Ten trace elements and macro elements in Jin He Nao Xue Kang capsules, such as Cd, Co, Cr, Cu, Fe, Mg, Mn, Ni, Pb, and Zn were determined by flame atomic absorption spectrophotometry. The results showed that there are comparatively rich macro element Mg, and profitable elements such as trace elements Cu, Zn, Fe, Mn, Ni etc in Jin He Nao Xue Kang capsules. The contents of poisonous elements (Cd and Pb) are comparatively low. The content sequence of metal elements is as follow: Fe>Mg>Zn>Mn>Cu>Ni>Cr>Pb>Co>Cd. It provided useful data for discussing the relationship between trace elements and macro elements in Tibetan traditional medicine, and the cure for vascular and cerebral vascular disease.
In this paper, the varieties and origin of Primulaceae plants that used in Tibetan medicine were analyzed. The results showed that there were 3 genera and 44 species (including the varieties) of Primulaceae plants were recorded in the relevant literatures. Among them, 17 varieties were recorded in Tibetan names, 24 varieties were recorded in Chinese names and 1 variety was used in both of them. In current quality criteria of standards at all levels in China country, 6 varieties were recorded in Tibetan names and 6 original plants were involved, which were 35% and 14% of them respectively. Seventeen varieties were recorded in Chinese name and 7 original plants were involved, which were 30% and 16% of them respectively. In Tibetan medicine standards and literatures, there were big differences between Tibetan names and Chinese names which were translated from Tibetan names and its original plants. There were only regulations of morphological identification and microscopic authentication, so the standards were very inadequate. Therefore, through literatures research, resources and current situation investigations, combining the research and specification of the name and original of Tibetan medicine, the level of normalization and standardization could be enhanced, the stable and controllable safety and utility in clinical medication could be ensured to promote advancement of industry technology Tibetan medicine.
Background: Dracocephalum heterophyllum was a traditional Tibetan medicine possesses various pharmacological effects involved in anti-inflammatory, antibacterial activities. However, its anti-hepatitis, antioxidant activity and bioactive compounds have not been reported, the objective of this research work was to investigate the pharmacological activity and bioactive compounds of D. heterophyllum extracts. Results: In the present study, the anti-hepatics and antioxidant activities of four D. heterophyllum extracts (i.e. petroleum ether extracts, ethyl acetate extracts, n-BuOH extracts, and water extracts) were conducted. The main chemical constituent of petroleum ether and ethyl acetate extracts were also isolated using chromatographic techniques and identified by NMR spectroscopic methods. The anti-hepatitis assay showed that the petroleum ether and ethyl acetate extracts of D. heterophyllum significantly prolonged the mean survival times and reduced the mortality of mouse hepatitis model induced by concanavalin A (ConA). The levels of alanine transaminase, aspartate transaminase in blood serum could be decreased obviously by ethyl acetate extracts compared with ConA group (P < 0.01). The histological analysis demonstrated that the ethyl acetate extracts could inhibit apoptosis and necrosis caused by ConA. In addition, the antioxidant activities of the four extracts of D. heterophyllum were measured by DPPH assay, ABTS assay, anti-lipidperoxidation assay, ferric reducing antioxidant power assay, ferrous metal ions chelating assay and determination of total phenolic contents. The results showed that the ethyl acetate extract had the highest antioxidant activities, followed by petroleum ether extract. Finally, nine mainly compounds were isolated from the Petroleum ether and ethyl acetate extracts, including four triterpenes: oleanolic acid ( 1), ursolic acid ( 2), pomolic acid ( 3), 2α- hydroxyl ursolic acid ( 4), three flavonoids: apigenin-7- O-rutinoside ( 5), luteolin ( 8), diosmetin ( 9) and two phenolic acids: rosmarinic acid ( 6), methyl rosmarinate ( 7). Conclusion: The Ethyl acetate extract of D. heterophyllum had the highest anti-hepatitis and antioxidants activities, followed by petroleum ether extract. The bioactive substances may be triterpenes, flavonoids and phenolic acids, the ethyl acetate extracts of D. heterophyllum may be possible candidates in developing anti-hepatitis medicine.
