Geographical equations of Swertia mussotii bioactivities: evidence from the western Sichuan region of China.

Swertia mussotii is the most authentic raw material used in Tibetan medicine in China for its various bioactivities. This natural medicine resource is at risk of being exhausted due to the double interference of climate change and anthropogenic over-collection. Little is known about habitat characteristics and the crucial environmental factors that influence the levels of active ingredients. The goal of this study is to understand the variability in the bioactive compound content of a wide range of wild S. mussotii as it adapts to changing environmental conditions. The target compound content of the whole plant material was analyzed with the environmental explanatory variables of the field sample sites using a constrained ordination method for their correlation analysis. The results show that 16.3 percent of the sampled wild S. mussotii populations with the highest bioactive content can be grouped into the elite type. The most prominent environmental variables affecting the content of major bioactive products include altitude, aspect, soil TK content, Fe content, and C/N and N/P ratios. Altitude and aspect put indirect effects that are mediated by plant height and density, N/P ratio puts a direct effect, while soil TK content, Fe content and C/N ratio have both direct and indirect effects on the bioactivity of S. mussotii. In addition to the total negative effects of altitude and C/N ratio, the remaining factors play a driving role. These findings demonstrate variation by geographical conditions across S. mussotii accessions for physiologic responses and secondary compounds in wild populations. The knowledge gained from this study can be used for environmental and plant physiology research, efficient collection of naturally active compounds, and conservation strategies for rare natural plant resources.

A rapid extraction process monitoring of Swertia mussotii Franch. With near infrared spectroscopy.

Swertia mussotii Franch. (SMF), a traditional Tibetan medicine, which has miraculous effect on treating hepatitis diseases. However, there is no research on its entire production process, and invisible production process has seriously hindered the SMF modern development. In this study, principal component analysis (PCA), subtractive spectroscopy, and two-dimensional correlation spectroscopy (2D-COS) were used to explain changes of characteristic groups in the extraction process. Four main characteristic peaks at 1884 nm, 1944 nm, 2246 nm and 2308 nm were identified to describe the changes of molecular structure information of total active components in SMF extraction process. In addition, multi critical quality attributes (CQAs) models were established by near-infrared spectroscopy (NIRS) combined with the total quantum statistical moment (TQSM). The coefficients of determination (R2eval and R2ival) were both greater than 0.99. The ratios of the standard deviation of validation to the standard error of the prediction (RPDe and RPDi) were greater than five. The quantitative model of AUCT could save time on primary data measurement by not requiring determination of indicator components compared with others. In conclusion, these results demonstrated that it was feasible to understand the SMF extraction process through AUCT and characteristic groups. These could realize the visual digital characterization and quality stability of the SMF extraction process.

Chemical profile of Swertia mussotii Franch and its potential targets against liver fibrosis revealed by cross-platform metabolomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Swertia mussotii Franch (SMF) is a well-known Tibetan medicine for the treatment of liver disease in China. However, the chemical profile and molecular mechanism of SMF against hepatic fibrosis are not yet well explored. AIM OF THE STUDY: This work aimed to elucidate the chemical profile of SMF and investigate the action mechanisms of SMF against carbon tetrachloride (CCl4)-induced hepatic fibrosis. MATERIALS AND METHODS: Ultra performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOFMS) and UNIFI platform was firstly employed to reveal the chemical profile of SMF. Cross-platform serum metabolomics based on gas chromatography/liquid chromatography-mass spectrometry were performed to characterize the metabolic fluctuations associated with CCl4-induced hepatic fibrosis in mice and elucidate the underlying mechanisms of SMF. Western blotting was further applied to validate the key metabolic pathways. RESULTS: A total of 31 compounds were identified or tentatively characterized from SMF. Twenty-seven differential metabolites were identified related with CCl4-induced liver fibrosis, and SMF could significantly reverse the abnormalities of seventeen metabolites. The SMF-reversed metabolites were involved in arachidonic acid metabolism, glycine, serine and threonine metabolism, tyrosine metabolism, arginine and proline metabolism, primary bile acid biosynthesis, glycerophospholipid metabolism and TCA cycle. The results of western blotting analysis showed that SMF could alleviate liver fibrosis by increasing the levels of CYP7A1, CYP27A1 and CYP8B1 and decreasing the level of LPCAT1 to regulate the metabolic disorders of primary bile acid biosynthesis and glycerophospholipid. CONCLUSION: It could be concluded that primary bile acid biosynthesis and glycerophospholipid metabolism were the two important target pathways for SMF-against liver fibrosis, which provided the theoretical foundation for its clinical use.

