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.

Research progress on chemical constituents and pharmacological activities of seabuckthorn and prediction of its Q-markers / 中国中药杂志

Seabuckthorn contains flavonoids, tannins, terpenoids, polysaccharides, and vitamins, which have anti-inflammation,anti-oxidation, liver protection, anti-cardiovascular disease, anti-aging, immune enhancing, anti-tumor, and anti-bacterial activities.We reviewed the papers focusing on the chemical constituents, pharmacological activities, and utilization of seabuckthorn. The quality markers(Q-markers) of seabuckthorn were predicted and analyzed based on original plant phylogeny, chemical composition correlation, traditional medicinal properties, pharmacodynamic correlation, traditional and extended efficacy, pharmacokinetics, metabolic processes, and measurable components. With this review, we aim to provide theoretical reference for the quality control and quality standard establishment of seabuckthorn, so as to promote the rational exploitation and utilization of seabuckthorn resources, and improve the healthy and sustainable development of seabuckthorn industry.

Regulation of seabuckthorn seed oil on metabolic pathway disturbances on rats exposed to chronic unpredictable mild stress / 中草药

Objective: To analyze and explore the changes of the metabolites in serum and urine of the chronic unpredictable depression rat model (CUMS) rats disturbed by seabuckthorn seed oil with using nuclear magnetic resonance spectroscopy (1H-NMR) and KEGG database. Methods: CUMS was prepared and the rats seabuckthorn seed oil was ig given to intervene at the same time for four weeks. Then the serum and the urine of rats were collected to test by 1H-NMR based metabolomics approach which purpose was to analyze the changes of the metabolites. Finally, the KEGG database was searched to analyze metabolic pathway. Results: A total of 17 metabolites were identified as potential biomarkers in serum and urine for the CUMS model, of which nine metabolites were found in serum including higher levels of lipids, β-OH-butyrate, lactate, and N-acetyl glycoprotein and lower levels of alanine, betaine, trimethylamine N-oxide, α-glucose, and β-glucose and eight metabolites were found in urine including level increments for succinate, aspirate and level reductions for acetate, 2-oxoglutaric acid, citrate, TMA, betaine, and phenylalanine, compared with the negative control group (NS). After intervened by seabuckthorn seed oil, the above different metabolites have appeared different degree of callback, closing to NS. Conclusion: These metabolic responses to CUMS treatment by seabuckthorn seed oil seemed to be some alterations in energy metabolism, lipid metabolism, amino acid metabolism, methylamine metabolism, and synthesis of neurotransmitter.

Rapid determination of six flavonoids from seabuckthorn leaves by RP-HPLC-DAD / 中国药科大学学报

To establish a rapid determination method of six flavonoids:catechin,rutin,myricetin,quercetin,kaempferol and isorhamnetin,from seabuckthorn leaves by RP-HPLC-DAD.The seabuckthorn leaves were first degreased by petroleum ether,extracted by ethanol,and determined by RP-HPLC-DAD.The six flavonoids were separated and eluted by a Shimadzu C18(150 mm ×2.1 mm,5 μm) column with methanol-water (0.1％ phos phoric acid) (60∶ 40) at a flow rate of 1.0 mL/min.The detection wavelength were as follow:catechin 208 nm,rutin 257 nm,myricetin 373 nm,quercetin 371 nm,kaempferol 367 nm,and isorhamnetin 371 nm,respectively.The injection volume was 20 μL.The contents of the six flavonoids were in the range of 0.47 to 30.00 μ,g/mL with good linearity.The validation of the method,including precision,stability and recovery rate,was acceptable.The established method can be used for fast determination of the content of six flavonoids in seabuckthorn leaves.

Digital gene expression profiling of dendritic cells treated with Seabuckthorn favones / 中华微生物学和免疫学杂志

Objective To study the changes in morphology , phenotypes and gene expression pro-files of dendritic cells (DCs) following treatment with Seabuckthorn flavones (SF).Methods DCs were treated with 200μg/ml of SF and then cultured for 7 days.Changes in the morphology of DCs were observed under light microscope .Flow cytometry was used to detect DC surface molecules .Total RNA was extracted to construct the library for digital gene expression profiling ( DGE ) .Differentially expressed genes were screened out and further analyzed by gene ontology ( GO) enrichment analysis and Kyoto encyclopedia of genes and genomes ( KEGG ) pathway enrichment analysis .Results Compared with control group , SF treatment significantly enhanced the expression of HLA-DR, CD80, CD83 and CD86 on DCs.A total of 355 differentially expressed genes were screened out by DGE , including 176 up-regulated genes and 179 down-regulated genes .GO enrichment was mainly involved in the regulation and development of the immune sys -tem and other biological processes .KEGG pathway analysis showed that the significantly enriched pathways were closely related to inflammation , the immune system, cancer and other diseases .Conclusion SF can promote the expression of DC co-stimulatory molecules and pro-mature molecules, and regulate the expres-sion of immunity-related genes such as CD11a, SLAMF6, LMCD1, TSC22D3 and IKZF3.