Six compounds were isolated from an ethanol extract of Swertia mussotii and identified as 2-phenylethyl-β-D-glucoside (1), amaroswerin (2), 1,3,7,8-tetrahydroxyxanthone (3), swertiamarine (4), 1,3,8-trihydroxy-5-methoxyxanthone (5) and methylswertianin (6). Compounds 1, 2 and 6 were isolated from S. mussotii for the first time. The anti-inflammatory activities of the compounds were evaluated by determining their effect on the production of NO by LPS-stimulated RAW264.7 cells. Amaroswerin was the most potent inhibitor of NO release, with an IC50 value of 5.42 μg/mL. Treatment with amaroswerin inhibited expression of iNOS at both protein and mRNA levels. Amaroswerin also dose-dependently suppressed production of TNF-α, IL-6 and IL-1β and reduced expression of mRNA for these LPS-stimulated pro-inflammatory mediators. Amaroswerin thus inhibits the expression of iNOS, TNF-α, IL-6 and IL-1β by downregulating transcription in LPS-induced RAW264.7 macrophage cells, indicating that amaroswerin may be a valuable therapeutic agent for the treatment of inflammatory diseases.
Background: Hypecoum leptocarpum Hook. f. et Thoms., which is used in traditional Tibetan medicine as an antipyretic, antitussive, analgesic, and anti-inflammatory agent, contains a variety of alkaloids that could be responsible for its analgesic and anti-inflammatory properties. Objective: The present study was designed to investigate the anti-inflammatory activity of the total alkaloids from H. leptocarpum (AHL) in vitro and to elucidate the chemical structure of the anti-inflammatory components in AHL. Materials and Methods: Chemical characterization was performed using liquid chromatography/quadrupole-time-of-flight mass and diode-array detector-high performance liquid chromatography. The anti-inflammatory effects of AHL were investigated by measuring the production of inflammatory cytokines using enzyme-linked immunosorbent assay and mRNA expression by real-time polymerase chain reaction in lipopolysaccharide-induced RAW 264.7 macrophages. Results: Chemical analysis of AHL revealed the presence of seven alkaloids, protopine (13.3%), cryptopine (1.5%), leptopidinine, leptocarpine, corydamine, dihydroleptopine, and oxohydrastinine. AHL significantly suppressed the production of nitric oxide (NO), interleukin-1 beta (IL-1 β), IL-6, and tumor necrosis factor-alpha (TNF-α) in LPS-induced RAW 264.7 cells. The maximum levels of suppression of NO, IL-1 β, IL-6, and TNF-α were 86.8% ± 2.2%, 70.1% ± 1.5%, 100.1% ± 2.5%, and 50.8% ± 3.6%, respectively. IC50values of suppression of cytokine production by AHL were 7.47 ± 2.81 μg/mL (NO), 0.12 ± 0.28 μg/mL (IL-1 β), 0.56 ± 0.37 μg/mL (IL-6), and 18.95 ± 5.23 μg/mL (TNF-α). AHL was also shown to downregulate mRNA expression of inducible NO synthase, IL-1 β, IL-6, and TNF-α in vitro. Conclusion: The study provides convincing evidence that AHL has strong anti-inflammatory activity. The potent activity is likely a result of synergy between the different alkaloids. Abbreviations used: The total alkaloids from H. leptocarpum: AHL; Nitric oxide: NO; Interleukin-1 beta IL-1β; Interleukin-6: IL-6; Tumor necrosis factor-alpha: TNF-α; Prostaglandin E2: PGE2; Inducible nitric oxide synthase: iNOS; Nonsteroidal anti-inflammatory drugs: NSAIDs; lipopolysaccharide: LPS; The total ion chromatograms: TIC; The liquid chromatography/quadrupole-time of flight: LC/Q-TOF; Nuclear factor-kappa B: NF-κB; Janus kinase-signal transducers and activators of transcription: JAK-STAT. [ABSTRACT FROM AUTHOR]
As important secondary plant metabolites, naphthoquinones exhibit a wide range of biological activities. However, their potential as sustainable alternatives to synthetic acaricides has not been studied. This study for the first time investigates the acaricidal activity of naphthoquinones against Psoroptes cuniculi in vitro. Furthermore, the in vivo activity, the skin irritation effects, the cytotoxicity and the inhibitory activities against mite acetylcholinesterase (AChE) and glutathione S-transferase (GST) of the two compounds that displayed the best insecticidal activity in vitro were evaluated. Among fourteen naphthoquinones and their analogs, juglone and plumbagin were observed to possess the strongest acaricidal activities against P. cuniculi with LC50 values of 20.53 ppm and 17.96 ppm, respectively, at 24 h. After three treatments, these two chemicals completely cured naturally infested rabbits in vivo within 15 days, and no skin irritation was found in any of the treated rabbits. Compared to plumbagin, juglone presented no or weak cytotoxicity against HL-7702 cells. Moreover, these two chemicals significantly inhibited AChE and GST activity. These results indicate that juglone has promising toxicity against P. cuniculi, is safe for both humans and animals at certain doses, and could be used as a potential alternative bio-acaricide for controlling the development of psoroptic mange in agricultural applications.
