Tectoridin Stimulates the Activity of Human Dermal Papilla Cells and Promotes Hair Shaft Elongation in Mouse Vibrissae Hair Follicle Culture.

To search hair growth-promoting herbal extract, a screening platform of having HEK293T fibroblast being transfected with pTOPFLASH DNA construct was developed over a thousand of herbal extracts and phytochemicals were screened. One of the hits was ethanolic extract of Rhizoma Belamcandae, the rhizome of Belamcanda chinensis (L.) DC. Tectoridin, an isoflavone from Rhizoma Belamcandae, was shown to be responsible for this activation of promoter construct, inducing the transcription of pTOPFLASH in the transfected fibroblasts in a dose-dependent manner. The blockage by DKK-1 suggested the action of tectoridin could be mediated by the Wnt receptor. The hair growth-promoting effects of tectoridin were illustrated in human follicular dermal papilla cells and mouse vibrissae organ cultures. In tectoridin-treated dermal papilla cultures, an activation of Wnt signaling was demonstrated by various indicative markers, including TCF/LEF1 transcriptional activity, nuclear translocation of ß-catenin, expressions level of mRNAs encoding axin-related protein, (AXIN2), ß-catenin, lymphoid enhancer-binding factor-1 (LEF-1), insulin-like growth factor 1 (IGF-1) and alkaline phosphatase (ALP). In addition, an increase of hair shaft elongation was observed in cultured mouse vibrissae upon the treatment of tectoridin. Tectoridin, as well as the herbal extract of Rhizoma Belamcandae, possesses hair promoting activity, which deserves further development.

Efficient degradation and enhanced anticomplementary activity of Belamcanda chinensis (L.) DC. polysaccharides via trifluoroacetic acid treatment with different degrees.

The degradation of Belamcanda chinensis (L.) DC. polysaccharides was carried out by five concentrations of trifluoroacetic acid (TFA) (1-5 mol/L), and their physicochemical properties, degradation kinetics and anticomplementary activity were investigated. The findings revealed a notable reduction in the molecular weight of BCP, from an initial value of 2.622 × 105 g/mol to a final value of 6.255 × 104 g/mol, and the water solubility index increased from 90.66 ± 0.42 % to 97.78 ± 0.43 %. The degraded polysaccharides of B. chinensis exhibited a comparable monosaccharide composition comprising Man, GalA, Glc, Gal, and Ara. As the concentration of TFA increased, the degradation rate constant increased from 1.468 × 10-3 to 5.943 × 10-3, and the process followed the first-order degradation kinetic model (R2 > 0.97) and the random fracture model (R2 > 0.96). Furthermore, the five degraded polysaccharides still exhibit good thermal stability. In vitro experiments showed that DBCP-3 exhibited more potent anticomplementary activity than the original polysaccharides and positive drugs, which was strongly correlated with its Mw (r = 0.6-0.8), inhibiting complement activation by blocking C2 and C4. These results indicated that TFA degradation has a positive effect on polysaccharides, of which DBCP-3 is expected to treat diseases involving hyperactivation of the complement system.

A novel multidimensional strategy to evaluate Belamcanda chinensis (L) DC and Iris tectorum Maxim based on plant metabolomics, digital reference standard analyzer and biological activities evaluation.

BACKGROUND: Belamcanda chinensis (L.) DC. (BC) belongs to the family of Iridaceae and is widely cultivated and used in many Chinese patent medicine and Chinese medicinal formulae. However, due to the high similarities in appearance such as color and shape to Iris tectorum Maxim (ITM), another plant from the same family, BC is often confused or even misused with ITM. METHODS: Therefore, in order to distinguish the chemical constituents, qualities and biological activities of BC and ITM, multiple technologies including plant metabolomics, digital reference standard (DRS) analyzer and biological activities assay were employed to provide a sufficient basis for their practical applications. RESULTS: In plant metabolomics, the PCA and OPLS-DA score plot indicated the obvious differences in chemical profiling between BC and ITM and 6 compounds were successfully identified to contribute to the differences. In DRS study, the fingerprints of 10 and 8 compounds in BC and ITM were developed based on DRS analyzer, respectively, involving relative retention time (RRT) method and linear calibration using two reference substances (LCTRS) technique. The DRS analyzer also accurately identified 10 and 8 compounds from BC and ITM, respectively, by using only two reference standards. In biological activities assay, BC had a better anticancer effect than ITM due to the high abundance of irigenin, while ITM showed stronger hepatoprotective activity than BC because of the high abundance of tectoridin. CONCLUSIONS: Therefore, due to the significant differences of B. chinensis and I. dichotoma in chemical composition and biological activities, the current studies strongly proved that these two medicinal plants could not be mixed in industrial production and clinical medication.

Tissue-specific metabolites profiling and quantitative analyses of flavonoids in the rhizome of Belamcanda chinensis by combining laser-microdissection with UHPLC-Q/TOF-MS and UHPLC-QqQ-MS.

