• Korean Journal of Medicinal Crop Science
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• v.9 no.3
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• pp.205-210
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• 2001

Dried parts of the two species are difficult to distinguish morphologically, thus Codonopsis radix has been sold instead of Adenophorae radix in herbal medicine market. Therefore, this study was conducted to develop the genetic marker through the examination of the phylogenetic relationships between two Adenophora triphylla(Thunb.) A. DC. var. japonica Hara, two Adenophora radiatifolia Nakai, five Codonopsis lanceolata(Sieb. et Zucc)Trautv. using RAPD analysis. Fifty decarmer oligonucleotide primers were screened for the RAPD analysis, and four primers generated distinct RAPD markers specific to Adenophorae radix and Codonopsis radix. Based on the RAPD patterns, the genetic relationships between three herbal medicine were analyzed by UPGMA method. As a result, Adenophorae radix and Codonopsis radix were classified into two major subgroups on the basis of the genetic similarity coefficient. The specific RAPD patterns generated by the selected primers were reproducible from dried materials. Furthermore, the specific RAPD patterns were produced from the mixture of dried roots of A. triphylla and C. lanceolata. These results prone the usefulness of the RAPD analysis for the discrimination of pure materials from the mixtures of A. triphylla and C. lanceolata.

• Journal of Physiology & Pathology in Korean Medicine
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• v.34 no.3
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• pp.136-141
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• 2020

The purpose of this study was to investigate the action mechanism of the roots of Adenophora triphylla var. japonica extract (ATE) in 3T3-L1 adipocytes. Cell toxicity test by MTT assay and lipid accumulation was performed to evaluate the inhibitory effect on the differentiation of adipocyte from preadipocytes induced by MDI differentiation medium, while adipogenesis related proteins expression level were evaluated by western blotting. As a result, ATE inhibited MDI-induced adipocyte differentiation in 3T3-L1 cells dose-dependently without cytotoxicity. Our results showed that ATE inhibited the phosphorylation of IRS1, thereby decreasing the expression of PI3K110α and reducing the phosphorylation of AKT and mTOR, resulting in attenuated protein expression of C/EBPα, PPARγ, ap2 and FAS in 3T3-L1 cells. These results suggest anti-adipogenic functions for ATE, and identified IRS1 as a novel target for ATE in adipogenesis.

• Korean Journal of Medicinal Crop Science
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• v.3 no.1
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• pp.66-70
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• 1995

These experiments were conducted to investigate the growth and characteristics of Adenophora triphylla DC. seed, and effects of some pretreatments and light condition on the gemination of it. The results were as follows. Seed coat was soft and white at 30 days after flowering, but it became hard and brown from 40 days. The 1,000 grain weight was 247mg at 40 days after flowering and high at 50 days as 268mg, and decreased from 60 days. Germination percentage was highest at 100ppm $GA_3$ treatment for 24 hours as 96.7% and 88.3% at moistured treatment at $0^{\circ}C$ for 7 days, but only 40% at control. Adenophora triphylla. seed was light germinator, but it could be germinated 80% even under dark condition if $GA_3$ was treated 100ppm for 24 hours. The optimum temperature of germination in Adenophora triphylla. seeds was $25^{\circ}C$.

• Korean Journal of Medicinal Crop Science
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• v.25 no.6
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• pp.411-417
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• 2017

Background: Adenophora triphylla var. japonica (Regel) H. Hara shows vegetative growth with radical leaves during the first year and shows reproductive growth with cauline leaves and bolting during the second year. In addition, the shape of the plant varies within the same species. For this reason, there are limitations to classifying the species by visual examination. However, there is not sufficient genetic information or molecular tools to analyze the genetic diversity of the plant. Methods and Results: Approximately 34.59 Gbp of raw data containing 342,487,502 reads was obtained from next generation sequencing (NGS) and these reads were assembled into 357,211 scaffolds. A total of 84,106 simple sequence repeat (SSR) regions were identified and 14,133 primer sets were designed. From the designed primer sets, 95 were randomly selected and were applied to the genomic DNA which was extracted from five plants and pooled. Thirty-nine primer sets showing more than two bands were finally selected as SSR markers, and were used for the genetic relationship analysis. Conclusions: The 39 novel SSR markers developed in this study could be used for the genetic diversity analysis, variety identification, new variety development and molecular breeding of A. triphylla.