The chemical investigation of ethanolic extract from Swertia mussotii Franch. has resulted in the isolation of 11 compounds which were identified as Orcinol-β-D-glucoside (1), Shamimin (2), Mangiferin (3), Decussatin (4), Bellidifolin (5), Desmethylbellidifolin (6), Protocatechuic acid (7), 1,7-Dihydroxy-3,8-dimethoxyxanthone (8), 1,8-Dihydroxy-3,5-dimethoxyxanthone (9), 1-Hydroxy-3,5-dimethoxyxanthone (10), Telephioidin (11). The chemical structures of these compounds were identified by a combination of spectroscopic analysis and a comparison with those reported in literature. Among them, compounds 1, 2, 7 and 11 were isolated from the genus Swertia for the first time. Moreover, the chemotaxonomic significance of these compounds was summarised. The chemotaxonomic study suggests that there is a close chemotaxonomic relationship between S. mussotii and other species of Swertia, such as S. punicea, S. macrosperma, S. japonica, S. phragmitiphylla, S. chirayita, S. cordata and S. binchuanensis, with presence of compounds 3~6, 8~10. The xanthones and their glycosides may sever as important chemotaxonomic markers of Swertia genus.
A phytochemical investigation on the aerial parts of a Tibetan medicine Meconopsis horridula, by solvent extraction, repeated chromatographies on silica gel, Sephadex LH-20, and preparative TLC techniques, led to the isolation of 9 compounds. By spectroscopic analysis and comparison of its 1H and 13C-NMR data with those in literatures, their structures were identified as oleracein E(1), N-( trans-p-coumaroyl) tyramine (2), chrysoeriol (3), apigenin (4), hydnocarpin (5), p-coumaric acid glucosyl ester (6), stigmast-5-ene-3beta-ylformate (7), 3beta-hydroxy-7alpha-ethoxy-24beta-ethylcholest-5-ene (8), and beta-sitosterol (9), respectively, among which compounds 6-8 were isolated from the genus for the first time,and 1,3 were isolated from the species for the first time. A MTT method was applied to evaluate the cytotoxic activity of compounds 14 against the human hepatocellular liver carcinoma cell line (HepG2), and compound 1 showed significant cytotoxicity against HepG2,with its inhibitory rate of 52.2% at 10 micromol x L(-1).;
The study aims at providing a new suitable way to promote artificial cultivation, solving the problem of resources increasingly endangered wild medicine, and protecting the wild resources of Tibetan medicine. The content of quercetin,kaempferol and isorhamnetin was determined by HPLC. The correlation between flavonoids components and ecological factors was analyzed using partial least-squares regression (PLSR). Based on Maxent model combining using ArcGIS software, suitable regionalization for H.rhamnoides subsp. sinensis was studied.The results showed that the difference of quercetin,kaempferol and isorhamnetin content in samples from different regions were obvious. The main factors effecting quercetin content accumulation were the altitude andthe average monthly precipitation in January and August. The main factors effecting kaempferol accumulation were the altitude andthe average monthly precipitation in the coldest quarter and December. The main factors effecting isorhamnetin accumulation were the average monthly precipitation in August, January and the coldest quarter.The regional distribution suitability index for H.rhamnoides subsp. sinensis was 0-0.708. The suitable area 590 500 km², accounting for 6.13% of the total area. The preferably suitable area was 552 500 km², accounting for 5.73% of the total area.The methods used in the study is simple and feasible, the result is reliable which provide a new approach for Tibetan medicine resources sustainable exploitation and utilization.
