Evidence-guided optimization of herbal formula Cheong-Hwa-Bo-Um-Tang using multiplex cytokine profiling

Background: Cheong-Hwa-Bo-Um-Tang (CHB) has been traditionally used to treat pharyngolaryngitis and throat inflammation in Korea. However, the anti-inflammatory activity of CHB and the exact component proportion ratio of CHB have not been fully reported.Materials and Methods: This study was conducted to optimize the component proportion ratio of CHB on the basis of experimental evidence using multiplex cytokine profiling assay in LPS-stimulated RAW 264.7 mouse macrophages.Results: The optimized CHB (named Chunhwabun-Cheong-Hwa-Bo-Um-Tang) inhibited the production of NO, IL-6, MCP-1, GMCSF, IL-10, IP-10, VEGF, M-CSF, RANTES, MIP-1β, LIF, LIX, and TNF-α in LPS-stimulated RAW 264.7 mouse macrophages without decreasing cell viability (P<0.05). Chunhwabun-Cheong-Hwa-Bo-Um-Tang consisted of roots of Scrophularia buergeriana, Trichosanthes kirilowii, Anemarrhena asphodeloides, Phellodendron amurense, Angelica gigas, Rehmannia glutinosa, Paeonia japonica, Cnidium officinale, and Glycyrrhiza uralensis (in the ratio of 8: 3: 3: 3: 3: 4: 4: 3: 3).Conclusion: The results of this study indicate that the multiplex cytokine profiling assay could be one of the effective methods for evidence-guided optimization of the component proportion ratio of a traditional herbal formula used for treating excessive inflammation such as the cytokine storm.Key words: Cheong-Hwa-Bo-Um-Tang; Lipopolysaccharide; Inflammation; Macrophages; Cytokine; Nitric Oxid

Scientific Evidence for Korean Medicine and Its Integrative Medical Research 2017

In 2016, the total medical fee for Korea Medicine (KM) in South Korea was over 4 billion dollars and the National Health Insurance of South Korea paid about 2.09 billion dollars for medical services provided by 19, 737 KM doctors in 302 KM hospitals and 14, 150 KM local clinics. Actually, 12.9 million Korean patients received 105 million outpatient services provided by KM doctors in 2016. The older the Korean society is, the more important the role of KM becomes, because Korea is anticipated to be a hyperaged society in 2028. As well as the Donguibogam, which was enlisted on the Memory of the Word by UNESCO in 2009, Sasang typology, Sasang Constitutional Medicine, Sa-am acupuncture, Chuna therapy, Pharmacopuncture, Korean physical therapy, Korean psychotherapy, and so on characterize KM. But more integrative researches and scientific evidences for KM might strengthen both clinical efficacy and applicability of KM. In this respect, it seems to be unavoidable that KM utilizes cutting-edge techniques of modern science. The more accurate and efficient practice of KM innovated with the modern bioscientific technology would help people live healthy lives. Our special issue, which had opened for 6 months in the first half of 2017, focused on scientific evidence for KM and its integrative medical research

Intercultural Usage of Mori Folium: Comparison Review from a Korean Medical Perspective

Objectives. A review on studies related to the use of Mori folium, the leaves of Morus alba, was conducted with the goal of identifying new clinical applications in Korean medicine. Methods. Global literature search was conducted using three electronic databases up to January 2015 with the term Morus alba and its Korean terms. KM literatures including textbooks and standard pharmacopoeia were separately hand-searched and reviewed to provide comparison. Data were extracted according to predetermined criteria, and clinical uses were standardized with ICD-10 categories. Results. 159 potentially relevant studies were identified, and 18 articles including 12 ethnopharmacologic and 6 clinical studies were finally included in this analysis. Ethnopharmacologic studies from 8 countries provided 17 clinical uses. We found that five out of six clinical trials were related to diabetes and suggested a moderate short-term to mild long-term effect. And 43 Korean texts also provided 156 clinical uses in 35 categories including ocular and respiratory disorders. Discussion and Conclusions. Though majority of the clinical uses were also found in Korean medicine literature, treatment of infertility, jaundice, cognitive disorder, and hyperpigmentation was found to be effective and diabetes with Morus alba was recognized to have clinical importance

