Jung-Hoon Kim

Traditional herbal formula Jakyakgamcho-tang (Paeonia lactiflora and Glycyrrhiza uralensis) impairs inflammatory chemokine production by inhibiting activation of STAT1 and NF-κB in HaCaT cells

A traditional herbal formula Jakyakgamcho-tang (JYGCT; Paeonia lactiflora and Glycyrrhiza uralensis) has been used for treatment of backache, muscle pain, acute abdominal pain, neuralgia, bronchial asthma, and painful peripheral neuropathy in Oriental medicine. We report on our experiments using the HaCaT human keratinocyte cell line showing that a traditional herbal formula JYGCT has inhibitory effects on inflammatory responses in skin. Stimulation with tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) caused a significant increase in the production of the following chemokines: thymus- and activation-regulated chemokine (TARC)/CCL17; macrophage-derived chemokine (MDC)/CCL22; regulated on activation, normal T-cell expressed and secreted (RANTES)/CCL5; and interleukin-8 (IL-8) in HaCaT cells. By contrast, treatment with JYGCT extract significantly reduced the production of TARC, MDC, RANTES, and IL-8, but caused no cytotoxicity, compared with TNF-α and IFN-γ-treated control cells. Consistently, JYGCT extract downregulated the mRNA expression of TARC, MDC, RANTES, and IL-8 induced by TNF-α and IFN-γ in a dose-dependent manner. In addition, TNF-α and IFN-γ markedly increased the phosphorylation of signal transducer and activator of transcription 1 (STAT1) and the nuclear translocation of nuclear factor kappa B (NF-κB) in HaCaT cells. By contrast, TNF-α and IFN-γ-induced activation of STAT1 and NF-κB activation was inhibited by JYGCT treatment in a dose-dependent manner. Our data indicate that JYGCT attenuates TNF-α and IFN-γ-mediated chemokine production by targeting the STAT1 and NF-κB signalling in keratinocytes. Our findings suggest that JYGCT has potential as a therapeutic drug candidate for the treatment of inflammatory skin diseases.

Anti-Inflammatory Effects of Artemisia Leaf Extract in Mice with Contact Dermatitis In Vitro and In Vivo

The leaves of Artemisia argyi Lev. et Vant. and A. princeps Pamp. are well known medicinal herbs used to treat patients in China, Japan, and Korea with skin problems such as eczema and itching, as well as abdominal pain and dysmenorrhoea. We investigated the anti-inflammatory effects of Artemisia leaf extract (ALE) using CD mice and Raw 264.7 cells. The effects of ALE on histopathological changes and cytokine production in ear tissues were assessed in mice with CD induced by 1-fluoro-2,4-dinitrobenzene (DNFB). Moreover, the anti-inflammatory effects on production levels of prostaglandin E2 (PGE2) and nitric oxide (NO) and expression levels of cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) were investigated in Raw 264.7 cells. Topical application of ALE effectively prevented ear swelling induced by repeated DNFB application. ALE prevented epidermal hyperplasia and infiltration of immune cells and lowered the production of interferon- (IFN-) gamma (γ), tumour necrosis factor- (TNF-) alpha (α), and interleukin- (IL-) 6 in inflamed tissues. In addition, ALE inhibited expression of COX-2 and iNOS and production of NO and PGE2 in Raw 264.7 cells. These results indicate that Artemisia leaf can be used as a therapeutic agent for inflammatory skin diseases and that its anti-inflammatory effects are closely related to the inhibition of inflammatory mediator release from macrophages and inflammatory cytokine production in inflamed tissues.