OBJECTIVE: To establish a method for determination of 10 ingredients such as gentiopicroside, sweroside, and mangiferin in India swertia, and settle the index components and their limits.METHOD: By Welch materials AQ-C18 column, determination was conducted by the gradient elution with methanol and 0.4% formic acid as mobile phase, with column temperature 30 degrees C, flow rate at 1.0 mL x min(-1), and 254 nm as the detection wavelength.
RESULT: The linear relatives of 10 ingredients were good. The method showed the high precision and good reproducibility, and recovery rates were between 97% and 103%. The ingredients of market com-modities varied greatly.
CONCLUSION: This method is simple, sensitive, reproducible, and applicable to the determination of the main ingredients in India Swertia. Sweroside and mango glycosides were suggested as the index components for determination in Jia Di (Swertia chirayita), and their content limits are not less than 0.1%, 0.3%, respectively.
OBJECTIVE: To establish the HPLC fingerprint for Halenia elliptica herbs, a traditional Tibetan medicine, in order to study constituents contained in H. elliptica from different habitats and compare their differences.METHOD: HPLC analysis was made on a Welchrom-C18 (4.6 mm x 250 mm, 5 microm) with water and acetonitrile as mobile phase. The wavelength was detected as 265 nm, the flow rate was 1.0 mL x min(-1), and the column temperature was 40 degrees C. The software for chromatographic fingerprint was applied to analyze the similarity. And principal component analysis was conducted.
RESULT: Twelve common chromatographic peaks were identified by fingerprint, showing a low similarity in constituent and variety. The significant difference in the proportion between xanthones and aglycones in each batch of herbs indicated no notable correlation between constituent characteristics and geographic locations of habitats.
CONCLUSION: The method is so simple, exclusive, stable and highly repeatable that it can provide reference for identification and quality assessment of H. elliptica herbs.
* Mindful learning is applied into the domain of environmental protection. * We test the association between mindful learning and pro-environmental behavior in the perspective of self-expansion. * Mindful learning could influence self-reported pro-environmental behavioral intentions in both positive and negative ways. * The mindful-leaning material need to be carefully selected and employed.; This current article explores the differential effects of mindful learning on pro-environmental behavior from the perspective of self-expansion. A total of 253 participants were recruited for four experiments. In Study 1, the mindful-learning group reported greater levels of pro-environmental behavioral intentions compared to a randomized control. In Study 2, we utilized different learning materials focusing on self, humans, or the biosphere in three sub-experiments. Study 2a manipulated mindsets by a self-related focus and revealed participants in a mindfulness condition had lower pro-environmental behavioral intentions than those in the mindlessness group. Study 2b centered on humans and results showed that participants in a mindfulness group reported higher levels of pro-environmental behavioral intentions. Finally, Study 2c induced mindsets with a biospheric focus, showing participants in the mindful-learning condition had greater pro-environmental behavioral intentions. Combined, the studies provide empirical evidence that mindful learning could influence self-reported pro-environmental behavioral intentions both positively and negatively.
* Mindful learning is applied into the domain of environmental protection. * We test the association between mindful learning and pro-environmental behavior in the perspective of self-expansion. * Mindful learning could influence self-reported pro-environmental behavioral intentions in both positive and negative ways. * The mindful-leaning material need to be carefully selected and employed.; This current article explores the differential effects of mindful learning on pro-environmental behavior from the perspective of self-expansion. A total of 253 participants were recruited for four experiments. In Study 1, the mindful-learning group reported greater levels of pro-environmental behavioral intentions compared to a randomized control. In Study 2, we utilized different learning materials focusing on self, humans, or the biosphere in three sub-experiments. Study 2a manipulated mindsets by a self-related focus and revealed participants in a mindfulness condition had lower pro-environmental behavioral intentions than those in the mindlessness group. Study 2b centered on humans and results showed that participants in a mindfulness group reported higher levels of pro-environmental behavioral intentions. Finally, Study 2c induced mindsets with a biospheric focus, showing participants in the mindful-learning condition had greater pro-environmental behavioral intentions. Combined, the studies provide empirical evidence that mindful learning could influence self-reported pro-environmental behavioral intentions both positively and negatively.
ObjectiveTo investigate the effects of short-term forest bathing on human health.
