In a previous study, we fractionated crude polysaccharide (AME-CP) with macrophage-stimulating activity from a hot-water extract (AME) of Astragalus membranaceus. AME-CP contained glucose (Glc) as a main component sugar, suggesting that it might be rich in starch-like compounds (SLC). To enhance the immunostimulating activity of AME-CP by pruning SLC rarely known to contribute to activity, hydrolysate (AME-SH) was prepared by digesting with starch-related enzymes, including α-amylase and amyloglucosidase. AME-SH was found to contain substances with molecular weights ranging from 3.9 to 84.4 kDa. These substances were primarily composed of galactose, galacturonic acid, Glc, arabinose, rhamnose, and mannose. AME-SH significantly enhanced the production of macrophage-stimulating factors, including nitric oxide (NO), interleukin (IL)-6, and IL-12, in RAW 264.7 cells compared to AME-CP. Treatment of splenocytes isolated from C3H/HeN mice with AME-SH not only promoted IL-6 secretion, but also induced mitogenic activity. In addition, AME-SH promoted the secretion of hematopoietic growth factors including IL-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF) in Peyer's patch (PP) cells and stimulated bone marrow cell proliferation through these PP cells. In conclusion, hydrolysate (AME-SH) digested from AME-CP with starch-related enzymes could be used as a potential immunostimulant.

To increase industrial applicability of Astragalus membranaceus (AM) as immunostimulating materials, hot-water extract (AME) was prepared from AM and fermented with Kimchi-lactic acid bacteria (Lactobacillus sakei & Leuconostoc mesenteroides) to prepare fermented AM-postbiotics (FAME). Although FAME prepared from AM-postbiotics did not show a significant enhancement in macrophage stimulating activity compared to non-fermented AME, crude polysaccharide (FAME-CP) fractionated by EtOH precipitation from FAME showed significantly higher macrophage stimulating activity than AME-CP. Compared to AME-CP, FAME-CP showed dramatic changes in component sugar and molecular weight distribution. FAME-CP was a polysaccharide with a major molecular weight distribution of 113.4 kDa containing Man (44.2%), Glc (19.3%), Gal (10.2%), GalA (10.2%), and Ara (7.4%) as sugar components. FAME-CP with enhanced macrophage stimulatory activity not only increased expression levels of mRNA genes encoding macrophage-activated factors (iNOS, TNF-α, MCP-1, IL-6, and COX-2), but also led the nuclear translocation of activated p65 and c-Jun. In conclusion, crude polysaccharide from AM-postbiotics fermented with lactic acid bacteria could increase industrial applicability as a functional material with enhanced immunostimulating activity than AME-CP.

The major active components of Astragalus membranaceus (AM) are isoflavones, which exist in the form of various glycosides. Nuruk is a traditional fermentation starter in Korea, and is suitable for the biotransformation of isoflavone glycosides because it contains various microorganisms and enzymes. This study was performed to determine changes in the isoflavones and antioxidant properties of AM fermented (AF) with nuruk over 24 hours. AF was sampled after 0, 2, 4, 6, 12, 18, and 24 h of fermentation, and calycosin 7-glucoside, ononin, calycosin, and formononetin content, and the antioxidant properties of AF were analyzed. The total phenolic content increased with fermentation time, and the ABTS radical scavenging activity increased until 6 h of fermentation and then decreased. During fermentation, the isoflavone glycosides decreased significantly as fermentation time increased. The contents of calycosin and formononetin, which are aglycons of calycosin-7-glucoside and ononin, increased from 100.54 μg/g to 276.84 μg/g and from 56.29 μg/g to 123.04 μg/g, respectively, at 18 h of fermentation. Significant correlations were observed between fermentation time, isoflavone content, and antioxidant properties. The results of this study showed that fermentation with nuruk is suitable for the biotransformation of isoflavones in AM.

본 연구는 황기 에탄올 추출물과 황기 생물전환 추출물의 기능성 화장품 소재로서 가능성 여부를 규명하고자 하였다. 항산화 효과를 확인하기 위해 폴리페놀과 플라보노이드 함량을 측정하였고, 세포실험을 통해 피부 세포에 대한 독성과 피부 세포 노화에 대한 연구를 수행하였다. 항산화 실험 결과, 폴리페놀, 플라보노이드 효과에서는 폴리페놀과 플라보노이드의 함량이 농도 의존적으로 증가하였고, 생물전환 추출물에 비해 에탄올 추출물에서 높은 것으로 확인되었다. 황기 추출물과 생물전환된 황기추출물이 HDF 세포에서 농도 의존적으로 MMP1 발현 억제 효과가 증가되는 것을 확인하였다. 따라서 본 연구는 황기 추출물과 황기 생물전환 추출물의 기능성 화장품 소재로 사용 시 항산화와 피부 노화 예방 관점에서의 기능성 화장품 소재로 매우 유용하게 활용될 수 있을 것으로 사료된다.

