[Hypolipidemic effect of Coptis chinensis and C. deltoidea on type 2 diabetes rats through SCAP/SREBP-1c signal pathway].

The type 2 diabetes rat model was induced with high-sugar and high-fat diet combined with low-dose STZ. This study investigated the hypolipidemic mechanism of Coptis chinensis and C. deltoidea. After 30 days of administration, HOMA-IR and the content of TG in serum were detected, and the expressions of SCAP, SREBP-1c were tested by the method of Western blot and Real-time PCR analysis. The test results showed that both components can significantly alleviate insulin resistance and down-regulate the expressions of SREBP-1c and SCAP in liver tissue of type two diabetes mellitus. Compared with the control group, there were significant differences in relevant protein expression (P<0.05, P<0.01). This indicates that the inhibition of SREBP-1c and SCAP expressions may be the hypolipidemic mechanism of Coptidis Rhizoma on type 2 diabetes mellitus. The results also showed that C. deltoidea has a better efficacy in lipid elimination, but a weaker hypoglycemic effect against C. chinensis.

Biogeography of Coptis Salisb. (Ranunculales, Ranunculaceae, Coptidoideae), an Eastern Asian and North American genus.

BACKGROUND: Numerous studies have favored dispersal (colonization) over vicariance (past fragmentation) events to explain eastern Asian-North American distribution patterns. In plants, however the disjunction between eastern Asia and western North America has been rarely examined using the integration of phylogenetic, molecular dating, and biogeographical methods. Meanwhile, the biogeographic patterns within eastern Asia remain poorly understood. The goldthread genus Coptis Salisb. includes 15 species disjunctly distributed in North America, Japan, mainland China, and Taiwan. We present a dated phylogeny for Coptis under the optimal clock model and infer its historical biogeography by comparing different biogeographic models. RESULTS: The split of Coptis and Xanthorhiza Marshall occurred in the middle Miocene (ca. 15.47 Ma). Coptis started their diversification in the early late Miocene (ca. 9.55 Ma). A late Miocene vicariance event resulted in the eastern Asian and western North American disjunction in the genus. Within eastern Asia, dispersals from mainland Asia to Japan and from Japan to Taiwan occurred at ca. 4.85 Ma and at ca. 1.34 Ma, respectively. CONCLUSIONS: Our analyses provide evidence that both vicariance and dispersal events have played important roles in shaping the current distribution and endemism of Coptis, likely resulting from eustatic sea-level changes, mountain formation processes and an increasing drier and cooler climate from the middle Miocene onwards.

[Analysis on genetic diversity and genetic relationship of medicinal species in Coptis by SCoT].

The genetic diversity and genetic relationship among four medicinal species of Coptis were detected by the approach of sequence-related amplified polymorphism (SCoT). The associated genetic parameters were calculated by POPGENE1.31. The systematic diagram of genetic relationship were clustered by TREECONW. The results showed that a total of 434 bands were produced by using 28 primers, of which 430 were polymorphic loci. There was a high level of genetic diversity among species (PPB=99.1%,Na=1.990 6,Ne=1.329 3,H=0.212 2,I=0.337 8). However, genetic diversity was lower within species, the average of genetic parameters wasPPB=16.8%,Na=1.168 2, Ne=1.073 0,H=0.043 7,I=0.067 7. The Nei's genetic differentiation coefficient was 0.794 0, that indicated that most of the genetic variation existed among species. By clustering analysis, different individuals gathered in the same group. The results confirmed that SCoT marker can be used as one of the effective methods to reveal the genetic diversity and relationship among medicinal species of Coptis.

The First Comprehensive Phylogeny of Coptis (Ranunculaceae) and Its Implications for Character Evolution and Classification.

Coptis (Ranunculaceae) contains 15 species and is one of the pharmaceutically most important plant genera in eastern Asia. Understanding of the evolution of morphological characters and phylogenetic relationships within the genus is very limited. Here, we present the first comprehensive phylogenetic analysis of the genus based on two plastid and one nuclear markers. The phylogeny was reconstructed using Bayesian inference, as well as maximum parsimony and maximum likelihood methods. The Swofford-Olsen-Waddell-Hillis and Bayesian tests were used to assess the strength of the conflicts between traditional taxonomic units and those suggested by the phylogenetic inferences. Evolution of morphological characters was inferred using Bayesian method to identify synapomorphies for the infrageneric lineages. Our data recognize two strongly supported clades within Coptis. The first clade contains subgenus Coptis and section Japonocoptis of subgenus Metacoptis, supported by morphological characters, such as traits of the central leaflet base, petal color, and petal shape. The second clade consists of section Japonocoptis of subgenus Metacoptis. Coptis morii is not united with C. quinquefolia, in contrast with the view that C. morii is a synonym of C. quinquefolia. Two varieties of C. chinensis do not cluster together. Coptis groenlandica and C. lutescens are reduced to C. trifolia and C. japonica, respectively. Central leaflet base, sepal shape, and petal blade carry a strong phylogenetic signal in Coptis, while leaf type, sepal and petal color, and petal shape exhibit relatively higher levels of evolutionary flexibility.