Simultaneous quantification and antiatherosclerosis effect of the traditional Korean medicine, Hwangryunhaedok-tang

BackgroundHwangryunhaedok-tang (HHT) is a traditional herbal medicine that is used for the treatment of fever, inflammation, gastritis, and hypertension. In this study, we performed simultaneous determination of the five components, geniposide (1), baicalin (2), coptisine (3), palmatine (4), and berberine (5) in HHT by using a high-performance liquid chromatography–photodiode array (HPLC–PDA) analysis. We also evaluated the antioxidative activity of HHT and compounds 1–5 by measuring their effects on low-density lipoprotein (LDL) oxidation and antiproliferative abilities in vascular smooth muscle cells (VSMCs).MethodsFive compounds were separated within 40xa0min by using a Gemini C18 column (temp. 35°C; two-component gradient elution; flow rate 1.0xa0mL/min; detector 240 and 277xa0nm). The activities of HHT and compounds 1–5 were tested with the radical scavengers 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt and 2,2-diphenyl-1-picrylhydrazyl, in thiobarbituric acid reactive substance assays, and in relative electrophoretic mobility assays using CuSO4-induced LDL oxidation systems. The antiproliferative effects of samples on platelet-derived growth factor (PDGF)-induced VSMC proliferation were studied by using a cell proliferation assay.ResultsRegression analysis of the five major compounds showed good linearity (r2u2009≥u20090.9997) in different concentration ranges. The recoveries of the five compounds were in the range 86.31–110.78%, with relative standard deviations below 2.1%; those of intra- and interday precision were 0.04–3.78% and 0.04–1.69%, respectively. HHT reduced the oxidation properties of LDL induced by CuSO4 and inhibited cell proliferation in PDGF-treated VSMCs. Among the five components, compound 2 could effectively suppress LDL oxidation and PDGF-induced VSMC proliferation.ConclusionsThe established HPLC–PDA method will help to improve quality control of HHT. The results demonstrate that HHT has antiatherosclerotic activity and that it functions by modulating LDL oxidation and VSMC proliferation. The effects of HHT may be attributed, at least I part, to compound 2.

Kochia scoparia induces apoptosis of oral cancer cells in vitro and in heterotopic tumors

ETHNOPHARMACOLOGICAL RELEVANCEnKochia scoparia grows commonly in China, Japan, and Korea and its mature fruit has been used throughout the area in traditional medicine to treat diseases including skin problems and inflammatory and allergic disease. More importantly, Kochia scoparia has been prescribed to treat the malignant tumor of head and neck region and breast mass. Although it has been proposed as an anti-cancer agent for several cancers, its exact in vivo anti-cancer properties and the molecular mechanisms underlying its effects are poorly understood.nnnAIM OF THE STUDYnTo evaluate the anti-cancer activity of the methanol extract of K. scoparia, mature fruit (MEKS) on oral squamous cell carcinoma (OSCC) and to explore its mode of action.nnnMATERIALS AND METHODSnTo assess proliferation inhibition and apoptosis induction by MEKS, MTT assays, cell analysis, ANNEXIN V and PI double staining, and Hoechst 33342 staining were performed. The activation of caspases and the MAP kinase p38 was evaluated using Western blot analysis. The anti-cancer properties of MEKS in vivo were elucidated in a heterotopic OSCC animal model.nnnRESULTSnAfter OSCC cells were treated with MEKS, the numbers of sub-G1 accumulated cells and apoptotic bodies increased, indicating that MEKS inhibited OSCC cell proliferation selectively through induction of apoptosis. Apoptosis of MEKS-treated OSCC cells was induced in a dose-dependent manner by caspase-3 and -9 activation. In addition, pretreatment with p38 inhibitor SB203580 in combination with MEKS significantly prevented MEKS-induced apoptosis in OSCC cells and also decreased cleaved capase 3, 9, and cleaved PARP activity in western blotting. MEKS treatment significantly increased the apoptosis of OSCC and inhibited tumour growth in our animal model.nnnCONCLUSIONnTaken together, these results indicated that MEKS induced apoptosis of OSCC cells through caspase activation involving the p38 MAPK pathway. MEKS could be a promising anti-cancer candidate for OSCC treatment.