본 연구는 배지의 종류에 따른 더덕과 황기의 발아와 육묘기의 생육을 구명하기 위해 수행되었다. 종자는 상업적 상토(토실이), 코이어, 암면, 우레탄스펀지에 파종하였다. 파종 후 밀폐형 식물생산시스템에서 22일간 발아 하였으며, 경상대학교 벤로형 유리온실에서 파종 후 35 일간 육묘 하였다. 양액은 EC 1.0dS·m-1, pH 6.5로 맞추 어 2일 간격으로 저면관수 하였다. 더덕의 최종발아율은 코이어와 암면 배지에서 각각 68.5%와 67.9%로 유의적으로 높았으며, 평균발아수 역시 코이어와 암면 배지에서 4.2립과 4.1립으로 가장 높았다. 황기의 경우 배지 종류에 따른 황기의 발아율은 유의적인 차이를 나타내지 않았다. 더덕의 초장과 엽면적은 암면 배지에서 각각 11.5cm와 11.3cm2로 유의적으로 높았으며, 최대근장과 뿌리의 생체중은 코이어 배지에서 5.8cm와 0.07g으로 가장 우수했다. 황기의 초장과 엽면적은 코이어 배지에서 14.0cm와 16.9cm2로 유의적으로 높았으며, 지하부의 생체중과 건물중은 0.34g과 0.03g으로 우레탄스펀지 배지에서 가장 높았다. 따라서 본 연구 결과에서 더덕과 황기의 발아와 육묘기의 생육을 위해 암면과 코이어 배지가 적합한 것으로 판단된다.

The feed value of Astragalus membranaceus leaves and straws was amended by fermentation using effective microorganisms, such as molasses (T1), Rhodobacter capsulatus (T2), Bacillus subtilis (T3), Lactobacillus acidophilus (T4), or Saccharomyces cerevisiae (T5), with no supplements in the control (C). The crude protein (CP), ether extract (EE), crude fiber (CF), neutral detergent fiber (NDF), and acid detergent fiber (ADF) contents in the fermentation-treated A. membranaceus leaves decreased, whereas nitrogen free extract (NFE) content increased significantly. A decrease in the amounts of CP (except in T3), EE, CF, NDF, and ADF in the A. membranaceus straws treated with effective microorganisms was observed compared with that for the C and T1 (p<0.05). The NFE content of the straws increased with all treatments, except T1. Fermentation treatment subtly altered the fatty acid composition of A. membranaceus leaves and straws. In contrast, the calcium and vitamin E contents in A. membranaceus leaves and straws were increased after fermentation treatment (p<0.05). However, T3 yielded higher saponin content in straws compared to that by any of the other treatments (p<0.05). The effect of fermentation with microorganisms, maintained the low pH up to 48 h (p<0.05), whereas it was random for the straws. Therefore, the data suggest that fermentation treatment with microorganisms can improve digestion rate and have a positive effect on physiologically active substances and feed value.

가공조건 차이에 따른 한방차의 성분변화를 분석한 결과, 팽화공정 처리한 것은 볶음공정을 한 것보다 조회분, 수분, 조단백질, 고형분 용출율이 증가하였으며 조지방은 소폭 감소하였다. 벤조피렌 [B(α)P]함량은 0.35 ppb에서 0.18 ppb로 크게 감소하였다. 전체적으로 심한 열처리 과정이 없는데도 불구하고 B(α)P 이 검출된 이유는 식품 중 B(α)P 는 주로 음식을 조리, 가공할 때 식품의 주성분인 탄수화물, 단백질, 지방 등이 열분해 되어 생성되기 때문이다. 한방차에서 맛, 향, 색상 모두 큰 차이를 보이지 않으나 다소 텁텁한 느낌이 강하고, 시큼한 맛이 강하여 선호도를 떨어뜨리는 것으로 나타났다.