Inhibitory effects of seabuckthorn polysaccharide on oxidative stress in mice with actue liver injury and modulatory effect on BCL-2/Bax and PPAR-γexpression / 中国免疫学杂志

Objective:To explore the inhibitory effects of seabuckthorn polysaccharide on hepatic oxidative stress in a mice model of acute liver injury induced by intraperitoneal injection of LPS and D -GalN and detect the expression on hepatic BCL-2/Bax and PPAR-γ.Methods: C57BL/6 male mice were randomly divided into six groups:control group ( CTRL), model group ( L/G), dexamethasone positive control group ( DXM ) , low ( SPL ) , medium ( SPM ) and high dose group ( SPH ) of seabuckthorn polysaccharide.Mice in the SPL,SPM and SPH group were gavaged with 50,100 and 200 mg/kg seabuckthorn polysaccharide for 14 days respectively.Acute liver injury model were established by intraperitoneal injection of LPS (10 μg/kg) and D-GalN (700 mg/kg) .Serum and liver samples were collected 4 h after model establishment .Serum levels of ALT and AST and the content of MDA were de-tected.Hepatic expression of SOD 2 BCL-2 and Bax was determined by Western blot and the expression of PPAR-γwas detected by im-munohistochemistry .Results:ALT and AST levels significantly increased in the model group and decreased dose-dependently after pre-treatment with seabuckthorn polysaccharide .The level of MDA in the model group increased significantly as compared with the control group and decreased in seabuckthorn polysaccharide groups ,while the level of SOD 2 decreased in the model group and recovered in sea-buckthorn polysaccharide groups .The expression of Bax decreased after pretreatment with seabuckthorn polysaccharide .There was no obvious effect on BCL-2 expression after sea buckthorn polysaccharide supplementation .The expression of PPAR-γreduced in the sea-buckthorn polysaccharide group as compared with the model group .Conclusion:Seabuckthorn polysaccharide protects against LPS /D-GalN-induced liver injury.The effect is associated with an upregulation of SOD 2 and downregulation of Bax .

Rapid analysis of compounds in leaves of Chinese seabuckthorn and Tibetan seabuckthorn by UPLC/Q-TOF-MS / 中国中药杂志

The study is aimed to analyze the chemical components in leaves of Chinese seabuckthorn and Tibetan seabuckthorn qualitatively and compare the differences between them by using ultra performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS).The chromatographic separation of the components was achieved ona Waters ACQUITY UPLC-T3 column (2.1 mm×100 mm, 1.7 μm）using gradient mobile phase consisting of acetonitrile (A) and aqueous solution (B). The identification of the separated compounds was performed on atandem mass spectrometry (MS/MS)by fragmentation patterns under the negative electrospray ionization. The parameters of ion source were as follows：capillary voltage, 2 000 V; Cone voltage, 40 V. The ion source temperature, 100 ℃; collision gas argon; sheath gas flow rate, 900 L•h⁻¹; sheath gas temperature, 450 ℃. Through the analysis of mass spectrometry data and with the help of literature data, a total of 35 compounds were detected and most of them were flavonoids. Among these compounds, 29 were common components for the two species, two components were unique to Chinese seabuckthorn and 4 were characteristic components of Tibetan seabuckthorn. The results indicated that the compositions of the two kinds of seabuckthorn leaves were quite similar. It is also demonstrated that UPLC/Q-TOF-MS method could be applied to rapidly and effectively analyze and speculate the compounds in leaves of Chinese seabuckthorn and Tibetan seabuckthorn.

Genetic and chemical discrimination of traditional Tibetan medicine seabuckthorn based on DNA barcode and ¹H-NMR metabolic method / 中国中药杂志

To differentiate three medicinal Hippopahe species of seabuckthorn, a combined genetic and chemical identification method was established in this study. ITS2 and psbA-trnH were tested for identification of 3 species of seabuckthorn. Detection of the kimura 2-parameter (K2P) distance, the neighbor-joining (NJ) tree and the barcoding gap were used to assess the identification efficiency. ¹H-NMR based metabolic method was applied to acquire the profile of metabolites. PCA was used to analysis the metabolite data. The results indicated that DNA barcode combined ¹H-NMR based metabolic method is a powerful tool for the identification of 3 medicinal Hippopahe species of seabuckthorn. The finding demonstrated that different genetic variation and chemical constituents existed among 3 medicinal Hippopahe species of seabuckthorn. The combined identification method will improve the reliability of species discrimination and could be applicable to much other ethnic medicine which has various origins in China.