Exploring the Mechanism of Swertia mussotii Franch. for Hepatoprotective Effects with iTRAQ LC-MS/MS.

BACKGROUND AND OBJECTIVE: A Tibetan traditional herb named Swertia mussotii Franch., also called "Zangyinchen" by the local people of the Qinghai-Tibet area, has been used to protect the liver from injury for many years. However, the curative effect and molecular mechanism of the herb have not been demonstrated clearly. MATERIALS AND METHODS: In our study, serum alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels were examined after S. mussotii Franch. treatment of the acute liver injury on the carbon tetrachloride-induced rat model. Then, proteome analysis was conducted to explore the potential mechanism of SMT for hepatoprotective effects after iTRAQLC-MS/MS analysis (isobaric tag for relative and absolute quantification-liquid chromatography-mass spectrometer with tandem mass spectrometry). RESULTS: Serum results showed that alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels of rats with acute liver injury were all improved with SMT treatment. Moreover, proteome analysis suggested that, with S. mussotii Franch. treatment, the levels of lipid catabolic process and lipid homeostasis were all enhanced. Furthermore, the results of protein-protein interaction (PPI) analysis illustrated that the proteins assembled in PPI networks were found to be significantly enriched in response to lipid, negative regulation of lipase activity, and in response to lipopolysaccharides. Furthermore, the down-regulated MRP14 and MRP8 proteins were found to be involved in the lipid metabolism, which may indicate the mechanism of SMT for the protection of the liver from ALI induced by carbon tetrachloride. CONCLUSION: SMT herb could play a role in hepatoprotection and alleviating the acute liver injury by impacting the lipid metabolism associated with the biological process.

Exploration of the Hepatoprotective Effect and Mechanism of Swertia mussotii Franch in an Acute Liver Injury Rat Model.

BACKGROUND AND OBJECTIVE: Swertia mussotii Franch, also known as "Zangyinchen", is one of a Tibetan traditional herb used for treatment of liver diseases over thousands of years at Qinghai-Tibet Plateau, has been confirmed to be hepatoprotective. However, the underlying mechanism is largely unknown. MATERIALS AND METHODS: In this study, we evaluated the effect of S. mussotii treatment in a carbon tetrachloride-induced acute liver injury rat model by examining the serum alanine aminotransferase, aspartate aminotransferase, total bilirubin levels and performing histological observations of the liver tissues. Meanwhile, the metabolomics analysis was used to explore the molecular mechanism of S. mussotii treatment by high performance liquid chromatography tandem mass spectrometry. RESULTS: The results showed that S. mussotii treatment could effectively improve the serum alanine aminotransferase, aspartate aminotransferase, total bilirubin in acute liver injury rat model. Histological observation showed that S. mussotii treatment could effectively alleviate liver injury. Moreover, the metabolomics analysis showed that S. mussotii treatment could normalize the levels of many fatty acid metabolism related metabolites. And the results of pathway analysis showed that these metabolites significantly enriched in fatty acid biosynthesis pathway (myristic acid, dodecanoic acid and capric acid) and linoleic acid metabolism pathway (13-OxoODE). CONCLUSION: The results indicated that S. mussotii treatment could significantly improve acute liver injury through affecting the pathways related to lipid metabolism.

Cloning and bioinformatics analysis of SmMK gene in Swertia mussotii / 中草药

Objective: In order to identify the function of the mevalonate kinase (MK) which is a key enzyme of the mevalonate pathway (MVA) in Swertia mussotii, and to improve the study of MVA in S. mussotii. Methods: According to the SmMK gene sequence of transcriptome of S. mussotii, the specific primers were designed, the cDNA complete sequences was obtained by RT-PCR and the sequence was analyzed using bioinformatics. Prokaryotic expression vector MBP-SmMK was constructed and transformed into Escherichia coli Rosetta (DE3) for expression. Results: The results showed that SmMK cDNA complete sequences had a length of 1 164 bp encoding 387 amino acid residues. The SmMK protein shared high identity with other MK proteins of plants. And the protein signal peptide, transmembrane region, location, secondary, and tertiary structures were analyzed and forecasted. The SDS-PAGE results showed that the expressed proteins were consistent with the anticipated size, which was 40 970. Conclusion: This work will provide a foundation for research the SmMK protein functional and study MCA in S. mussotii. At the same time, it will supply the basis to improve the production of the isoprenoids.