The following study is the result of herbal teas puffed at different temperatures between 140~220℃. Depending on treatment temperatures, the water contents decreased, while some carbonization occurred and crude ash contents relatively increased. Also, the crude protein and crude fat experienced little changes. B(α)P contents (0.16~0.17 ppb) showed little change according to treatment temperatures. From this result, the B(α)P content differed depending on the treatment temperature and raw materials. Solid elution rate figures of the herbal teas ranged from 0.27~0.45% (w/w) and the rate of solid elution increased along with higher puffing temperatures. The reason for the increase in solid elution rates is due to the breakage of cross bridges between the raw materials in the herbal tea which are carbohydrates, proteins, lipids and etc. after treatments of physical changes rather than chemical ones.

Roasting 온도를 80~140℃로 달리한 한방차의 성분 변화를 분석한 결과는 다음과 같다. 처리온도의 상승에 따라 수분함량이 감소하고, 일부 탄화가 발생하며 조회분 함량이 상대적으로 상승하는 소폭 변화가 있었고, 조단백질 및 조지방 함량은 거의 변화가 없는 것으로 나타났다. 벤조피렌 함량은 0.17~0.35ppb으로 처리온도가 상승할수록 B(α)P 함량이 증가하는 경향을 보였다. 이러한 결과를 볼 때, 처리온도와 원재료에 따라 B(α)P 함량에 차이가 발생한 것으로 나타났는데, roasting의 경우 실제 내부온도는 약 200℃내외에 그치지만 roaster 표면의 온도는 약 2,000℃에 이르기 때문에 이 표면과 직접 접촉한 부분에서 일부의 B(α)P가 생성된 것으로 판단된다. 한방차의 고형분용출율을 0.18~0.35%(w/w)를 나타내었는데, roasting 온도가 상승할수록 고형분용출율이 감소하는 경향을 나타내었다. 처리온도가 80~110℃처리구간에서는 큰 변화를 나타내지 않은 반면 110~140℃처리구간에서는 고형분용출율이 급격히 감소하는 경향을 보였다. 처리온도가 상승할수록 고형분용출율이 감소하는 것은 내부 조직이 치밀하여 상대적으로 용출이 어렵기 때문으로 생각된다.

본 연구는 식품소재로서의 활용가능성이 높고 생리활성 효과가 우수한 황기분말을 농도별로 첨가(0.5%, 1.0%, 1.5%, 2.0%)하여 어묵을 제조하고 품질특성을 측정하였다. 수분함량은 황기분말의 첨가여부에 관계없이 유의적인 차이는 나타나지 않았으며 pH는 전체적으로 6.75-6.86의 범위를 나타내었다. L값은 황기분말의 첨가량 증가에 따라 유의적인 차이는 나타나지 않았으나 a값은 높아지고 b값은 낮아지는 경향을 나타내었다. 황기분말 첨가 어묵의 절곡검사 결과는 모든 시료에서 AA로 측정되어 황기분말의 첨가가 어묵의 유연성에 영향을 미치지 않았으며 기계적 조직감 중 강도(strength)는 황기분말의 첨가량이 증가할수록 높아지는 경향을 나타내었고 검성(gumminess), 응집성(cohesiveness), 파쇄성(brittleness)은 황기분말 2.0% 첨가구가 가장 높게 측정되었다. 황기분말 첨가 어묵의 관능적특성 중 생선 비린내 정도(fishy smell)는 황기분말의 첨가량 증가에 따라 강도의 정도가 약해졌으며 구수한 냄새의 정도(savory smell)는 황기분말 1.0% 첨가구가 가장 높은 점수를 보였다. 구수한 맛의 정도(savory taste)는 황기분말 1.0% 첨가구가 6.27로 가장 높은 점수를 나타내었으며 느끼한 맛의 정도(oily taste)는 황기분말 첨가량의 증가에 따라 약해지는 것으로 평가되었다. 황기맛의 정도(Astragalus membranaceus taste)는 황기냄새의 정도와 마찬가지로 황기분말의 첨가량이 증가할수록 강하게 평가되었으며 관능적 조직감의 정도는 각 시료간 유의적인 차이가 나타나지 않았다. 관능적 선호도 평가결과, 황기분말 1% 첨가구가 냄새, 맛, 조직감, 전반적인 선호도 항목에서 가장 우수함을 나타내었다.

In this study, Astragalus membranaceus powder was added (3, 6, 9%) during yeast bread preparation and its effect on product quality was examined. The results showed that the dough pH increased as the Astragalus membranaceus powder content increased. However, dough volume during fermentation did not differ significantly among the samples. Bread volume decreased (p<0.001) with an increasing amount of Astragalus membranaceus powder. Also, as the Astragalus membranaceus powder content increased, the Hunter’s color ‘L’ value of the crust decreased and the ‘a’ value increased, and for the crumb, the ‘L’ value decreased and the ‘a’ value increased. Textural property analysis indicated that hardness increased with an increasing amount of Astragalus membranaceus powder. However, up to the 6% level, there were no sensory attribute differences among the samples.