Influence of herbal combinations on the extraction efficiencies of chemical compounds from Cinnamomum cassia, Paeonia lactiflora, and Glycyrrhiza uralensis, the herbal components of Gyeji-tang, evaluated by HPLC method

During decoction process, the ingredients of herbal formula interact with each other, such that therapeutic properties and chemical extraction characteristics are altered. The crude drugs, Cinnamomum cassia (CC), Paeonia lactiflora (PL), and Glycyrrhiza uralensis (GU), are the main herbal constituents of Gyeji-tang, a traditional herbal formula. To evaluate the chemical interaction between CC, PL, and GU during the course of decoction, quantification of 16 marker compounds in the herbal decoction, performed using a Box-Behnken experimental design, was carried out by HPLC-diode array detection using validated method. Correlations between the amounts of marker compounds from CC, PL, and GU were assessed by multiple regression analysis. The results obtained showed that amounts of single herb marker compounds significantly changed (usually decreased) by decoction in the presence of other herbs and that these changes depended on the chemical natures of the markers and the herbal medicines present. Results also demonstrated that the extraction efficiencies of marker compounds increased when the proportion of the herb containing them was increased and decreased in proportion to amounts of herbs added. In conclusion, chemical interactions between compositional herbal medicines may occur when herbs are co-decocted. This study provides insight of understanding the herbal interactions in herbal formulae.

Effects of Lizhong Tang on gastrointestinal motility in mice

AIM To investigate the effects of Lizhong Tang, a traditional Chinese medicine formula, on gastrointestinal motility in mice. METHODS The in vivo effects of Lizhong Tang on GI motility were investigated by measuring the intestinal transit rates (ITRs) and gastric emptying (GE) values in normal mice and in mice with experimentally induced GI motility dysfunction (GMD). RESULTS In normal ICR mice, the ITR and GE values were significantly and dose-dependently increased by Lizhong Tang (ITR values: 54.4% ± 1.9% vs 65.2% ± 1.8%, P < 0.01 with 0.1 g/kg Lizhong Tang and 54.4% ± 1.9% vs 83.8% ± 1.9%, P < 0.01 with 1 g/kg Lizhong Tang; GE values: 60.7% ± 1.9% vs 66.8% ± 2.1%, P < 0.05 with 0.1 g/kg Lizhong Tang and 60.7% ± 1.9% vs 72.5% ± 1.7%, P < 0.01 with 1 g/kg Lizhong Tang). The ITRs of the GMD mice were significantly reduced compared with those of the normal mice, which were significantly and dose-dependently reversed by Lizhong Tang. Additionally, in loperamide- and cisplatin-induced models of GE delay, Lizhong Tang administration reversed the GE deficits. CONCLUSION These results suggest that Lizhong Tang may be a novel candidate for development as a prokinetic treatment for the GI tract.

Optimal processing conditions of Boswellia carteri Birdw. using response surface methodology

Background: Boswellia carteri Bridw. is being widely used for its anti-inflammatory properties, as well as for wound healing, antimicrobial, and immunomodulatory properties, and boswellic acids (BAs) are considered to be the main active constituents. Objectives: To investigate optimal conditions of stir-baking process for the resin of B. carteri with vinegar of using response surface methodology (RSM). Materials and Methods: The concentration of acetic acid, heating temperature, and heating time were set as influential factors, and the yields of chemical compounds were the response values which were optimally designed by a Box–Behnken design. The amounts of 11-keto-β-boswellic acid (KBA) and α-boswellic acid (αBA) in B. carteri resin were quantified using high-performance liquid chromatography analysis. Results: Maximum amounts of KBA and αBA in B. carteri resin were obtained using 6% acetic acid for 10 min at 90°C in preliminary test. Two factor interactions, such as acetic acid concentration–heating temperature and heating temperature–heating time, were significantly observed by multiple regression analysis. Optimal processing conditions from RSM were 5.83% for acetic acid concentration, 9.56 min for heating time, and 89.87°C for heating temperature. Under the modified conditions, the experimental value of the response was 11.25 mg/g, which was similar to the predicted value. Conclusions: The results suggest that the optimal conditions for the stir-baking process of B. carteri resin were determined by RSM, which was reliable and applicable to practical processing of herbal medicine. Abbreviations used: BAs: Boswellic acids; KBA: 11 keto β boswellic acid; αBA: α boswellic acid; BBD: Box–Behnken design; RSM: Response surface method; HPLC: High performance liquid chromatography; LOD: Limits of determination; LOQ: Limits of quantification; RSD: Relative standard deviation; ANOVA: Analysis of variance.