The rhizome of Belamcanda chinensis (L.) DC. is a traditionally used medicinal material in China. Due to increasing demand, B. chinensis has been cultivated widely, and thus the study on its rational utilization of medicinal part and guidelines for the optimal cultivation and harvest is an important issue. Considering flavonoids were the main bioactive secondary metabolites of B. chinensis, fluorescence microscopy, laser microdissection (LMD), ultra-high performance liquid chromatography-quadrupole/time-of-flight-mass spectrometry (UHPLC-Q/TOF-MS), and UHPLC coupled with triple quadrupole mass spectrometer (UHPLC-QqQ-MS) were applied to profile and determine flavonoids in various tissues in this study. Consequently, 43 peaks were detected by UHPLC-Q/TOF-MS, and 26 flavonoid compounds combined with seven triterpene compounds were identified or tentatively identified in the tissue extractions. The results indicated that the hydrophobic compounds, especially flavonoid or isoflavonoid aglycones and xanthone mainly accumulated in the cork, whereas the hydrophilic compounds, namely the flavonoid and isoflavonoid glycosides were usually found in the cortex or center (the part inside of endodermis). Samples of rhizomes from different growth ages and origins were simultaneously analyzed. It was shown that the bulb or lateral part of the rhizome generally possessed more total flavonoids than the vertical part or the primordium. The present study established a new practical method to evaluate the quality of the rhizome of B. chinensis and to explore the relationship between distribution patterns of secondary metabolites and growth years of plants, thus important information for cultivation and processing was provided.

Genetic diversity of endophytic fungi strains Fusarium proliferatum isolated from Belamcanda chinensis using ISSR and SRAP markers / 中草药

Objective: In order to understand the genetic diversity of endophytic fungi strain Fusarium proliferatum isolated from Belamcanda chinensis. Methods: A total of 52 ISSR primers and 90 SRAP primers were used to detect the genetic diversity among 17 F. proliferatum strains. Results: The results indicated that 27 ISSR primers and 38 SRAP primers were screened out for the genetic diversity analysis. 178 bands were amplified from 27 ISSR primers, among which 131 (63%) allelic variations were detected. However, 357 bands were amplified by 38 SRAP primers, among which 323 (91%) allelic variations were detected. The value of allelic polymorphism information content (PIC) of ISSR primers ranged from 0.19 to 0.91, with the average of 0.70 per primer. The value of PIC of SRAP primers ranged from 0.00 to 0.93, with the average of 0.72 per primer. The value of Nei's genetic similarity (GS) indexes of 17 strains based on ISSR, SRAP and ISSR + SRAP genetic locus varied from 0.73-0.99, 0.72-0.95 and 0.73-0.95, and with the average of 0.84, 0.85 and 0.85, separately. Cluster analysis showed that the 17 strains in this study could be clustered into three groups, three strains from the roots were clustered together, and F. proliferatum strains isolated from stems and leaves were gathered in other two groups. Cluster analysis revealed that genetic similarity of 17 strains were high, this suggested that the 17 strains had a near relationship, in accordance with the traditional morphology identification. Conclusion: The results show that the ISSR and SRAP technology is more efficient than traditional morphology identification. It is also found that ISSR and SRAP markers could more really reflect the genetic diversity of endophytic fungi strain F. proliferatum from B. chinensis, which can provide the basis for the application of molecular biotechnology in endophytic fungi of F. proliferatum from B. chinensis.

Rhizomatous induction of Belamcanda chinensis in vitro / 中草药

Objective To optimize the medium for rhizomatous induction of Belamcanda chinensis in vitro. Methods By plant tissue culture technology, the effects of various carbon source, NAA, and active carbon at different concentration on the rhizomatous formation of B. chinensis in vitro were studied. Results MS+6-BA 2.0 mg/L+NAA 0.5 mg/L+6% white sugar was the optimal medium for the rhiz-omatous formation of B. chinensis in vitro. Active carbon should not be added to the medium. The germination rate of rhizomatous in vitro was 61.03% on the MS + BA 2.0 mg/L+3% white sugar. Conclusion Sugar concentration is the main factor of the influence on the rhizomatous formation of B. chinensis in vitro.

Study on chemical constituents of Belamcanda chinensis (Ⅱ) / 中草药

ObjectTo investigate the chemical constituents in the rhizoma of Belamcanda chinensis (L.) DC. MethodsThe chemical constituents were isolated and purified by solvent extraction together with various chromatographic techniques. The structures were elucidated on the basis of chemical evidence and spectral data. ResultsThree compounds were isolated from the EtoAC extracts of the rhizome of B. chinensis which were isorhamnetin (Ⅹ), hispidulin (Ⅺ), dichotomitin ( ⅩⅡ); four compounds were isolated from n-BuOH extracts, which were iridin ( ⅩⅢ), tectoridin (ⅩⅣ), daucosterol ( ⅩⅤ), vittadinoside or stigmasterol-3-O-glucoside ( ⅩⅥ). ConclusionCompound Ⅺ is isolated from this medicinal plant for the first time.

Quantitative Determination of Mangiferin in Rhizoma Belamcandae and Its Substitute of Iris L / 中草药

Mangiferin, one of the active constituents of Rhizoma Belamcandae, in samples of Be-lamcanda chinensis (L.) DC. or its substitute was determined quantitatively by RP-HPLC. The 11 sam-ples collected from different localities for analysis were: 7 rhizomes of wildly grown or cultivated B. chi-nensis, 1 of its leaf and stem, and 3 substitutes (a wildly grown and another commercially available Iristectorum Maxim. and a I. dichotoma Pall. ). Results of the analysis showed that the contents of mangiferinin Rhizoma Belamcandae were significantly higher than that of its substitutes I. tectorum and I. di-chotoma. There were also certain significant differences between samples from different localities (P<0.05), but with no statistically significant difference between the rhizome or leaf and stem, neither be-tween cultivated and wildly grown samples, (P>0.05). The method was proved to be quick, simple andreproducible, and may provide a reliable basis for the quality control and evaluation of B. chinensis.