Ethanol extract of Scutellaria baicalensis Georgi prevents oxidative damage and neuroinflammation and memorial impairments in artificial senescense mice

Aging is a progressive process related to the accumulation of oxidative damage and neuroinflammation. We tried to find the anti-amnesic effect of the Scutellaria baicalens Georgia (SBG) ethanol extract and its major ingredients. The antioxidative effect of SBG on the mice model with memory impairment induced by chronic injection of D-galactose and sodium nitrate was studied. The Y-maze test was used to evaluate the learning and memory function of mice. The activities of superoxide dismutase, catalase and the content of malondialdehyde in brain tissue were used for the antioxidation activities. Neuropathological alteration and expression of bcl-2 protein were investigated in the hippocampus by immunohistochemical staining. ROS, neuroinflammation and apoptosis related molecules expression such as Cox-2, iNOS, procaspase-3, cleaved caspase-3, 8 and 9, bcl-2 and bax protein and the products of iNOS and Cox-2, NO, PGE2, were studied using LPS-activated Raw 264.7 cells and microglia BV2 cells. The cognition of mice was significantly improved by the treatment of baicalein and 50 and 100 mg/kg of SBG in Y-maze test. Both SBG groups showed strong antioxidation, antiinflammation effects with significantly decreased iNOS and Cox-2 expression, NO and PGE2 production, increased bcl-2 and decreased bax and cleaved caspase-3 protein expression in LPS induced Raw 264.7 and BV2 cells. We also found that apoptotic pathway was caused by the intrinsic mitochondrial pathway with the decreased cleaved caspase-9 and unchanged cleaved caspase-8 expression. These findings suggest that SBG, especially high dose, 100 mg/kg, improved the memory impairments significantly and showed antioxidation, antiinflammation and intrinsic caspase-mediated apoptosis effects

INHIBITORY EFFECT OF EMODIN ON RAW 264.7 ACTIVATED WITH DOUBLE STRANDED RNA ANALOGUE POLY I:C

Background: Emodin (3-methyl-1, 6, 8-trihydroxyanthraquinone) is a compound which can be found in Polygoni Multiflori Radix (PMR). PMR is the root of Polygonum multiflorum. PMR is used to treat dizziness, spermatorrhea, sores, and scrofula as well as chronic malaria traditionally in China and Korea. The anti-tumor property of emodin was already reported. However, anti-viral activity of emodin on macrophages are not fully reported. Materials and Methods: Effects of emodin on RAW 264.7 mouse macrophages induced by polyinosinic-polycytidylic acid (poly I:C), a synthetic analog of double-stranded RNA, were evaluated. Results: Emodin restored the cell viability in poly I:C-induced RAW 264.7 at concentrations of up to 50 μM. Emodin significantly inhibited the production of nitric oxide, IL-1α, IL-1β, IL-6, GM-CSF, G-CSF, M-CSF, MCP-1, MIP-1α, MIP-1β, MIP-2, RANTES, and IP-10 as well as calcium release and mRNA expression of signal transducer and activated transcription 1 (STAT1) in poly I:C-induced RAW 264.7 (P < 0.05). Conclusion: This study shows the inhibitory effect of emodin on poly I:C-induced RAW 264.7 via calcium-STAT pathway

Effect of Lactobacillus pentosus-Fermented Artemisiae Argi Folium on Nitric Oxide Production of Macrophage impaired with Various Toxicants