Methods
Twenty healthy male university students participated as subjects and were randomly divided into two groups of 10. One group was sent on a two-night trip to a broad-leaved evergreen forest, and the other was sent to a city area. Serum cytokine levels reflecting inflammatory and stress response, indicators reflecting oxidative stress, the distribution of leukocyte subsets, and plasma endothelin-1 (ET-1) concentrations were measured before and after the experiment to evaluate the positive health effects of forest environments. A profile of mood states (POMS) evaluation was used to assess changes in mood states.
Results
No significant differences in the baseline values of the indicators were observed between the two groups before the experiment. Subjects exposed to the forest environment showed reduced oxidative stress and pro-inflammatory level, as evidenced by decreased malondialdehyde, interleukin-6, and tumor necrosis factor a levels compared with the urban group. Serum cortisol levels were also lower than in the urban group. Notably, the concentration of plasma ET-1 was much lower in subjects exposed to the forest environment. The POMS evaluation showed that after exposure to the forest environment, subjects had lower scores in the negative subscales, and the score for vigor was increased.
Conclusion
Forest bathing is beneficial to human health, perhaps through preventive effects related to several pathological factors.
To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.
OBJECTIVE: To evaluate the medicinal reasonableness and resource utilization of Dida from different species.METHOD: With common characteristic absorption peaks of HPLC fingerprints and SPSS cluster, the composition similarity of Dida from different species was evaluated.
RESULT: The composition similarity of HPLC fingerprints of 33 Dida samples from 15 species and 1 variety originated from Swertia, Halenia, Gentianopsis, Lomatogonium was difference. The original species can be clustered into four groups by the relative area of 10 common characteristic peaks of HPLC fingerprints. The compositions of four different genera are quite different.
CONCLUSION: Because of containing iridoids, xanthones, and triterpenes which have liver protection and cholagogue functions, all of species from Swertia, Halenia, Gentianopsis and Lomatogonium in Gentianaceae are classified as Dida in Tibetan medicine. According to the composition difference among different species, the HPLC fingerprints established for Dida from different source are an effective means to identify nd control the quality of Dida.
Context: Tsothel, a traditional Tibetan medicine, is regarded as 'the king of essences'. Nevertheless, tsothel has aroused serious concern regarding its biosafety because its main component is HgS. Unfortunately, toxicological studies on tsothel are scarce. Objective: As inorganic mercury has high affinity for the kidney, the present investigation was designed to determine the potential nephrotoxicity and mechanism of tsothel. Materials and methods: Sprague-Dawley rats were orally administered different doses of tsothel (0, 66.70, 33.35 and 16.68 mg/kg) daily for 180 days, followed by the withdrawal of tsothel for 120 days. Then, the related nephrotoxicity was examined by the ICP-MS, ELISA, colorimetric, RT-PCR, HE staining, immunohistochemical staining and flow cytometry methods. Results: Although tsothel administration led to a large accumulation of Hg (794.25 ± 464.30 ng/g in the 66.70 mg/kg group, 775.75 ± 307.89 ng/g in the 33.35 mg/kg group and 532.60 ± 356.77 ng/g in the 16.68 mg/kg group) in the kidney after 120 days of tsothel withdrawal, the blood CREA and BUN, urinary Kim-1, NAG, RBP and β2-MG, renal SOD, MDA, pathology, proliferation, apoptosis and cell cycle had no significant changes compared with the control group. Additionally, the high GSH content (318.87 ± 44.19 nmol/mL in the 33.35 mg/kg group) and the relative expression levels of Kim-1 (1.08 ± 0.11 in the 33.35 mg/kg group), MT-1 (1.46 ± 0.10 in the 66.70 mg/kg group, 1.61 ± 0.19 in the 33.35 mg/kg group and 1.57 ± 0.14 in the 16.68 mg/kg group) and GST-Pi (1.76 ± 0.89 in the 33.35 mg/kg group) mRNA recovered to normal after tsothel withdrawal. Interestingly, the change trend of GST-Pi gene expression was consistent with the change trend of GSH activity. Conclusions: Overall, our study shows that tsothel administration did not induce overt nephrotoxicity but did have reversible stress-related effects. These results suggest that tsothel affects stress response mechanisms with the involvement of detoxifying enzyme systems. The formulation method and chemotype could play a role in the reduced toxicity potential of tsothel compared to common mercurials. [ABSTRACT FROM AUTHOR]