• Korean Journal of Plant Resources
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• v.9 no.2
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• pp.171-175
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• 1996

The objective of this study is to promote germination for mass cultivation of Adenophora triphylla. After the seed soaking for I day in BA and $GA_3$ solution, seed germination was effectively promoted in the treatment of 500mg/L $GA_3$ and the highest germination rate was 94%.The average germination days were 1.5days earlier than those of non-treated seeds. In a chemicals treatment, seed germination rate was 54% in 1% $KNO_3$ treatment but KOH treatment was no significantly effected. The durations of chilling treatment for breaking of dormancy were longer than 2weeks. The optimal temperature for germination was $25^{\circ}C$. The promotion of seed germination is presumed to be due to the breaking of dormancy by $GA_3$, chilling treatment rather than seed coat maceration by KOH or $KNO_3$ treatments.

• Journal of Pharmacopuncture
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• v.22 no.4
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• pp.253-259
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• 2019

Objectives: It is reported that tumor-associated macrophages (TAMs) contribute to cancer progression by promoting tumor growth and metastasis. The purpose of this study is to investigate the effect of different fractions of Adenophora triphylla var. japonica (AT) on the polarization of macrophages into the M2 phenotype, a major phenotype of TAMs. Methods: We isolated hexane, ethyl acetate, and butanol fractions from crude ethanol extract of AT. The cytotoxicity of AT in RAW264.7 cells was examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. RAW264.7 cells were polarized into the M2 phenotype by treatment with interleukin (IL)-4 and IL-13. The expression of M2 macrophage marker genes was detected by reverse transcription polymerase chain reaction (RT-PCR). The phosphorylation level of signal transducer and activator of transcription 6 (STAT6) was investigated by western blot analysis. The migration of Lewis lung carcinoma (LLC) cells was examined by transwell migration assay using conditioned media (CM) collected from RAW264.7 cells as a chemoattractant. Results: Among various fractions of AT, the ethyl acetate fraction of AT (EAT) showed the most significant suppressive effect on the mRNA expression of M2 macrophage markers, including arginase-1, interleukin (IL)-10 and mannose receptor C type 1 (MRC-1), up-regulated by treatment of IL-4 and IL-13. In addition, EAT suppressed the phosphorylation of STAT6, a critical regulator of IL-4 and IL-13-induced M2 macrophage polarization. Finally, the increased migration of Lewis lung carcinoma (LLC) cells by CM from M2-polarized RAW264.7 cells was reduced by CM from RAW264.7 cells co-treated with EAT and M2 polarization inducers. Conclusion: We demonstrated that EAT attenuated cancer cell migration through suppression of macrophage polarization toward the M2 phenotype. Additional preclinical or clinical researches are needed to evaluate its regulatory effects on macrophage polarization and anti-cancer activities.

• Journal of Physiology & Pathology in Korean Medicine
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• v.32 no.3
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• pp.157-164
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• 2018

This study was designed to investigate the mechanism of the synergistic anticancer effect of Astragalus membranaceus (AM) and Adenophora triphylla var. japonica (AT) in H1299 human lung carcinoma cells. A combined treatment of ethanol extract of AM (EAM) and AT (EAT) explosively increased the reactive oxygen species (ROS) generation in H1299 cells compared to the single treatment of each of them. Co-treatment of N-acetyl-L-cysteine (NAC) with EAM and EAT markedly enhanced the cell viability and suppressed apoptosis in H1299 cells, suggesting that ROS generation contributed to the anticancer effect of EAM and EAT. Interestingly, the combined treatment of EAM and EAT down-regulated p-AKT in H1299 cells, which was abrogated by NAC treatment. These results clearly indicated that ROS generation mediated the inactivation of AKT. Co-treatment of LY294002 with EAM and EAT significantly reduced the cell viability at a concentration which EAM and EAT didn't show any cytotoxicity. In addition, the recovery of cell viability by co-treatment of NAC with EAM and EAT was quite reversed by LY294002 treatment, which confirmed that the inactivation of AKT played a pivotal role in ROS-mediated apoptosis. Taken together, our results demonstrated that the synergistic anticancer effect of EAM and EAT was mediated by ROS generation and inactivation of AKT. We provide a valuable preclinical data for the development of more effective combination of AM and AT to treat lung cancer.