Quality evaluation and species differentiation of Rhizoma coptidis by using proton nuclear magnetic resonance spectroscopy.

Rhizoma coptidis, a broadly used traditional Chinese medicine, derives from the dried rhizomes of Coptis chinensis Franch, Coptis deltoidea C.Y. Cheng et Hsiao and Coptis teeta Wall. Quantitative determination of protoberberine alkaloids in R. coptidis is critical for controlling its quality. In this study, a rapid, simple and accurate quantitative (1)H NMR (qNMR) method was developed for simultaneous determination of berberine, jatrorrhizine, epiberberine, coptisine, palmatine and columbamine in R. coptidis from the three species. Method validation was performed in terms of selectivity, precision, repeatability, stability, accuracy, robustness and linearity. The average recoveries obtained were in the range of 96.9-102.4% for all the six alkaloids. In addition, the qNMR data were analyzed with analysis of variance (ANOVA), hierarchical clustering analysis (HCA) and principal component analysis (PCA), and the results showed that the contents of the active alkaloids have significant difference among the three species. Compared with the conventional HPLC approach, the proposed qNMR method was demonstrated to be a powerful tool for quantifying the six alkaloids due to its unique advantages of high robustness, rapid analysis time and no need of standard compounds for calibration curves preparation. These findings indicate that this method has potential as a reliable method for quality evaluation of herb medicines, especially for protoberberine alkaloid-containing ones.

[Establishment of seedling classification criteria of Coptis chinensis by dynamic clustering method].

OBJECTIVE: To establish seedling classification criteria of Coptis chinensis. METHOD: The height of plant, leaf number, leaf length, leaf wide, weight of leaf, weight of root were measured, the main measurement indexes of seedlings of C. chinensis were chosen through correlation and regression analysis. The seedling classification criteria were formulated by dynamic clustering analysis. RESULT: The criteria of the 1st-grade seedlings were as follows: leaf number above 8 leaves, height of plant between 12 and 14 cm. The criteria of the 2nd-grade seedlings were as follows: leaf number between 6 and 8 cm, height of plant between 9 and 12 cm. The criteria of the 3rd-grade seedlings were as follows: leaf number between 4 and 6 cm, height of plant above 9 cm. CONCLUSION: The seedling classification criteria of C. chinensis was scientific and feasible, and can be used for the quality control standard of C. chinensis.

Metabolic discrimination of rhizoma coptidis from different species using 1H NMR spectroscopy and principal component analysis.

Rhizoma coptidis, a broadly used medicinal plant, originates from the dried rhizomes of three species in Chinese pharmacopoeia, namely, Coptis chinensis Franch, Coptis deltoidea C. Y. Cheng et Hsiao, and Coptis teeta Wall. In this study, a novel approach using (1)H NMR spectroscopy combined with multivariate analysis was introduced to differentiate the three species and identify potential metabolic markers for better controlling the quality of rhizoma coptidis. A broad range of metabolites including alkaloids, sugars, organic acids, amino acids, and fatty acids present in rhizoma coptidis were detected by means of (1)H NMR spectroscopy. Principal component analysis (PCA) of the (1)H NMR data set showed a clear separation between all samples by PC1 and PC3, and some metabolites that could be responsible for the discrimination of the three species were identified. An analysis of variance (ANOVA) was performed to statistically verify the significance of differences in metabolite levels between species. By combining PCA and ANOVA, significantly higher contents of palmatine, coptisine, epiberberine, columbamine, and fatty acids together with lower contents of jateorrhizine were found in Coptis chinensis, whereas Coptis deltoidea and Coptis teetA showed the highest levels of sucrose and chlorogenic acid, respectively. This study indicates that metabolites of rhizoma coptidis vary with the species and the proposed method is suitable for metabolic fingerprinting analysis to check the genuine origin of rhizoma coptidis.

[Classification study of Coptis chinensis based on quantitative physical property characteristics of appearance and internal quality evaluation].