CONTEXT: Tsothel, a traditional Tibetan medicine, is regarded as 'the king of essences'. Nevertheless, tsothel has aroused serious concern regarding its biosafety because its main component is HgS. Unfortunately, toxicological studies on tsothel are scarce. OBJECTIVE: As inorganic mercury has high affinity for the kidney, the present investigation was designed to determine the potential nephrotoxicity and mechanism of tsothel. MATERIALS AND METHODS: Sprague-Dawley rats were orally administered different doses of tsothel (0, 66.70, 33.35 and 16.68 mg/kg) daily for 180 days, followed by the withdrawal of tsothel for 120 days. Then, the related nephrotoxicity was examined by the ICP-MS, ELISA, colorimetric, RT-PCR, HE staining, immunohistochemical staining and flow cytometry methods. RESULTS: Although tsothel administration led to a large accumulation of Hg (794.25 ± 464.30 ng/g in the 66.70 mg/kg group, 775.75 ± 307.89 ng/g in the 33.35 mg/kg group and 532.60 ± 356.77 ng/g in the 16.68 mg/kg group) in the kidney after 120 days of tsothel withdrawal, the blood CREA and BUN, urinary Kim-1, NAG, RBP and β2-MG, renal SOD, MDA, pathology, proliferation, apoptosis and cell cycle had no significant changes compared with the control group. Additionally, the high GSH content (318.87 ± 44.19 nmol/mL in the 33.35 mg/kg group) and the relative expression levels of Kim-1 (1.08 ± 0.11 in the 33.35 mg/kg group), MT-1 (1.46 ± 0.10 in the 66.70 mg/kg group, 1.61 ± 0.19 in the 33.35 mg/kg group and 1.57 ± 0.14 in the 16.68 mg/kg group) and GST-Pi (1.76 ± 0.89 in the 33.35 mg/kg group) mRNA recovered to normal after tsothel withdrawal. Interestingly, the change trend of GST-Pi gene expression was consistent with the change trend of GSH activity. CONCLUSIONS: Overall, our study shows that tsothel administration did not induce overt nephrotoxicity but did have reversible stress-related effects. These results suggest that tsothel affects stress response mechanisms with the involvement of detoxifying enzyme systems. The formulation method and chemotype could play a role in the reduced toxicity potential of tsothel compared to common mercurials.
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.
The ITS2 barcode was used toidentify Tibetan medicine "Dida", and tosecure its quality and safety in medication. A total of 13 species, 151 experimental samples for the study from the Tibetan Plateau, including Gentianaceae Swertia, Halenia, Gentianopsis, Comastoma, Lomatogonium ITS2 sequences were amplified, and purified PCR products were sequenced. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner V3.7.1. The Kimura 2-Parameter (K2P) distances were calculated using MEGA 6.0. The neighbor-joining (NJ) phylogenetic trees were constructed. There are 31 haplotypes among 231 bp after alignment of all ITS2 sequence haplotypes, and the average G±C content of 61.40%. The NJ tree strongly supported that every species clustered into their own clade and high identification success rate, except that Swertia bifolia and Swertia wolfangiana could not be distinguished from each other based on the sequence divergences. DNA barcoding could be used as a fast and accurate identification method to distinguish Tibetan medicine "Dida" to ensure its safe use.
Ethnopharmacological relevance: Saussurea laniceps Hand.-Mazz. (SL) has long been used under the herbal name Tibetan 'Snow Lotus' for the treatment of rheumatoid arthritis, stomachache and dysmenorrhea in Tibetan folk medicine. Since herbal medicine (HM) is a synergistical system with multiple components, both of the metabolism and pharmacokinetic studies of HM are interdependent. This study aimed to develop an integrated strategy based on the UPLC-DAD-QTOF-MS technique for metabolism and pharmacokinetic studies of HM.Material and methods: SL was used here as a test herb to verify the feasibility of the proposed strategy. SL was administered to rats, then, the blood plasma, urine and feces were analyzed to determine the metabolic profiles. Using our strategy, umbelliferone and scopoletin were evaluated to be the key bioactive components. Their pharmacokinetic parameters were measured and biotransformation pathways were elucidated.Results: After oral administration of SL to rats, 17 components in blood, 10 components in urine and 2 components in feces were identified and characterized using our UPLC-DAD-QTOF-MS method. Umbelliferone, scopoletin and their metabolites were found to be the major components involved in the metabolism process. Literature reports also suggest that umbelliferone and scopoletin are responsible for the therapeutic effects of SL, thus these two components were selected as the active markers for pharmacokinetic study. In the test of validity, the established method presented good linearity with R-2> 0.99. The relative standard deviation value was below 13.9% for precision, and recovery studies for accuracy were found to be within the range 91.8-112.5%.Conclusion: The present strategy offers, simultaneously, precision in quantitative analysis (metabolism study) and accuracy in quantitative analysis (pharmacokinetic study) with greater efficiency and less costs, which is therefore reliably used for integrated metabolism and pharmacokinetic studies of HM. (C) 2014 Elsevier Ireland Ltd. All rights reserved.