• Korean Journal of Plant Resources
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• v.30 no.4
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• pp.352-363
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• 2017

To investigate skin-whitening effect of Adenophora triphylla var. japonica sprout extract, antioxidant activity, inhibitory effect on tyrosinase and melanin synthesis in B16/F10 melanoma cell were examined. Total phenolic content (246.25 mg GAE/g) and total flavonoid content (303.94 mg RE/g) of ethyl acetate fraction from Adenophora triphylla sprout (EFAT) showed the highest contents than other fractions (n-hexane, chloroform and distilled water). Antioxidant activities of EFAT has been evaluated using ABTS, DPPH radical scavenging activities, FRAP and inhibitory effect of lipid peroxidation. EFAT showed excellent radical scavenging activity and inhibitory effect on MDA production. Inhibitory effect of tyrosinase as a major enzyme of melanin synthesis was also measured. In these results, EFAT showed higher inhibitory effect against L-DOPA (51.27%) than L-tyrosine. $IC_{50}$ value on ${\alpha}-glucosidase$ was $41.93{\mu}g/ml$. In B16/F10 melanoma cells, EFAT inhibited melanin synthesis at $200{\mu}g/ml$ concentration (about 42% decrease). Finally, main physiological compounds of EFAT were identified as a rutin and a chlorogenic acid using high performance liquid chromatography.

• Journal of Bio-Environment Control
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• v.28 no.4
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• pp.404-410
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• 2019

This study was performed to investigate the seed morphological characteristics and dormancy type of Adenophora triphylla var. japonica Hara that high valued medicinal crop and to select the disinfectants and light quality for germination rate improvement. The seed disinfection was carried out using distilled water (control), NaClO 4%, $H_2O_2$ 4%, and benomyl $500mg{\cdot}L^{-1}$. The light quality treatments were set to dark condition (control I), fluorescent lamp (control II), LEDs [red, blue, green, and combined RB LEDs (red:blue = 8:2, 6:4, 4:6, 2:8)] with a photoperiod of 12/12 (light/dark) and light intensity $150{\pm}10{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ photosynthetic photon flux density. Although the Adenophora triphylla var. japonica Hara seed was an underdeveloped embryo (E) and seed (S) with an embryo (E):seed (S) ratio of 0.4, it is germinated within 30 days, and seed moisture saturation was reached within 6 hours after immersion. After seed disinfection, the mold incidence rate was significantly inhibited, and the final germination rate was the highest at 87% in the benomyl seed disinfection. The final germination rate was the highest at 92% in the red light, and the mean daily germination was the lowest in the R2B8. Therefore, there is almost no dormancy in the Adenophora triphylla var. japonica Hara seed, and benomyl seed disinfectant and red light were effective in the improvement of germination rate. So it is considered to the high value of use for medicinal crop Adenophora triphylla var. japonica Hara cultivation.

• Natural Product Sciences
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• v.20 no.4
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• pp.243-250
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• 2014

A simple GC method with a FID detector was developed in order to determine two main compounds (${\beta}$-sitosterol and lupenone) for Adenophorae Radix. ${\beta}$-Sitosterol and lupenone were analyzed by the gradient thermal ramping method. Nitrogen was used as the carrier gas at 108 kPa. The flow rate of gas was 2.0 mL/min; $2{\mu}L$ of filtered sample was injected at a split ratio of 1 : 80. This method was fully validated with respect to linearity, precision, accuracy and robustness. Further, this GC-FID method was applied successfully in order to quantify two compounds in an Adenophorae Radix extract. The GC analytical method for classification analysis was performed by repeated analysis of 59 reference samples in order to differentiate between Adenophora triphylla var. japonica Hara and 14 Codonopsis lanceolata. The results indicate that the GC-FID method is suitable and reliable for the quality evaluation of Adenophorae Radix.