ABSTRACT
Objective To obtain the transcriptome sequence database and to identify the genes related to the biosynthesis of anthraquinones in Rubia cordifolia. Methods The transcriptome data of root of Rubia cordifolia was obtained by Illunima HiseqTM 2000 150PE sequencing and de novo splicing of Unigene was realized by Trinity. The GO classification, KOG functional, metabolism of KEGG metabolic pathway and protein function annotation analysis were completed based on BLAST. Results 7.3 Gb database was assembled after assembly steps, and 66 646 unigenes were assembled with an average length of 1 424 bp. All Unigenes were annotated in the public databases NR (NCBI nonredundant protein sequence), NT (NCBI nucleotide sequences), KEGG (kyoto encyclopedia of genes and genomes), Swissprot (A manually annotated and reviewed protein sequence database), GO (gene ontology), KOG (euKaryotic ortholog groups) and Pfam (protein family). Based on the assignment of KEGG pathway, 64 Unigenes involved in anthraquinones biosynthesis were found. Conclusion This study was the first comprehensive transcriptome analysis for R. cordifolia, and the candidate genes involved in the biosynthesis of anthraquinones were obtained. The transcriptome data constitutes a much more abundant genetic resource that can be utilized to benefit further molecular biology studies on R. cordifolia.
ABSTRACT
Objective To evaluate the effects of a traditional Chinese medicine Rubia cordifolia L.aqueous extract (RCAE) on body weight, fat mass and parameters of glucose and lipid metabolism in high fat diet (HFD)-induced obese rats and its mechanism.Methods pGL3-Enhancer-PPARγ2 (625 bp)-Luc plasmid, a luciferase reportergene expression plasmid containing PPARγ2 promoter was constructed and stably transfected 3T3-L1 preadipocytes were established.PPARγ2 promoter`s activities in these cells were detected after administration with different concentration (0.1 mg/L~1 000 mg/L) of RCAE or with 100 mg/L RCAE for different action time.PPARγ2 mRNA expression in human adipocytes were detected after administration with 100 mg/L RCAE.Meanwhile, HFD-induced obese rats were administrated with low or high dose RCAE to investigate the effects of RCAE on serum glucose, lipid and insulin levels, body weight, visceral fat mass and so on.Results 10 mg/L RCAE could increase luciferase expression in 3T3-L1 cells to 1.43 folds of that in control group (P<0.01) and it reached 3.24 folds of that in control group when the concentration of RCAE was 1000 mg/L (P<0.01).With the administration with 100 mg/L RCAE, the luciferase activity of 3T3-L1 cells peaked at 28 h where it was 2.72 folds of that in control group (P<0.01), and the expression of PPARγ2 mRNA in human adipocytes increased to 2.27 folds of that in control group (P<0.01).Compared with HFD group, low dose RCAE significantly reduced the fasting insulin level, HOMA-IR and visceral fat mass (P<0.05).Conclusions Low dose RCAE significantly reduces the visceral fat mass and ameliorates insulin resistance in HFD-induced obese rats.The potential mechanism may be explained by the stimulation of PPARγ2 promoter activities and the increased expression of PPARγ2 gene.
ABSTRACT
Objective To investigate the chemical constituents of the aerial part of Rubia cordifolia. Methods All compounds were isolated and purified by silica gel, Sephadex LH-20, and MCI column chromatography. Their structures were determined by physicochemical properties and spectral data. Results Eleven compounds were isolated and identified as: (-)-episyringaresinol (1), (-)-secoisolariciresinol (2), (-)-3,4-divanillyl tetrahydrofuran (3), lignans (+)-demethoxypinoresinol (4), (+)-syringaresinol (5), (+)-isolariciresinol (6), burselignan (7), (7S,8R)-dihydrodehydroconiferyl alcohol (8), 4,5'-dimethoxy-lariciresinol (9), (+)-7R,8S-5- methoxydihydrodehydroconifery alcohol (10), and 5,5'-dimethoxy-7-oxolariciresinol (11). Conclusion All compounds are isolated from this plant for the first time.