Study on HPLC fingerprint and quantitative analysis of 3 components of Tibetan medicine Gyantse Seabuckthorn / 中国中药杂志

This study is to establish an HPLC fingerprint and quantitative analysis of 3 components of Gyantse Seabuckthorn from different producing areas.The separation was developed on Shimadzu InertSustain C18column (4.6 mm × 250 mm,5 μm) by gradient elution with acetonitrile and 0.2% phosphoric acid water as mobile phase at a flow rate of 1.0 mL•min ⁻¹; the detection wavelength was set at 360 nm and column temperature was set at 30 ℃. The data calculation was performed with similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine(Version 2004A).The fingerprints of 10 batches of Gyantse Seabuckthorn were carried out by similarity comparison, and 12 chromatographic peaks were extracted as the common peaks of fingerprint, of which three main active ingredients were successfully determined. This is the first established fingerprint and multi-component quantitative determination of Gyantse Seabuckthorn by using HPLC. This method has good precision stability and repeatability that could provide basis for quality control and evaluation of Gyantse Seabuckthorn.

The effect of seabuckthorn flavonoids against ultraviolet injury / 重庆医学

Objective Through the UVB rays radiation skin injury model ,the study investigated the effect of seabuckthorn flavonoids on the protection and restoration of the skin ,in order to provide experimental basis of looking for the new drugs against ultraviolet injury .Methods By UVB radiating repeatedly ,the study built UVB radiation skin injury model in mice ,to observed the morphological changes of the skin in every groups of experimental animal ,the super oxide dismutase (SOD) activity of the skin and the peroxidation damage degree of the skin lipid ,which is also the content changes of malondialdehyde (MDA) and collagen .Results The change of model group whose epidermis layer increased and the dermis thickened was more obvious than the normal group ,so the UVB radiation skin injury model was built successfully .The model group compared with the normal group ,its SOD activity was significantly lower and its MDA content was significantly higher (P< 0 .01) .However ,the SOD activities of positive control group and being fed with seabuckthorn flavonoids group were significantly higher and their MDA contents were significantly lower (P<0 .01) .The model group compared with the normal group ,its collagen content was significantly lower (P< 0 .05) ,and the collagen contents of positive control group and being fed with seabuckthorn flavonoids group were higher ,whose difference was extremely significant(P< 0 .01) .However ,the positive control group had hyperplastic scar .Conclusion Seabuckthorn flavonoids had the fol‐lowing effects :preventing the UVB repeating radiation damage to the skin ,improving the SOD activity of skin tissue and the colla‐gen content ,inhibiting the increase of MDA content and scar hyperplasia ,and having antagonism effect of UVB causing mice skin photoaging .

Effect of seabuckthorn extract on scopolamine induced cognitive impairment.

Present study involves evaluation of effects of 75% ethanolic extract of seabuckthorn [Hippophae rhamnoides L. (SBT)] leaves on scopolamine induced cognitive impairment in rats using three different oral doses i.e. 50, 100 and 200 mg/kg body weight through assessment of various biochemical and behavioural parameters. Scopolamine administration resulted in an increase in acetylcholinesterase (AChE) activity (approximately 9% with respect to the control group) and malonaldehyde (MDA) content. The increased AChE activity was significantly reduced in animals receiving 200 and 100 mg/kg of SBT extract. Animals treated with SBT extract showed significantly reduced MDA level in all the doses. This reduction in MDA content indicates that SBT leaf extract has potent antioxidant activities and exhibits a protective effect against oxidative damage induced by scopolamine. Behavioural studies also indicated significant improvement. The results suggest that SBT leaf extract has potential effects against scopolamine induced cognitive impairment by regulating cholinergic marker enzyme activity (AChE activity) and promoting the antioxidant system and may be explored for its use in cognitive disorders.

Effect of seabuckthorn extract on delayed chlorophyll fluorescence on Cd and Co ions treated wheat seedlings.

The protecting effect of “Hypporamine PL” compound isolated from dry leaves of seabuckthorn (Hippophae rhamneides L.) on photosystem 2 (PS 2) activity suppression induced by CdCl2 and CoCl2 treatment in the 7 day old wheat seedlings (Triticum aestivum L.) under different pH of growth medium was investigated by measurement of millisecond delayed fluorescence (ms-DF) of chlorophyll intact leaves. The value o-i/p-s of ms-DF ratio was reduced under the Cd2+ and Co2+ treatments on 60 and 65% respectively at pH 6.7. Acidification of medium (pH 5.0) results in decreasing of ratio o-i/p-s only approximately on 30% in average. In the alkaline medium the lowering of o-i/p-s on 41% is observed in both ions treatments. This decreasing of o-i/p-s ratio occurred due to decreasing of fast phase o-i amplitude. At the same time the widening and increasing of slow phase p-s amplitude was observed. The compound “Hypporamine PL” limited the decrease of ms-DF components induced by heavy metals. It is suggested that the protective effect of “Hypporamine PL” on the photochemical reactions in the PS2 is due to catechins, epicatechins, quercetin and other polyphenols, containing in this compound, preventing the free radicals formation in the PS2 under treatment by heavy metal ions.

