The genus Tripterygium: A phytochemistry and pharmacological review.

The genus Tripterygium belongs to the family Celastraceae, and contains three species, i.e. Tripterygium wilfordii Hook. F, Tripterygium hypoglaucum (Levl.) Hutch. and Tripterygium regelii Sprague et Takeda. All three species are reported to have excellent medicinal properties that help to cure rheumatoid arthritis, nephrotic syndrome, systemic lupus erythematosus and widely used as a folk medicine in China. Phytochemical studies have led to discovering more than 500 secondary metabolites in this genus, including five main types: sesquiterpenoids, diterpenes, triterpenoids, flavonoids, lignans. This work provides structurally grouping statistic of 198 secondary metabolites of Tripterygium species published from 2008 to the present, as well as pharmacological knowledges in the past five years. The information will be helpful for developing the new discoveries of medicinal value related to the genus Tripterygium.

[Identification and quality evaluation of Tripterygium wilfordii].

With the extensive clinical application of Tripterygium wilfordii, there are many counterfeit products on the market. Traditional technology can not effectively identify the authenticity of the traditional Chinese medicine. Therefore, a strategy of accurate identification and quality evaluation of Tripterygium based on DNA barcode and chemical fingerprint spectrum was established. Based on DNA barcode technology, HMMer annotation method of hidden Markov model and K2P model were used to analyze genetic distance.BLAST1, nearest distance and phylogenetic tree (NJ-tree) methods were used to assess the identification efficiency of the ITS2 barcode. The fingerprint of 27 T. wilfordii was established by UPLC-PDA method, and the similarity of the fingerprint of different sources was evaluated. The main components of T. wilfordii were determined by LC-MS/MS. The results revealed that the intraspecific genetic distances of T. wilfordii were lower than the interspecific genetic distances between T. wilfordii and its adulterants. The results of similarity search showed that ITS2 sequence was used to identify T. wilfordii and its adulterants. The clustering of T. wilfordii and its adulterants was clear in the tree of NJ cluster, and 12 of 27 samples were identified as true T. wilfordii.The chemical fingerprint spectrum research indicates that the feature one region can distinguish the false product of tripterygium glycosides more intuitively. The cluster analysis of HCA-thermal map showed that the contents of six active components of T. wilfordii from different habitats were significantly different, which could be used to evaluate the quality of T. wilfordii. This paper is of guiding significance for the accurate identification and quality evaluation of Tripterygium medicinal plants.

Investigation on Species Authenticity for Herbal Products of Celastrus Orbiculatus and Tripterygum Wilfordii from Markets Using ITS2 Barcoding.

Herbal material is both a medicine and a commodity. Accurate identification of herbal materials is necessary to ensure the safety and effectiveness of medication. With this work, we initiated an identification method to investigate the species authenticity for herbal products of Celastrus orbiculatus and Tripterygum wilfordii utilizing DNA barcoding technology. An ITS2 (internal transcribed spacer two) barcode database including 59 sequences was successfully established to estimate the reliability of species-level identification for Celastrus and Tripterygium. Our findings showed that ITS2 can effectively and clearly distinguish C. orbiculatus, T. wilfordii and its congeners. Then, we investigated the proportions and varieties of adulterant species in the herbal markets. The data from ITS2 region indicated that 13 (62%) of the 21 samples labeled as "Nan-she-teng" and eight (31%) of the 26 samples labeled as "Lei-gong-teng" were authentic; the remaining were adulterants. Of the 47 herbal products, approximately 55% of the product identity were not in accordance with the label. In summary, we support the efficacy of the ITS2 barcode for the traceability of C. orbiculatus and T. wilfordii, and the present study provides one method and reference for the identification of the herbal materials and adulterants in the medicinal markets.

[Studies on difference of chemical compositions in plant species of Tripterygium genus].

We studied the content of chemical compositions and correlation among species of Tripterygium genus by principal component analysis(PCA) and variance analysis(ANOVA), and we also studied the difference among the 3 species.Using [BMIm]PF6 ionic liquid-based ultrasonic-assisted extraction, we determined the contents of 11 compounds including wilforgine, wilforzine, triptophenolide, wilforine, triptoquinone A, triptolide, tripterin, egallocatechin, epigallocatechin, catechin, and epicatechin in 28 batches of the Tripterygium species by HPLC and PCA. Partial least squares analysis (PLS) and ANOVA were also performed to verify the results.The analysis results of PCA and PLS showed that three species of Tripterygium genus were clustered into three regions respectively, and triptoquinone A was the important factor which affected the aggregation of these three species.There was a significant difference among the contents of 11 chemical components in the three species(P<0.000 1).These results indicated that there was a certain correlation between the chemical compositions and the classification of the species, and the difference of the chemical compositions among the three species was obvious. In this work, the content determination method is rapid and accurate, and the analysis method is simple and convenient, which provides a reference for the classification, the efficacy and the toxicity of the species.