Chemical constituents from Swertia mussotii Franch. (Gentianaceae).

The chemical investigation of ethanolic extract from Swertia mussotii Franch. has resulted in the isolation of 11 compounds which were identified as Orcinol-ß-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.

Cloning and expression of geranyl pyrophosphate synthase gene in Swertia mussotii / 中草药

Objective: To clone the geranyl pyrophosphate synthase gene from Swertia mussotii (SmGPPS), analyze the bioinformation of SmGPPS, and perform the gene expression. Methods: According to the SmGPPS gene sequence of transcriptome of S. mussotii, the specific primers were designed, the cDNA complete sequences was obtained by RT-PCR and the sequence was analyzed using bioinformatics. Prokaryotic expression vector pET-28a-SmGPPS was constructed and transformed into Escherichia coli BL-21 (DE3) for expression under 37℃ and induced by 1 mmol/L IPTG. The relative expression of gene SmGPPS in the leaf, stem, and flower of S. mussotii was also studied. Results: The results showed that SmGPPS cDNA complete sequences had a length of 1 119 bp encoding 372 amino acid residues. And the protein secondary and tertiary structures were analyzed and forecasted. The SmGPPS protein shared high identity with other GPPS proteins of plants. The SDS-PAGE results showed that the expressed proteins were consistent with the anticipated size. Relative RT-PCR analysis indicated that SmGPPS showed the highest transcript abundance in the leaf. Conclusion: This work will provide a foundation for further functional research of SmGPPS protein and increasing the product of iridoid compound by genetic engineering in S. mussotii.

Simultaneous determination of oleanolic acid, ursolic acid, quercetin and apigenin in Swertia mussotii Franch by capillary zone electrophoresis with running buffer modifier.

The method of capillary zone electrophoresis (CZE) with direct UV detection was developed for the determination of oleanolic acid, ursolic acid, quercetin and apigenin. and then for the first time successfully applied to the analysis of four analytes in Swertia mussotii Franch and its preparations. Various factors affecting the CZE procedure were investigated and optimized, and the optimal conditions were: 50 × 10(-3) mol/L borate-phosphate buffer (pH 9.5) with 5.0 × 10(-3) mol/L ß-cyclodextrin, 15 kV separation voltage, 20 °C column temperature, 250 nm detection wavelength and 5 s electrokinetic injection time (voltage 20 psi). Under the conditions, oleanolic acid, ursolic acid, quercetin and apigenin could be determined within the test ranges with a good correlation coefficient (r(2) > 0.9991). The limits of detection for conditions, oleanolic acid, ursolic acid, quercetin and apigenin were 0.3415, 0.2003, 0.0062 and 0.2538 µg/mL, respectively, and the intra- and inter-day relative standard deviations were no more than 4.72%. This procedure provided a convenient, sensitive and accurate method for simultaneous determination of oleanolic acid, ursolic acid, quercetin and apigenin in S. mussotii Franch.

Resources Survey on Rare and Endangered Medicinal Plant Swertia mussotii Franch / 世界科学技术-中医药现代化

The resource of rare and endangered medicinal plant Swertia mussotii Franch. in Tibet, Qinghai and Sichuan province were surveyed by ways of documents, interview, quadrat and market investigation. The results indi-cated that Swertia mussotii Franch. mainly distributed in Zuogong and Mangkang of Tibet, Yushu of Qinghai province, Shiqu, Daofu, Kangding, Maerkang, Jinchuan and Xiaojin of Sichuan province. According to the height above sea level, the distribution altitude was from 2 300 m (Kangding of Sichuan province) to 3 900 m (Mangkang of Tibet). There are distributions of Swertia mussotii Franch. within the scope of 2 600 m. The illumination, water, soil, temperature and altitude had significant influence on the distribution, growth and reserve of Swertia mussotii Franch. from different angles. In recent years, there was huge increase of market requirement in Swertia mussotii Franch. which were used in Tibetan medicine Zangyinc he n. Excess collection was the primary cause of rapid decreasing in resource of Swertia mussotii Franch.. It was suggested to strengthen the management of rational collection, as well as to accelerate the development of cultivation and production.