Extraction time and temperature affect the extraction efficiencies of coumarin and phenylpropanoids from Cinnamomum cassia bark using a microwave-assisted extraction method

Microwave-assisted extraction (MAE), an efficient extraction tool, was employed to extract a coumarin and five phenylpropanoids (cinnamic acid, cinnamyl alcohol, cinnamaldehyde, 2-hydroxycinnamadehyde, and 2-methoxycinnamaldehyde) from Cinnamomum cassia bark using water as the extraction solvent. Six marker compounds were quantified by high-performance liquid chromatography-diode array detector using a validated analytical method. To investigate the influences of temperature and time on the extraction yields of the six marker compounds, the water extracts of C. cassia bark were prepared using a MAE method at six different extraction temperatures (70, 75, 80, 85, 90, and 95°C) and times (2, 4, 6, 8, 10, and 12min). Their influences were assessed by multiple regression analysis. The results obtained demonstrated that higher extraction temperature and longer extraction time positively affected coumarin and cinnamyl alcohol contents, but negatively affected extract contents of cinnamic acid, cinnamaldehyde and 2-hydroxycinnamaldehyde (all p-<0.05). The extraction of 2-methoxycinnamaldehyde was affected by both positively and negatively by increasing temperature and time. These changes during MAE were assumed by the chemical natures of the marker compounds with various functional groups. In conclusion, temperature and times significantly affected the extraction efficiencies of a coumarin and five phenylpropanoids from C. cassia bark when a water-based MAE method was used. This study provides a novel approach to the preparation of the water extract of C. cassia bark using MAE.

Identification and monitoring of Korean medicines derived from Cinnamomum spp. by using ITS and DNA marker

In this study, we identified and evaluated the genetic relationships among Cinnamomum plants, which are used in traditional medicine. We also attempted to monitor the distribution of traditional medicines derived from Cinnamomum cassia by using DNA barcoding and a species-specific DNA marker. Plants of the genus Cinnamomum, and in particular C. cassia, are commonly used as medicinal herbs in the form of Cinnamomi Ramulus, Cinnamomi Cortex, and Cassiae Cortex Interior. However, it is difficult to distinguish among different Cinnamomum species based on morphological features, and so to overcome this limitation, nucleotide sequences of the internal transcribed spacer (ITS) region of Cinnamomum DNA were determined and compared. On the basis of the discrepancy in determined ITS sequences, a 408-bp product, amplified by the primer pair CC F1/CC R3, was developed as a C. cassia-specific DNA marker. Using the developed DNA marker in combination with the ITS 2 nucleotide sequence, we monitored imported and commercially supplied medicinal products derived from Cinnamomum plants in markets in Korean, China, and Japan. The results revealed that most of the specimens monitored were derived from C. cassia.

Optimization for decocting later of menthae herba in eungyo-san, a herbal formula, using response surface methodology with gas chromatography/mass spectrometry

Background: “Decocting later” is important procedure for the extraction of herbal medicines containing volatile compounds. Objective: This study was performed to investigate optimal conditions for “Decocting later” of Menthae herba in Eungyo-san (EGS) and correlation between extraction variables and the yields of d/l-menthol, a marker compound of Menthae herba. Materials and Methods: The decocting temperature, total decocting time, and decocting later time were chosen as individual variables, and the yield of d/l-menthol was set as the response value which were calculated by using a Box-Behnken design (BBD). The amount of d/l-menthol was quantified using gas chromatography/mass spectrometry. Results: Response surface methodology (RSM) was used to predict optimal conditions for decocting later of Menthae herba into the formula. Optimal conditions for “Decocting later” from RSM were as follows: 100.63°C of decocting temperature; 82.95 min of total decocting time; 19.11 min of decocting later time. Both decocting temperature and total decocting time showed significant correlation with the yield of d/l-menthol. Conclusions: These results suggest that the decocting temperature and total decocting time were influential factors, and RSM can be applied for optimizing the conditions of “Decocting later” of Menthae herba in EGS. Abbreviations used: KM: Korean medicine; EGS: Eungyo-san; GC/MS: Gas chromatography/mass spectrometry; RSM: Response surface methodology; SIM: Selected ion monitoring; LOD: Limits of determination; LOQ: Limits of quantification; RSD: Relative standard deviation; ANOVA: Analysis of variance; BBD: Box-Behnken design.