Study on curcumins from Curcuma phaeocaulis / 中草药

Objective: To study the chemical constituents and the cytotoxic activity of curcumins from the rhizome of Curcuma phaeocaulis. Methods: The 95% ethanol extract from the rhizome of C. phaeocaulis was extracted with petroleum ether, ethyl acetate, and n-butanol. The separation and purification of ethyl acetate fraction was carried out by silica gel column, sephadex LH-20 column, reversed-phase medium pressure chromatography, preparative thin-layer chromatography, and semi-preparative high performance liquid chromatography. The structures of the isolated components were identified by modern spectroscopy techniques. The isolated compounds were screened for cytotoxic activity by MTT assay. Results: Four curcuminoids were isolated from the ethyl acetate extract of the rhizome of C. phaeocaulis, and identified as 1,7-bis (4-hydroxyphenyl)-1E,6E-heptadien-3-one (1), 1,7-bis (4-hydroxyphenyl)-1,4,6-heptatrien-3-one (2), 1,7-bis (4-hydroxyphenyl)-4E,6E-heptadien-3-one (3), and (1R,5S,6S)-1,5-epoxy-6- hydroxy-1,7-bis (3-methoxy-4-hydroxy-phenyl)-heptane (4). MTT experiments showed that compounds 1-3 inhibited HGC-27 cells proliferation, and only compound 2 inhibited MDA-MB-231 cells proliferation. Compounds 2 and 3 also showed strong toxic effects on human normal liver cells. Conclusion: Four curcuminoids were isolated from C. phaeocaulis. Compound 1 was a new compound named curcumin P. Compounds 1-3 had a certain inhibitory effect on the proliferation of HGC-27 cells. Notably, compound 1 selectively inhibited the proliferation of HGC-27 cells and showed no obvious toxic effects on L-02 cells.

Qualitative and quantitative analysis of trace element of terpenoid conjugated curcuminoid and curcuminoids in herbal medicine derived from different Curcuma species / 中草药

Objective: To qualitatively and quantitatively analyze the terpene-conjugated curcuminoid compound- Bisabolocurcumin ether (BCE) in Curcuma species. Methods: UPLC-Q-TOF-MS/MS was used to establish a qualitative analysis of BCE, and its distribution in Curcuma species was studied. UPLC was used to establish quantitative analysis of BCE and three main curcuminoids in three Curcuma species (Rhizoma Curcumae Longae, HuangsiYujin, Curcuma phaeocaulis), in order to study the difference of BCE content in different Curcuma species and its content difference with other curcuminoids. Results: In the qualitative analysis, by analyzing the primary, secondary pyrolysis fragments and the cracking law of BCE, it was found that only Jianghuang and HuangsiYujin contained it, but BCE could not be detected in the PengEzhu. In the quantitative analysis, BCE and the three main curcuminoids had good linear relationship in their linear range (r ≥ 0.999 3); The average recovery rates were in range of 99.407%-113.631%, and RSD < 4%. It was found that the content of three curcuminoids in Jianghuang were significantly higher than the other two herbs, and the content of BCE in the three herbs was very different. The BCE content in Jianghuang was four times than that of HuangsiYujin. Conclusion: The qualitative and quantitative analysis methods established in this study can provide reference for the discovery and analysis of more novel BCE structure’s compounds, and provide a basis for improving the quality standards of Curcuma species.

Comparation of intestine absorption kinetics of volatile oil of Sichuan genuine herb Curcuma phaeocaulis before and after vinegar processing in rats / 中草药

Objective: To Compare the difference of the intestinal absorption kinetics of volatile oil of Sichuan genuine herb Curcuma phaeocaulis before and after vinegar processing. Methods: Rat model of systemic intestinal perfusion were established and were divided into four groups including volatile oil of C. phaeocaulis group, volatile oil of C. phaeocaulis processed with vinegar group, volatile oil of C. phaeocaulis added with curcuminoids group and volatile oil of C. phaeocaulis processed with vinegar added with curcuminoids group to investigate the absorption of effective constituents of germacrone and furanodiene in the intestinal segements, and calculate the absorption rate constant (Ka), absorption half-life (t1/2), and absorption rate per unit time (P). Results: The results showed that the absorption degree and Ka value of volatile oil of C. phaeocaulis after vinegar processing was less than that before vinegar processing. The degree and Ka value of volatile oil after addition of curcuminoids was less than that before compatibility. Conclusion: C. phaeocaulis after processing with vinegar could alleviate drug property through delaying the absorption of volatile oil to stabilize the blood concentration, with view to meeting the need of clinical safety ad effectivity.

Protein network module-based identification of key pharmacological pathways of Curcuma phaeocaulis Val. acting on hepatitis.