ABSTRACT
Rubia cordifolia L. has been used as a traditional Chinese medicine for several centuries. In recent years, the resources of wild Rubia cordifolia have been declined sharply due to increased utilization and rising price. Therefore, it is of great urgency to evaluate and protect resources of wild plant of Rubia cordifolia. In our study, sixty-four individuals that represent eight wild populations of Rubia cordifolia L. were analyzed by Start Codon Targeted Polymorphism (SCoT) molecular markers. Genetic distance was calculated by POPGENE 3.2 software, cluster analysis was generated by NTSYS 2.10 software based on UPGMA method and Mantel Test was used to analysis the relationship between the genetic distances and geographical distance among the wild populations. The results showed a high genetic diversity of wild populations of Rubia cordifolia L. in Shaanxi province. A total of 182 bands were produced by 14 primers, among which 163 bands were polymorphic bands, and the percentage of polymorphic bands was 89.56%. The average value of Nei's genetic diversity index (H) was 0.293 6, Shannon's information index (I) was 0.444 6, genetic differentiation coefficient (Gst) was 0.555 3, and the gene flow (Nm) was 0.440 8, the wild populations were ranked by genetic diversity:AK > YL > SL > BJ > TC > YA > WN > XY. Mantel Test analysis demonstrated that the significant correlation was found between the genetic distances and geographical distances (r=0.776 4, PRubia cordifolia L. germplasms.
ABSTRACT
To prepare the intrauterine slow release silicone rubber bar made of Panax notoginseng and Rubia cordifolia, and finish its preliminary evaluation of in vitro releasing properties. The open mill method was used for plasticating of silicone rubber. The process parameters of the silicone rubber and drugs mixing were optimized by orthogonal test. The parameters of silicone rubber vulcanization was optimized by single factor test. The preliminary evaluation of in vitro release performance of the silicone rubber bar was conducted with ginsenoside Rg₁, ginsenoside Rb₁, notoginsenoside R₁, purpurin and rubimaillin as the indexes. The results showed that optimum technologic parameters for silicone rubber and drugs mixing：the roller spacing 2 mm; speed ratio 1∶1.2; front roller temperature 55-60 ℃; rear roll temperature 50-55 ℃; and mixing time 20 min. The optimum parameters for silicone rubber vulcanization：temperature 90 ℃, and time 60 min. The studies on release process in vitro revealed that the release process of silicone rubber bar was in line with the Higuchi equations. After 90 days, the cumulative release of ginsenoside Rg₁, ginsenoside Rb₁ and notoginsenoside R₁ was 46.7%, and the cumulative release of purpurin and rubimaillin was 51.9%. The preparation method can be applied to the preparation of silicone rubber bar, with slow release characteristics.
ABSTRACT
OBJECTIVE: To identify traditional Chinese medicine Rubiae Radix et Rhizoma and its adulterants using DNA barcodes. METHODS: A total of 317 sequences of psbA-trnH, matK and rbcL from 13 species were analized. The intra- and inter-specific K2P genetic distances and neighbor-joining tree were calculated using MEGA5. 1 program. RESULTS: The DNAs were successfully extracted from 76 original plant samples. The amplification rates of psbA-trnH, matK and rbcL were 100%, 96%, and 99%, respectively. The adulterants could be identified from R. cordifolia by the single marker and the two combinations except for those adulterants in the Rubia genus. Fortunately, R. cordifolia could be identified from all the adulterants by psbA-trnH + matK + rbcL combination marker. The intra- and inter-specific K2P genetic distances of psbA-trnH + matK + rbcL were 0-0.001 4 and 0.000 7-0.630 3. R. cordifolia could be identified from alTthe adulterants by the psbA-trnH + matK + rbcL combination using NJ tree method. Among the 30 unidentified samples of Rubiae Radix et Rhizoma which were collected from medicinal herb markets, all three markers, including psbA-trnH, matK and rbcL, could be amplified from 22 samples. The 11 samples of Rubiae Radix et Rhizoma clustered with R. cordifolia and the others were divided into three clusters by the psbA-trnH + matK + rbcL combination using NJ tree method. CONCLUSION: psbA-trnH + matK + rbcL combination can be used as DNA barcode to identify R. cordifolia from adulterants.