Study on Swertia mussotii Franch. by HPLC fingerprinting method combined with chemical pattern recognition approaches / 中国药学杂志

OBJECTIVE: To study the quality control method of Swertia mussotii Franch. METHODS: A high performance liquid chromatographic method was developed to establish the fingerprint of Swertia mussotii Franch. Nineteen samples from various batches were analyzed, furthermore, PCA were used to differentiate and evaluate the whole fingerprints. Some characteristic peaks were i-dentified preliminarily with HPLC-MS method. RESULTS: Eleven peaks were identified by MS. Swertiamarin and swertianolin were the key components for quality control. CONCLUSION: The method provides an academic reference for controlling the quality of Swertia mussotii Franch.

Chemical constituents from Swertia mussotii / 中草药

Objective: To study the chemical constituents of Swertia mussotii. Methods: Various chromatographic methods were employed to isolate the compounds and their structures were established by spectroscopic analysis. Results: Nineteen compounds were isolated from 75% ethanol extract of S. mussotii and identified as 1-hydroxyl-3, 4, 7, 8-tetramethoxyxanthone (1), 1, 7- dihydroxyl-3-methoxyxanthone (2), 1, 3, 7-trihydroxyxanthone (3), 1, 3, 7, 8-tetrahydroxyxanthone (4), 1, 3, 8-trihydroxyl-7- methoxyxanthone (5), 1, 3-dihyfroxyl-7, 8-dimethoxyxanthone (6), 1, 5, 8-trihydroxyl-3, 4-dimethoxyxanthone (7), 1-hydroxyl-3, 4, 5, 8-tetramethoxyxanthone (8), 1-hydroxyl-3, 5, 8-trimethoxyxanthone (9), (S)-(+)-gentiolactone (10a), (R)-(-)-gentiolactone (10b), 1, 8-dihydroxyl-3, 7-dimethoxyxanthone (11), 1-hydroxyl-3, 7, 8-trimethoxyxanthone (12), 1, 7-dihydroxyl-3, 8-dimethoxyxanthone (13), 1, 7, 8-trihydroxyl-3-methoxyxanthone (14), 1, 3, 5, 8-tetrahydroxyxanthone (15), 1, 7-dihydroxyl-3, 4, 8-trimethoxyxanthone (16), mangiferin (17), and oleanolic acid (18), respectively. Conclusion: Compounds 1-9 are firstly isolated from S. mussotii. Compounds 10a and 10b are firstly isolated from the plants in Swertia L.

Therapeutic effect of Swertia Mussotii Franch on immunological liver injury in mice / 西安交通大学学报(医学版)

Objective To observe the therapeutic effect of Swertia mussotii Franch is extracted by alcohol on immunological liver injury in mice,and further evaluate its immun-regulating effect in decreasing aminotransferase and protecting heptocytes. Methods BCG vaccine and lipopolysaccharide were intravenously injected to establish liver injury model.The effects of Swertia mussotii Franch on liver injury were contrasted. Results The high-,medium-and low-dose Swertia mussotii Franch groups which had been administered for 10 consecutive days,had significantly inhibited serum ALT and AST acticities of immunological liver injury;Swertia mussotii Franch could effectively prevent immunological liver injury by BCG+LPS. Conclusion Swertia mussotii Franch can protect mice from immunological liver injury and protect the function of liver.

Purification technology of iridoid glycosides and xanthones from Swertia mussotii by macroporous resin / 中草药

Objective To study the separation and purification technology of iridoid glycosides and xanthones from Swertia mussotii by macroporous resin.Methods The dynamic adsorption and desorption characteristics of macroporous adsorption resins HPD-300,HPD-400,HPD-600,AB-8,DM-301,and D-101-Ⅰ for swertiamarin and swertianolin were investigated.On the base of the investigation,the better macroporous adsorption resin was chosen to finish the experiments.Results The adsorption and elution characteristics of HPD-300 used to separate and purify iridoid glycosides and xanthones were very good.In the course of adsorption,the amount of used adsorption is 0.9 g dried medicinal herb/mL resin,with about 8 pH value of adsorption solution,and the volumetric rate is 2 BV/h; In the course of elution,the resin column chromatography was eluted gradiently with 7 BV 20% and 5 BV 70% EtOH after removing impurities with water.The elution rates were both more than 90%,and the contents of iridoid glycosides and xanthones were both up to 50% in the two dried fractions of 20% and 70% ethanolic elutions.Conclusion HPD-300 is an ideal resin with the best enrichment for separating and purifying both iridoid glucosioles and xanthanoes simultanously.