ETHNOPHARMACOLOGICAL RELEVANCE: Curcuma phaeocaulis Val. (CP), as the vital medicines for blood-breaking and disorder-eliminating, has been widely used for hepatitis with good curative effects. Owing to the complexity of traditional Chinese medicine, the pharmacological mechanism of CP remains unclear. To solve this problem, a protein network module-based approach was proposed in this study. MATERIALS AND METHODS: Firstly, the content of active components of CP was detected based on HPLC-DAD. Then the liver protection of CP on Con A-induced hepatitis was validated via the analysis of serum levels of ALT, AST and LDH and histological findings. Next, the targets of CP components obtained from TCMD database were predicted by STITCH and ChEMBL retrieval. In addition, the protein interaction network (PIN) of CP was constructed by Cytoscape based on protein-protein interaction of targets obtained from STRING database. Following the topological analysis of CP PIN, it showed to exhibit the properties of scale-free, small world, and modularity matched with the property of complex biological networks. Finally, the functional modules were identified by gene ontology enrichment analysis based on Molecular Complex Detection algorithm. RESULTS: The functional modules indicated that the mechanism of CP acting on the hepatitis is significantly associated with NF-κB and TGF-ß signaling pathway. More interestingly, curcumin, demethoxycurcumin and bisdemethoxycurcumin were the main active components of CP acting on the hepatitis, which were demonstrated to be associated with the inflammatory process that occurs during the progression of hepatitis. CONCLUSION: The protein network module-based approach is an efficient way to investigate the pharmacological mechanisms of CP.

Research on changes of chemical components in Sichuan authentic herbs of Curcuma Phaeocaulis processed with rice vinegar and decocted by detecting based on dynamic variation of pH value / 中草药

Objective: To determine the dynamic variation of pH value and seven components including bisdemethoxycurcumin, demethoxycurcumin, curcumin, curdione, curcumol, germacrone and β-elemene during Curcuma Phaeocaulis' processing, in order to study its curcuminoids and volatile oils components in the process of rice vinegar production affected by external factors such as water, heat, and acid. Methods: C. phaeocaulis were processed by using distilled water, 9° rice vinegar, and 9% acetic acid aqueous. The pH value of the decoction at a specific time point in 10 batches was detected. The data were analyzed by SPSS software. The content of seven components in the samples of the raw crude group, the blank group, the vinegar group, and the reference group were determined by HPLC method, and the change of the content was analyzed. Results: Compared with the blank group, the vinegar group and reference group showed low pH value, and there was no significant difference; Compared with the blank group and reference group, the contents of three curcuminoids in the vinegar group increased [the mass fraction of bisdemethoxycurcumin is (0.002 320 ± 0.000 344) mg/g, the mass fraction of demethoxycurcumin is (0.059 65 ± 0.015 64) mg/g, the mass fraction of curcumin is (0.272 5 ± 0.125 2) mg/g], and the contents of volatile oils decreased in different degrees. Conclusion: During the processing progress, the main active constituents of pharmaceutic adjuvant 9° rice vinegar might be acetic acid; The acetic acid and other organic principles of rice vinegar might protect the chemical constituents in C. phaeocaulis by regulating pH value; The toxicity degree of C. phaeocaulis before and after processing with vinegar was regulated by different protecting degrees of different component. It not only reflects the scientificity of 9° rice vinegar as a pharmaceutic adjuvant, but also offers a new method to study the material basis of C. phaeocaulis processing with rice vinegar.

Anti-inflammatory, antioxidant and antitumor activities of ingredients of Curcuma phaeocaulis Val.

Curcuma phaeocaulis Val. is used in Chinese Pharmacopoeia as health food and folk medicine for removing blood stasis, alleviating pain and tumor therapy. This research was aimed to explore and compare three main bioactivities including anti-oxidant, antitumor and anti-inflammatory activities between the ethanol extract of C. Phaeocaulis and its fractions using different in vitro models. Firstly, 70 % ethanol was used to extract C. Phaeocaulis, and then the crude extract was re-extracted, resulting in petroleum ether (EZ-PE), ethyl acetate (EZ-EA), and water fractions (EZ-W), respectively, and then a series of index was detected. Results showed that all the extracts had medium DPPH radical scavenging activity when the concentration was 200 µg/ml and their DPPH radical scavenging activity was in a concentration-dependent manner. The extracts except ethanol extract of C. Phaeocaulis had almost no cytotoxicity to the survival of RAW264.7 cell when the concentration reached 80 µg/ml, and all of them had medium inhibitory effect on nitrite release. Extracts of C. Phaeocaulis had medium intensity antitumor activity, EZ-PE and EZ-EA fractions significantly inhibited the proliferation of four tumor cells (SMMC-7721 cell lines, HepG-2 cell lines, A549 cell lines and Hela cell lines). C. Phaeocaulis had antioxidant and anti-inflammatory activities, which did not carry out centralized phenomenon when re-extracted. EZ-PE and EZ-EA were active antitumor sites of C. Phaeocaulis.

Pharmacokinetic studies on curcumin in Curcuma phaeocaulis in rats in vivo / 中草药

Objective To study pharmacokinetics of curcumin in Curcuma phaeocaulis in rats in vivo.Methods HPLC method was used to determine the curcumin in rat plasma.The conditions were column: Lichrospher-5-C_(18)(250 mm?4.6 mm, 5 ?m); column temperature: 25 ℃;mobile phase: CH_3CN-5% HAc water solution(45∶55);flow: 1 mL/min;detection wavelength: 420 nm.Results The calibration curve was liner(r=0.999 5) at the range of 6.5—104 ?g/mL.The average recovery was 98.5%.RSD was 2.41%(n=5).The pharmacokinetic parameters of curcumin were as follows: k_a was 0.53/h,k_e was 0.10/h,t_(1/2ka) was 1.32 h,t_(1/2k) was 6.89 h,t_(peak) was 3.89 h,C_(max) was 93.15 ng/mL,AUC was(1 369.38) ng/mL.Conclusion This method is stable,simple,and reliable,which can be applied for the determination of curcumin in plasma and pharmacokinetic studies.