ABSTRACT
Objective: To investigate the genetic diversity of Rubia cordifolia from seven geographical populations in Henan province, and to provide evidence for the protection of R. cordifolia wild resources. Methods: The autogenous variation of chloroplast gene fragment psbA-trnH was analyzed by DNA sequencing technique. Results: Based on cpDNA data, 40 haplotypes (H1-H40) were identified, the diversity index of haplotype and nucleotide diversity index were 0.951 ± 0.0013 and 0.01477, respectively. Genetic differentiation index was 0.08153, and gene flow among populations was 2.78. Conclusion: The wild populations of R. cordifolia in Henan province have higher genetic diversity in geo-authentic product area than other species. There is less genetic differentiation among populations because of the stronger seed-mediated gene flow.
ABSTRACT
AIM@#To determine the IPP origin of the naphthoquinones (NQs) in Rubia cordifolia, and to evaluate the effects of methyl jasmonate (MeJA) treatment, MEP, and MVA pathway inhibitor treatment on the accumulation of anthraquinones (AQs) and NQs in cell suspension cultures of R. cordifolia.@*METHODS@#Cell suspension cultures of R. cordifolia were established. Specific inhibitors (lovastatin and clomazone) and MeJA were supplied to the media, respectively. Treated cells were sampled every three days. Content determination of purpurin (AQs) and mollugin (NQs) were carried out using RP-HPLC. The yield of the two compounds was compared with the DMSO-supplied group and the possible mechanism was discussed.@*RESULTS@#Lovastatin treatment increased the yield of purpurin and mollugin significantly. Clomazone treatment resulted in a remarkable decrease of both compounds. In the MeJA-treated cells, the purpurin yield increased, meanwhile, the mollugin yield decreased compared with control.@*CONCLUSION@#The IPP origin of mollugin in R. cordifolia cell suspension cultures was likely from the MEP pathway. To explain the different effects of MeJA on AQs and NQs accumulation, studies on the regulation and expression of the genes, especially after prenylation of 1,4-dihydroxy-2-naphthoic acid should be conducted.
Subject(s)
Acetates , Pharmacology , Anthraquinones , Metabolism , Cell Culture Techniques , Cells, Cultured , Cyclopentanes , Pharmacology , Isoxazoles , Pharmacology , Lovastatin , Pharmacology , Oxazolidinones , Pharmacology , Oxylipins , Pharmacology , Pyrans , Metabolism , Rubia , MetabolismABSTRACT
To evaluate the in vivo antioxidant activity of alcoholic extract of the roots of Rubia cordifolia (RC). Male mice, were exposed, either to lead nitrate at a dose containing 40mg/kg body weight or combined with Rubia cordifolia for the first period of 40 days, where the animals were sacrificed for oxidative studies and biochemical studies. Lead has induced a significant increase in LPO, where as significant depletion SOD, CAT and GSH in liver and testis tissues. Rubia cordifolia alone had moderate effect on mice, where as coadministration of lead nitrate with Rubia cordifolia reversed the effect of lead. Ingestion of Pb (NO3)2 showed significant elevation in AST, ALT, ACP, ALP, and total cholesterol, level in tissue homogenate whereas Total protein content decreased significantly in comparison to control animals. On the other hand Alcoholic extract of Rubia cordifolia (low and high dose), along with lead nitrate decreased, elevated levels of AST, ALT, ACP, ALP, and total cholesterol, as compared to lead nitrate intoxicated mice. A significant rise in the level of total protein was also noticed. Preliminary analysis has revealed that Rubia cordifolia has significant amount of GSH, Vitamin C, other important antioxidants and polyphenols. In addition it also contains important trace elements like Zn, Cu, Vd, Se and Mo. These contribute to its antioxidant properties.
ABSTRACT
Objective To discuss the mechanism of rubia cordifolia extractive affecting some biochemical indicators of blood serum and sporting ability in highly endurance-trained rats,which can provide theoretical basis for using rubia cordifolia extractive as sport supplements.Methods After establishing the highly endurance-trained rats model,we tested the exhaustion time in the trained rats and the contents of serum urea(BUN),lactic dehydrogenase(LOH),blood lactate(BLa)as well as the cruarin(Hb)in the rats.Results The activity of LDH in training group was greater than that in silent group(P