To obtain the immunomodulating polysaccharide from chaga mushroom (Inonotus obliquus sclerotia, IO), crude polysaccharide fractions (IO-M-CP and IO-CP, respectively) prepared from hot-water extract (IO-W) of I. obliquus by EtOH precipitation after MeOH reflux or not. After IO-W was re-dissolved in water followed by EtOH addition in the case without MeOH reflux, EtOH mixture was fractionated into EtOH-soluble (IO-E) and crude polysaccharide (IO-CP). In the meanwhile, MeOH-soluble fraction (IO-M) was separated from IO-W after MeOH reflux. The residue was dissolved in water and was added by EtOH, and then EtOH mixture was also fractionation into EtOH-soluble (IO-M-E) and crude polysaccharide (IO-M-CP). As a result of the macrophage stimulating activity of these fractions, IO-CP and IO-M-CP showed significantly increased cell proliferation and cytokines production than IO-W. Particularly, IO-M-CP promotes the production of IL-12 more than IO-CP. In the splenocytes proliferating activity and intestinal immune system modulating activity through Peyer’s patch, both of 2 crude polysaccharide fractions were significantly promoted in cell proliferation and cytokines production than IO-W, and IO-M-CP was more potent than IO-CP in IL-2 production from splenocytes and GM-CSF production (10 μg/mL) in Peyer’s patch cells. In addition, immunomodulating polysaccharide fractions (IO-M-CP and IO-CP) prepared from IO-W by EtOH precipitation with or without EtOH reflux showed no significant difference in the chemical composition and component sugar. These results suggested that MeOH reflux might exclude low-molecular weight materials from IO-W and consequently increase the immunomodulating activity of IO-M-CP. Therefore, it was confirmed that immunomodulation of polysaccharide prepared from hot-water extract of chaga mushroom was enhanced by fractionation including MeOH reflux and EtOH precipitation.

This study investigated the fermentative characteristics and immunomodulating activity in Kimchi added with various salts(salt replacement and herb-salt with Acanthopanax senticosus and Glycyrrhizae uralensis) for the reduction of Na concentration in Kimchi. Kimchi using a salt replacement and herb-salt showed a higher level of acidity (0.8～0.84%) than that of the control (0.7%) at 7-day fermentation. Kimchi using a salt replacement and herb-salt showed a lower level of salinity (1.72～1.98%) than that of control (2.3～2.57%) during fermentation. The growth of Lactobacillus spp. and Leuconostoc spp. recorded the highest level (2.3×10⁸ and 2.8×10⁶ cfu/g, respectively) in control at 6 day fermentation. However, those levels in Kimchi prepared with salt replacement and herb-salt were 3.5～5.4×10⁸ and 6.1×10⁶ cfu/g, respectively. It is assumed that the high level of acidity of Kimchi prepared with salt replacement and herb-salt was caused by the increase in the growth of Lactobacillus spp. and Leuconostoc spp.. When the macrophage stimulating activity of salt replacement kimchi (Salt-R kimchi) supplemented with hot-water extract from Acanthopanax sentisus (AS) or Glycyrrhiza uralensis (GU) was investigated on aging period, Salt-RA kimchi with AS 5% at 6 days (2.78-fold of saline control at 100 ㎍/㎖) and Salt-RG kimchi with GU 5% at 9 days (2.02-fold) significantly increased compared to the Salt-RA kimchi without AS or GU. In addition, Salt-RAG kimchi with AS 3% and GU 3% improved the bitter taste of Salt-RA and potently stimulated the macrophage at 6 days (1.28-fold of Salt-R kimchi) even though its activity was lower than Salt-RA (5%, 1.39-fold).

본 연구는 mushroom complete medium(MCM) 액체배지에 수삼 추출물(GE, 65°Bx)을 첨가하여 면역활성이 증진된 노루궁뎅이버섯(Hericium erinaceum) 균사체를 배양하고, 균사체로부터 활성다당성분을 분획하고자 하였다. MCM에 대하여 GE를 5, 10과 15%(v/v) 첨가한 액체배지에서 균사체를 배양하고, 각각의 조다당획분(HE-GE-5-CP, HE-GE-10-CP와 HE-GE-15-CP)으로 분획하여 면역활성을 측정한 결과, HE-GE-10-CP는 HE-GE-5-CP와 HE-GE-15-CP보다 높은 활성을 나타내었으며, GE를 첨가하지 않은 MCM에서 배양된 균사체 조다당획분(HE-CP)보다 유의적으로 증진된 면역활성을 나타내었다. 또한, HE-GE-10-CP의 DEAE-Sepharose CL-6B 분획물 중 가장 높은 활성을 나타낸 HE-GE-10-CP-II획분은 대조군인 HE-CP의 어떠한 획분보다도 유의적으로 높은 면역활성과 암 전이 억제활성을 나타내었다. 한편, 활성획분인 HE-GE-10-CP-II는 arabinose, rhamnose, galactose, glucose와 uronic acid(molar ratio; 0.34:0.26:0.99:1.00:0.39)로 구성되어 있으나, 대조군인 HE-CP의 동일용매 용출획분으로서 HE-GE-10-CP-II보다는 활성이 낮은 HE-CP-II는 fucose, mannose, galactose와 glucose(molar ratio; 0.32:0.55:1.00:0.96)를 함유하여 다른 구성당 분포를 나타내었다. 따라서 노루궁뎅이버섯 균사체 액체배양에서 수삼 추출물 첨가는 균사체의 구성당 변화를 통한 면역활성 증진에 관여하는 것으로 사료되어 기능성 소재 개발에 유용할 것으로 사료된다.

This study was designed to evaluate a repeated oral dose toxicity and immunomodulating activity of Pulsatilla koreana and Artemisiae annuae in Sprague-Dawley rats. The female rats were treated with Pulsatilla koreana and Artemisiae annuae of control group, low group (0.5 ml/kg), medium group (1 ml/kg), high group (2 ml/ kg) for distilled water, intragastrically for 4 weeks, respectively. To ensure the safety of Pulsatilla koreana and Artemisiae annuae such as the following were observed and tested. We examined the body weight, the feed intake, the clinical signs, the ophthalmological test, the hematological and the serum biochemical analysis. We also observed the histopathological changes of liver and kidney in rats. Hematological results were the increase of neutrophils, lymphocytes and monocytes in the high dose group of Pulsatilla koreana. The increase immune cells in the high dose group of Pulsatilla koreana might immunomodulating activity. No significant differences in body weight, feed intake, serum biochemical analysis and histopathological between control and fed group were found. In conclusion, Pulsatilla koreana and Artemisiae annuae is physiologically safe and improve immunomodulating activity.