OBJECTIVE: Combining the quantitative physical property characteristics of the appearance with the internal quality evaluation index, its aims to provide experimental basis for the classification and quality evaluation of Coptis chinensis. METHOD: Fourteen batches of C. chinensis from different areas were respectively measured in size (total length, total width, root length, taproot diameter, branch number, branch length, branch diameter, length of the bridge, weight), color (external color, internal color), content (epiberberine, coptisine, palmatine, berberine). Then the determination data were evaluated by spss principal component analysis and cluster analysis. RESULT: Three principal components were extracted from the original data. The principal component analysis results showed that the characteristic elements might be the total length, main root length, taproot diameter, branch length, weight, the total color value of the appearance and content of epiberberine and berberine. The results of cluster analysis showed that 14 batches of samples could be clustered reasonably into two groups. In terms of the appearance and quality, there were some differences between in the geo-authentic and non-authentic producing areas of C. chinensis. CONCLUSION: The method which was combining the quantitative physical property characteristics of the appearance with the internal quality evaluation index, and through the processing of mathematical statistics, could be used for the the classification of C. chinensis.

[Study on the net photosynthesis rate of Coptis chinensis from different types and production places].

OBJECTIVE: Evaluate the photosynthesis characteristics of Coptis chinensis and provide theoretical basis for the optimization of Coptis chinensis germplasm resources and variety. METHODS: The daily variation of the net photosynthetic rate, transpiration rate, stomatal conductance and intercellular CO2 density in leaf blade were determined by Photosynthesis analyzer, and the net photosynthesis rate of Coptis chinensis from different plant ages, types and production places was compared. RESULTS: The light saturation point in Coptis chinensis was about 500 micromol/m2 x s, light compensation point was about 12.04 micromol/(m2 x s), apparent quantum yield was 0.011. When the temperature was above 33 degrees C, the transpiration rate increased but the net photosynthesis rate decreased. There was no significant difference (P > 0.05) among the net photosynthesis rate of Coptis chinensis from different plant ages. The net photosynthesis rate of the type DA-YE and ZHI-HUA was significantly higher than that of other types (P < 0.05); The net photosynthesis rates of specimens from Fubao mountain in Lichuan county of Hubei and Fengmu village in Shizhu county of Chongqing were significantly higher than that of others. CONCLUSIONS: The net photosynthesis rate of Coptis chinensis decreased under hight temperation and hight light. Different germplasm resources of Coptis chinensis, the net photosynthetic rate had certain differences.

[Herbalism and germplasm of deltaleaf goldthread rhizome].

Through textual research of herbalism about Chinese Goldthread Rhizome in different periods of ancients and some modern study, and analysis on correlativity between its name and different germplasm, it showed that the knowledge level of quality of the medicinal materials was gradually increased in its historical record of application by the society improvement and increasing of requirements and population from producing area, diversity of character to germplasm resources. The materials name of Chinese Goldthread Rhizome was from unitary to diversity and referred to the rhizomes of all plants of Coptis in China, while Deltaleaf Goldthread Rhizome took an important role in the evolution of Chinese Goldthread Rhizome. The aim of the work was to provide a reference for the correlative study of phylogeny, individual distribution, ecological environment and medical resources.

[RAPD analysis for genetic diversity of medicinal plant Coptis omeiensis].

OBJECTIVE: To discuss the genetic diversity of Coptis omeiensis. METHOD: The genetic diversity of 110 individuals from 10 populations was analyzed by RAPD. RESULT: 14 primers were selected to produce highly reproducible RAPD bands. Among 132 amplified bands, 98 showed polymorphism, the percentage of polymorphic bands reached to 74.24%. Nei's gene diversity index (H) was 0.2863, Shannon's information index (I) was 0.3624, G(st) was 0.2305. The genetic distance coefficient and the similarity were 0.1931-0.5245 and 0.5016-0.8843, respectively. CONCLUSION: There exists a held high genetic diversity in C. omeiensis and the majority of genetic variation occurs in the populations. By cluster analysis, the geographical distribution is very obvious. The RAPD marker can be used for the analysis of the genetic diversity and genetic variation of C. omeiensis.

[ISSR analysis of genetic diversity of Coptis deltoidea].

OBJECTIVE: To study the genetic diversity of Coptis deltoidea. METHOD: The genetic diversity of 90 individuals from 8 populations was analyzed by ISSR. RESULT: Twelve primers were selected to produce highly reproducible ISSR bands. Among 128 amplified bands, 94 showed polymorphism, the percentage of polymorphic bands reached 73.44%. Nei's gene diversity index (H) was 0.1925, Shannon's information index (I) was 0.3028, Gst was 0.7212. The genetic distance coefficient and similarity were 0.0858-0.2314 and 0.8046-0.9425, respectively. CONCLUSION: C. deltoidea held a high genetic diversity and the majority of genetic variation occurs among populations. By cluster analysis, the geographical distribution is very obvious. The ISSR marker can be used for the analysis of genetic diversity and genetic variation of C. deltoidea.

Galactinol synthase gene of Coptis japonica is involved in berberine tolerance.