Study on the Quality Control of Compound Seabuckthorn Oil Granule / 中国药房

OBJECTIVE:To establish the quality control method for compound seabuckthorn oil granule.METHODS:Radix astragali,angelica root,rhizoma chuanxiong,seabuckthorn seed oil were identified by TLC and the content of astragalosideⅠin compound seabuckthorn oil granule was determined by TLC-scanning method.RESULTS:Radix astragali,angelica root,rhizoma chuanxiong,seabuckthorn seed oil could be detected by TLC.Good liner relationship achieved when the sample application size of astregalosideⅠwas with the range of 0.98?g～4.90?g,the average recovery was 99.62%(RSD=2.51%).CONCLUSION:This method is simple,accurate with good reproducibility and strong specificity,and it can be used for the quality control of compound seabuckthorn oil granule.

Study on the Actions of Seabuckthorn Oil Emulsion Against Chronic Atrophic Gastritis and Gastric Ulcer in Rats / 中国药房

OBJECTIVE:To study the actions of seabuckthorn oil emulsion against chronic atrophic gastritis and gastic ulcer in rats.METHODS:Chronic atrophic gastritis and gastric ulcer models were established in rats and mice and the antiulcer and anti-chronic atrophic gastritis actions of seabuckthorn oil emulsion were determined.RESULTS:Seabuckthorn oil emulsion could obviously prevent gastric ulcer and chronic atrophic gastritis induced by deoxycholic acid and stress and could decrease the index of acute gastric ulcer caused by ligature of pylorus in rats and could oppose the ulcer induced by indomethacin-ethanol significantly in mice.CONCLUSION:The results suggest that the antiulcer and anti-chronic atrophic gastritis actions of seabuckthorn oil emulsion may be related with its protection of the gastric mucosal barrier and neutralization of acid.

Comparative study of Sea buckthorn pulp oil with its seed oil on anti-gastric ulcer action and its mechanism / 中成药

Objective: To compare anti-gastric ulcer action of sea buckthorn pulp oil(SBPO) with sea buckthorn seed oil(SBSO) and to explore its mechanism. Methods : The anti-gastric ulcer actions of SBPO and SBSO were observed and compared on the gastric ulcer induced by water immersion stress, reserpine, pylorus ligation and acetic acid. The effects on gastric secretion in rats were studied with pylorus ligation. The effects on gastric emptying were observed by methyl orange remaining rate in stomach of mouse. The analgesic effect was tested by the hot-plate method and writhing method. Results : SBPO and SBSO markedly inhibited gastric ulcer induced by water immersion stress, reserpine and pylorus ligation and promoted the healing of the gastric ulcer induced by acetic acid. SBPO and SBSO significantly inhibited the secretion of gastric acid and pepsin and promoted the secretion of gastric mucous. SBPO and SBSO delayed gastric emptying in normal mouse. SBPO and SBSO significantly decreased the number of twisting body in mice. Conclusion : Both SBPO and SBSO could have anti-gastric ulcer effect with similar potency. The mechanism of anti-gastric ulcer might be their effect of inhibition of the secrection of gastric juice, promotion of the secrection of gastric mucous, inhibition of activity of gastric smooth muscle and analgetic effect.

Effects of Tiangui menopause capsules on rat ovarian granulosa cell apoptosis / 中国药理学通报

Aim To observe effects of Seabuckthorn fatty acids on old rat ovarian granulosa cell apoptosis and expression of bcl-2 and fas,then discuss the adjustive effect of Seabuckthorn fatty acids on ovarian function of menopause.Methods 22 months old rats were randomly divided into 5 groups,estrogen control group,old control group,Seabuckthorn fatty acids group of high dose,middle dose and low dose.In addition,one young control group was made up of 3 months old rats.Rat ovarian granulosa cell apoptosis was detected by TUNEL.The expression of protein bcl-2 and protein fas were detected by immunohistochemistry,and bcl-2 mRNA and fas mRNA by in situ hybridization(ISH).Grey analyse was done using Q550CW image analyse system.Results Compared with the old rats control group,Seabuckthorn fatty acids of high dose,middle dose and low dose can differently restrain rat ovarian granulosa cell apoptosis,enhance the expression of protein bcl-2 and bcl-2 mRNA,weakened the expression of protein fas and fas mRNA.Conclusion Seabuckthorn fatty acids restrains ovarian granulosa cell apoptosis through adjusting the expression of bcl-2 and fas.Sequentially,it adjusts estrogen level and treats menopause syndrome.