Objectives : The purpose of this study is to investigate the effect of Water Extract from Lactobacillus pentosus-fermented ARTEMISIAE ARGI FOLIUM (AFL) on nitric oxide production of mouse macrophage Raw 264.7 cells impaired by various toxicants such as gallic acid, EtOH, nicotine, acetaminophen, and acetaldehyde. Methods : ARTEMISIAE ARGI FOLIUM was fermented with Lactobacillus pentosus and extracted by water. Nitric oxide production of mouse macrophage Raw 264.7 cells was measured by Griess reagent assay. Examined concentrations of AFL were 10, 50, 100, 200, 400 ug/mL. Results : The results of the experiment are as below. 1. AFL at the concentration of 400 ug/mL significantly recovered nitric oxide production which was reduced by gallic acid (100 uM) in Raw 264.7 cells. 2. AFL at the concentration of 200, 400 ug/mL significantly recovered nitric oxide production which was reduced by EtOH (100 uM) in Raw 264.7 cells. 3. AFL at the concentration of 400 ug/mL significantly recovered nitric oxide production which was reduced by nicotine (1mM) in Raw 264.7 cells. 4. AFL at the concentration of 200, 400 ug/mL significantly recovered nitric oxide production which was reduced by acetaminophen(2mM) in Raw 264.7 cells. 5. AFL at the concentration of 200, 400 ug/mL significantly recovered nitric oxide production which was reduced by acetaldehyde (200 uM) in Raw 264.7 cells. Conclusions : AFL could be supposed to have the immune-enhancing activity related with nitric oxide production of macrophage impaired by various toxicants

Immunomodulatory Effects of Lycii Radicis Cortex Water Extract on Lipopolysaccharide-Activated RAW 264.7 Mouse Macrophages

The root bark of Lycium chinense Miller (Solanaceae), also known as Lycii radicis cortex, is one of famous plant-originated drugs in Korean medicine owing to its anti-tussive, anti-asthmatic, and antipyretic properties. In this study, the effect of Lycii radicis cortex water extract (LRC; 10 - 200μg/mL) on inflammatory mediators from lipopolysaccharide (LPS)-activated RAW 264.7 mouse macrophages was inspected. After 24 h incubation with LRC, cell viability, nitric oxide (NO), and various cytokines from RAW 264.7 were measured. LRC increased the cell viability of RAW 264.7 at concentrations of up to 200 μg/mL. LRC significantly inhibited the production of NO, granulocyte colony-stimulating factor (G-CSF), tumor necrosis factor-α (TNF-α), platelet derived growth factor-BB (PDGF-BB), interleukin (IL)-2, and IL-10 in LPS-activated RAW 264.7 (P &lt; 0.05). As well, LRC diminished the production of granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide-induced CXC chemokine (LIX). The current results suggest that LRC has immunomodulatory property to alleviate excessive immune reactions during the activation of macrophages by LPS

Inhibitory Effect of Gallic acid on Production of Interleukins in Mouse Macrophage Stimulated by Lipopolysaccharide

Objectives: Gallic acid (GA) is the major component of tannin which could be easily founded in various natural materials such as green tea, red tea, grape juice, and Corni Fructus. The purpose of this study is to investigate the effect of Gallic acid (GA) on production of interleukin (IL) in mouse macrophage Raw 264.7 cells stimulated by lipopolysaccharide (LPS). Methods: Productions of interleukins were measured by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on xMAP (multi-analyte profiling beads) technology. Firstly, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, it was incubated with the antibody-conjugated beads for 30 minutes. And detection antibody was added and incubated for 30 minutes. And Strepavidin-conjugated Phycoerythrin (SAPE) was added. After incubation for 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System and concentration of interleukin was determined. Results: The results of the experiment are as follows. 1. GA significantly inhibited the production of IL-3, IL-10, IL-12p40, and IL-17 in LPS-induced mouse macrophage RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). 2. GA significantly inhibited the production of IL-6 in LPS-induced mouse macrophage RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). 3. GA diminished the production of some cytokine such as IL-4, IL-5, and IL-13 in LPS-induced mouse macrophage RAW 264.7 cells. 4. GA did not show the inhibitory effect on the production of IL-1αand IL-9 in LPS-induced mouse macrophage RAW 264.7 cells. Conclusions: These results suggest that GA has anti-inflammatory activity related with its inhibitory effects on the production of interleukins such as IL-3, IL-10, IL-12p40, IL-17, and IL-6 in LPS-induced macrophages