimetric analysis. The extract and microcapsules were evaluated for total phenols and flavonoids, total antioxidant activity, and their inhibitory effects on metabolic syndrome-related enzymes (α-glucosidase, α-amylase, and pancreatic lipase) under in vitro simulated digestion. The results indicated that the extract was successfully encapsulated; encapsulation protected polyphenols and flavonoids from degradation and increase their bioaccessibility in the intestine. The antioxidant activity and the inhibition of metabolic syndrome-related enzymes were better reserved after encapsulation. Our findings will help in promoting the potential of sea buckthorn as a nutritional supplement and expanding its commercial use.