Many plant secondary metabolites show strong biological activities and are potentially also toxic to plants, while plants producing such active compounds are usually insensitive to their own metabolites, suggesting that they have species-specific detoxification mechanisms. In order to clarify the detoxification mechanism of alkaloids, we used cultured cells of Coptis japonica, which are capable of producing a yellow benzylisoquinoline alkaloid, berberine, and accumulate it in the vacuole. Unlike other plant cells that do not produce berberine, C. japonica shows strong tolerance to this alkaloid. We established a fission yeast strain that was sensitive to berberine and performed functional screening using a C. japonica cDNA library. One cDNA clone, which conferred clear berberine tolerance, encoded galactinol synthase (CjGolS). The possible role of CjGolS in berberine tolerance is discussed.

[Fingerprints of Rhizoma Coptidis from Shizhu by HPLC].

OBJECTIVE: To establish a sensitive and specific HPLC method for the quality control of Rhizoma Coptidis collected from shizhu in chongqing. METHODS: The HPLC fingerprints of Rhizoma Coptidis from shizhu were obtained from Waters instrument. The methods was performed on a Diamonsil C18 column gradient eluted with acetonitrile-0.05 mol/L KH2PO4 (pH3 with H3PO4) at the flow rate of 0.8 m/min. The temperature of column was 25 degrees C and the UV detection wavelength was 270nm. RESULTS: The HPLC fingerprint of Rhizoma Coptis, showing 16 characteristic peaks, was established from 10 lots of Rhizoma Coptis. By comparision of the retention time and the on-line UV spectra of chemical standards, peak 11, 12, 13, 14 and 15 were identified as Epiberberine, Jatrorrhizine, Coptisine, Palmatine and Berberine. CONCLUSION: The HPLC fingerprint of Rhizoma Coptidis with high specificity can be used to control its quality.

[Genetic diversity of Coptis chinensis germplasm based on ISSR analysis].

OBJECTIVE: To detemine the genetic diversity of Coptis chinensis. METHOD: 32 germplasmic resources of C. chinensis were analyzed by ISSR molecular markers. To make up the systematic diagram of genetic relationship by TREECONW software and clustered by UPGMA method. RESULT: A total of 106 ISSR bands were scored, among which 51 were polymorphic bands. The average percentage of polymorphic bands was 48. 1%. Genetic similarity coefficient were changed from 0. 825 8 to 0. 935 1. By cluster analysis, the geographical distribution is not very obvious, but it was also showed some of the C. chinensis from the same region were in the same group which presented the law of geographical distribution in the tested materials. CONCLUSION: Different germplasms diversity of C. chinensis is low and the relationship of C. chinensis is close.

[Population characteristics of Coptis chinensis var. brevisepala in Anhui Province and its endangering mechanism].

The study on the distribution pattern, population size, and age structure of C. chinensis var. brevisepala in Anhui Province showed that C. chinensis var. brevisepala mainly distributed in the southern mountain areas such as Mts. Jiuhuashan, Guniujiang, Huangshan and Qingliang Peak. The habitats had two kinds, i.e., under woods and beside water ditches, both of which were shady and humid, with loose and acidic soils containing abundant organic matter. The shading degree surpassed 70%, air humidity reached 70% approximately 90%, and soil water content was more than 30%. The distribution pattern of existing C. chinensis var. brevisepala populations was clustering, with illogical age structure and small population size. It was suggested that the populations of C. chinensis var. brevisepala in Anhui Province were in an unsteady stage. Serious destroy of natural habitat and excessive digging and gathering adult individuals were the main causes of the rapid decrease of the individuals in some populations, which threatened the existence of C. chinensis var. brevisepala populations seriously.

[Quality assessment of Coptis chinensis in China].

OBJECTIVE: To study the dynamic change of the ingredients of Coptis chinensis and evaluate the quality of the crude drugs from main producing areas. METHOD: The ingredients of samples from Shizhu and Wuxi counties in Chongqing, Hongya and Dayi counties in Sichuan, Zhenping county in Shanxi, Lichuan county in Hubei were analyzed for berberine, coptisine, jatrorrhizine, palmatine and the total alkaloids by HPLC and UV methods. RESULT AND CONCLUSION: The contents of measured indexes were mostly highest in 5-years-old C. chinensis. Considering factors such as the yield, it is reasonable to harvest the 5-years-old C. chinensis. There are minor differences in condents of C. chinensis from different areas, all tested samples met the pharmacopoeial standards. It is concluded that the general quality of C. chinensis was good and acceptable.

[Research development of Coptis chinensis].

To summarize the research and application of ecological planting technique for Coptis chinensis, and describe the recent development of its chemical components, pharmacological effects and clinical applications.