Molecular Analysis of Chinese Celastrus and Tripterygium and Implications in Medicinal and Pharmacological Studies.

Celastrus and Tripterygium species, which are used in traditional Chinese medicine, have attracted much attention due to their anti-tumor promoting and neuroprotective activities, in addition to their applications in autoimmune disorders. However, systematic relationships between them and among species are unclear, and it may disturb their further medicinal utilization. In the present study, the molecular analysis of combined chloroplast and nuclear markers of all Chinese Celastrus and Tripterygium was performed, and clear inter- and intra-genus relationships were presented. The result suggests that Tripterygium constitute a natural monophyletic clade within Celastrus with strong support value. Fruit and seed type are better than inflorescence in subgeneric classification. Chinese Celastrus are classified for three sections: Sect. Sempervirentes (Maxim.) CY Cheng & TC Kao, Sect. Lunatus XY Mu & ZX Zhang, sect. nov., and Sect. Ellipticus XY Mu & ZX Zhang, sect. nov. The phylogenetic data was consistent with their chemical components reported previously. Owing to the close relationship, several evergreen Celastrus species are recommended for chemical and pharmacological studies. Our results also provide reference for molecular identification of Chinese Celastrus and Tripterygium.

Identification of Species in Tripterygium (Celastraceae) Based on DNA Barcoding.

Species of genus Tripterygium (Celastraceae) have attracted much attention owing to their excellent effect on treating autoimmune and inflammatory diseases. However, due to high market demand causing overexploitation, natural populations of genus Tripterygium have rapidly declined. Tripterygium medicinal materials are mainly collected from the wild, making the quality of medicinal materials unstable. Additionally, identification of herbal materials from Tripterygium species and their adulterants is difficult based on morphological characters. Therefore, an accurate, convenient, and stability method is urgently needed. In this wok, we developed a DNA barcoding technique to distinguish T. wilfordii HOOK. f., T. hypoglaucum (LÉVL.) HUTCH, and T. regelii SPRAGUE et TAKEDA and their adulterants based on four uniform and standard DNA regions (internal transcribed spacer 2 (ITS2), matK, rbcL, and psbA-trnH). DNA was extracted from 26 locations of fresh leaves. Phylogenetic tree was constructed with Neighbor-Joining (NJ) method, while barcoding gap was analyzed to assess identification efficiency. Compared with the other DNA barcodes applied individually or in combination, ITS2+psbA-trnH was demonstrated as the optimal barcode. T. hypoglaucum and T. wilfordii can be considered as conspecific, while T. regelii was recognized as a separate species. Furthermore, identification of commercial Tripterygium samples was conducted using BLAST against GenBank and Species Identification System for Traditional Chinese Medicine. Our results indicated that DNA barcoding is a convenient, effective, and stability method to identify and distinguish Tripterygium and its adulterants, and could be applied as the quality control for Tripterygium medicinal preparations and monitoring of the medicinal herb trade in markets.

[Different metabolites of leaves between Tripterygium wilfordii and Tripterygium hypoglaucum based on UPLC-Q-TOF-MS].

To analysis the differences between Tripterygium wilfordii and T. hypoglaucum, specimens of their leaves were collected from five production regions and analyzed by ultra performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS). The data were analyzed by multivariate statistical method, such as hierarchical cluster analysis (HCA) principal component analysis (PCA) and orthogonal signal correction partial least square discrimination (OPLS-DA). Potential markers with VIP values above 5.0 and corresponding r values above 0.85, were selected and further tested by combining mann-Whitney nonparametric. Those with P < 0.001 and AUC = 1 were confirmed as metabolite markers to discriminate them from each other. Results revealed that the two species were obviously different in their leaf metabolites. Based on their mass spectra, 23 potential metabolite markers were identified to distinguish T. wilfordii from T. hypoglaucum.

[Investigation Report of Tripterygium wilfordii and Tripterygium hypoglaucum].