Antioxidant potencies of an ethanolic extract and its components from <i>Lepidium latifolium</i> were investigated. In this study, we found that the ethyl acetate soluble fraction of <i>L. latifolium</i> was a rich source of antioxidant, resulting from its high total phenolic content. To determine the antioxidant components of the ethyl acetate fraction, a bioassay-guided fractionation approach using 1,1-diphenyl-2-picrylhydrazyl, 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulfonate), and ferric reducing antioxidant power assays were conducted. Nine compounds were isolated and their structures were identified by MS and NMR spectral data and comparison to reported data. They are Quercetin-3-O-β-<i>d</i>-sophoroside-7-O-α-<i>l</i>-rhamnoside (<i>1</i>), Apetalumoside B6 (<i>2</i>), Kaempferol-3-O-β-<i>d</i>-glucopyranosyl-(1-2)-β-<i>d</i>-glucopyranoside-7-O-β-<i>d</i>-glucopyranoside (<i>3</i>), Kaempferol-7-O-α-<i>l</i>-rhamnopyranoside (<i>4</i>), Kaempferol-3-O-β-<i>d</i>-glucopyranoside-7-O-α-<i>l</i>-rhamnopyranoside (<i>5</i>), Kaempferol-3-O-(2-O-feruloyl-β-<i>d</i>-glucopyranosyl-(1-2)-β-<i>d</i>-glucopyranoside)-7-O-glucopyranoside (<i>6</i>), Kaempferol-3-O-β-<i>d</i>-sophoroside-7-O-α-<i>l</i>-rhamnoside (<i>7</i>), Kaempferol-3-O-robinoside-7-O-(2″″-(E)-feruloyl)-sophoroside (<i>8</i>), Quercetin-3-O-(2,6-di-O-β-<i>d</i>-glucopyranosyl)-β-<i>d</i>-glucopyranoside-7-O-α-<i>l</i>-rhamnopyranoside (<i>9</i>), compounds <i>1</i>, <i>2</i>, <i>4</i>, and <i>8</i> had potent free radical scavenging activity. The IC<sub>50</sub> values of these compounds were 9.8-12.3 and 7.4-31.4 μg/mL in DPPH and ABTS assays, respectively. The results indicate that <i>L. latifolium</i> is a potential natural source of antioxidants to treat several diseases related to oxidant by-products of human metabolism.
Swertia mussotii Franch. and Swertia chirayita Buch.-Ham. have been commonly used under the same name "Zangyinchen" for the treatment of liver and gallbladder diseases in traditional Tibetan medicine. Detailed characterization and comparison of the complete set of metabolites of these two species are critical for their objective identification and quality control. In this study, a rapid, simple and comprehensive (1)H NMR-based metabolomics method was first developed to differentiate the two species. A broad range of metabolites, including iridoid glycosides, xanthones, triterpenoids, flavonoids, carbohydrates, and amino acids, were identified. Statistical analysis showed evident differences between the two species, and the major markers responsible for the differences were screened. In addition, quantitative (1)H NMR method (qHNMR) was used for the target analysis of the discriminating metabolites. The results showed that S. mussotii had significantly higher contents of gentiopicrin, isoorientin, glucose, loganic acid, and choline, whereas S. chirayita exhibited higher levels of swertiamarin, oleanolic acid, valine, and fatty acids. These findings indicate that (1)H NMR-based metabolomics is a reliable and effective method for the metabolic profiling and discrimination of the two Swertia species, and can be used to verify the genuine origin of Zangyinchen.
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