OBJECTIVE: To get the information of resources, cultivation, commodity circulation and other aspects of Tripterygium wilfordii and Tripterygium hypoglaucum. METHODS: Collect samples in 13 locations of Tripterygium wilfordii and Tripterygium hypoglaucum, compare their plant morphological characteristics and growth habit, and investigate their wild resources conditions, planting information, easy-confused varieties and different commodity features. RESULTS: (1) Tripterygium wilfordii and Tripterygium hypoglaucum were mainly collected under woods or on the edge of woods,and light and moisture attributed to their distribution to some extent. (2) Wild resources of Tripterygium wilfordii and Tripterygium hypoglaucum were shrinking, and both of their cultivation history were relatively short and their cultivation technique were still in a low level. (3) Due to lack of harvesting and processing standards, decoction pieces, varying from roots, rhizomes to stems of plants, were all sold as commercial medicines. CONCLUSION: Wild resources of Tripterygium wilfordii and Tripterygium hypoglaucum are shrinking,and the standardized research on cultivation-harvest processing and commercial medicines remains to be further carried out.

[Cloning and expression analysis of a acetyl-CoA U-acetyltransferase gene (TwAACT) from Tripterygium wilfordii].

In this study, based on the transcriptome data, we cloned the full-length cDNAs of TwAACT gene from Tripterygium wilfordii suspension cells, and then analyzed the bioinformation of the sequence, detected the genetic differential expression after being induced by methyl jasmonate (MeJA) by RT-PCR. The full-length cDNA of the TwAACT was 1 704 bp containing a 1 218 bp open reading frame (ORF) encoding a polypeptide of 405 amino acids (GeneBank accession No. KP297934). The deduced isoelectric point (pI) was 6.10, a calculated molecular weight was about 41.20 kDa, and online prediction showed that TwAACT had two catalytic active sites. After the induction of MeJA, the relative expression level of TwAACT increased rapidly. The expression level of TwAACT was highest at 24 h. TwAACT was cloned firstly, that laid the foundation for identifying thegene and illustrating thebiosynthesis mechanism and its synthetic biology.

[RAPD analysis for genetic relationship and diversity of three species of genus Tripterygium].

OBJECTIVE: To study the genetic relationship and diversity of 3 species which belong to Tripterygium. METHOD: Samples were collected and divided into 4 types: typical T. wilfordii, typical T. hypoglaucum and their middle type according to morphological characters, and T. regelii. RAPD markers were used to measure the genetic relationship and diversity of 110 individuals from 22 natural populations in China. RESULT AND CONCLUSION: 10 primers were selected from 100 ones screened. A total of 128 bands were scored and 123 of them were polymorphic. Cluster analysis indicated that all the samples could be divided into 3 parts: 5 individuals from T. regelii gathered closely and were separated from other population and formed single branch. All 4 populations from T. hypoglaucum gathered directly. The middle type showed a nearer relationship with T. wilfordii than with T. hypoglaucum, although there was genetic differentiation between populations from middle type and those from T. wilfordii. The middle type has more diversity than T. wilfordii. The distance between different middle type populations and T. wilfordii was different. The existence of middle type populations between T. wilfordii and T. hypoglaucum suggested that the 2 species should be combined into one species. Considering all populations except T. regeli, the generic differentiation among populations was significant and the genetic diversity existed mainly among different populations. So samples of Tripterygii should be collected from different ppopulations for the reservation of the genetic diversity.

Medicinal chemistry and pharmacology of genus Tripterygium (Celastraceae).

Plants in the genus Tripterygium, such as Tripterygium wilfordii Hook.f., have a long history of use in traditional Chinese medicine. In recent years there has been considerable interest in the use of Tripterygium extracts and of the main bioactive constituent, the diterpene triepoxide triptolide (1), to treat a variety of autoimmune and inflammation-related conditions. The main mode of action of the Tripterygium extracts and triptolide (1) is the inhibition of expression of proinflammatory genes such as those for interleukin-2 (IL-2), inducible nitric oxide synthase (iNOS), tumor necrosis factor-alpha (TNF-alpha), cyclooxygenase-2 (COX-2) and interferon-gamma (IFN-gamma). The efficacy and safety of certain types of Tripterygium extracts were confirmed in human clinical trials in the US and abroad. Over 300 compounds have been identified in the genus Tripterygium, and many of these have been evaluated for biological activity. The overall activity of the extract is based on the interaction between its components. Therefore, the safety and efficacy of the extract cannot be fully mimicked by any individual constituent. This review discusses the biochemical composition and biological and pharmacological activities of Tripterygium extracts, and their main bioactive components.