Background: Astragalus membranaceus belonging to the Leguminosae family is often utilized as a traditional medicine. The aim of this study was to elucidate the basic breeding information required to develop short stem A. membranaceus cultivars. Methods and Results: Roots of A. membranaceus advanced yield trial (AYT) lines were harvested in late October 2017. Root yield of six AYT lines were increased in a range of 8.9 - 74.8% compared with ‘Aseong’ as control (check variety). The height of seven AYT lines were shorter than that of ‘Aseong’. In addition, stem diameter of nine AYT lines was thicker than that of ‘Aseong’. Consequently, 1502-56, 1503-90, and 1510-80 were selected as elite lines for the development of short stem cultivars. HPLC analysis was perfromed to identify lines with high level active components such as calycosin-7-O-β-D-glucoside and calycosin. The levels of both active components were higher in 1502-56, and 1503-90, but not in 1510-80 compared to ‘Aseong’. In addition, 2,2-Diphenyl- 1-picryl hydrazyl (DPPH) radical scavenging activity was higher in the 1502-56 compared to ‘Aseong’. Considering these results, two AYT lines, 1502-56 and 1503-90 were selected as short stem lines with high calycosin-7-O-β-D-glucoside and calycosin content. Conclusions: Taken together, Two short stem lines were identified in this study. In our future study, regional yield trial (RYT) will be conducted using these selected lines to develop new cultivars.

Background : The causative organism of Astragalus membranaceus (Hwanggi) root rot disease was known as Pythophthora drechsleri. In this study, a new species of root rot causative organism has been identified and reported. Methods and Results : Hwanggi, which has root rot disease, was collected in Jungsun-gun, Gangwon-do. The diseased root was cut into 1 ㎝ size and disinfected with 1% sodium hypochlorite solution for 1 minute and washed with sterile distilled water. The dried samples were dipped in the PDA medium and cultured in at 25℃ incubator. The 21 isolates were cultured under the same conditions and genomic DNA was extracted. The genomic DNA of isolated strains was extracted using a kit of Biofact. The isolated strains were identified using internal transcribed spacer (ITS) region sequencing. The ITS region was amplied using the primer pair ITS1 (5’-TCCGTAGGTGAACCTGCGG-3’) and ITS4 (5’-TCCTCCGCTTATTGATATGC-3’). The PCR was performed in a final volume of 50 ㎕ containing 10 mM Tris-HCl (pH 8.3), 50 mM KCl, 1.5 mM MgCl2, each dNTP at a concentration of 0.2 mM, 1.25 IU of Taq polymerase, each primer at a concentration of 10 pmol, and 2ul of the DNA template. Sequencing was performed by asking Macrogen. The analyzed sequences were compared using the Blast program provided by NCBI. As a result of identification of 21 strains, 13 Fusarium spp., 1 Peronospora sp. (causative organism of downy mildew), 3 Acremonium spp., 3 Uncultured fungus, 1 Phyrtophthora sp.. The main pathogen of wilt disease was known as Fusarium sp., a soil-borne pathogen. After analyzing the systematic taxonomic relationships, the systematic diagram was created using the neighbor-joining method. Conclusion : We have finally identified Phytophthora cryptogea as a causative organism of Hwanggi root rot disease and will check whether the disease has occurred on the inoculation through pot test.

Background : Exposure to Ultraviolet B (UVB) causes oxidative stress, inflammation, pigmentation and severe skin damage. Astragalus membranaceus (AG) has been used as a traditional medicine and have been studied various physiological activities. During the roasting process, bioactive substances is change including antioxidant substances. The aim is study the antioxidant effects and reactive oxygen species (ROS) inhibitory effect of the roasted A. membranaceus (R-AG). on Human dermal fibroblast (HDF) cells. Methods and Results : To prepare of R-AG samples, roasting machine was used. AG and R-AG were extracted to water and 70% ethanol. AG samples were evaluated the antioxidant potential by measuring the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2`-azino-bis 3-ethylbenzothiazoline-6-sulphonic acid (ABTS) scavenging activities. Additionally, total phenolic contents and total flavonoid contents was compared with antioxidant ingredients. AG and R-AGs were analyzed with HPLC determine the major compounds such as calycosin, mononetin and glycosides. The antioxidant activities of R-AG increased and changed in major compounds. In UVB exposed HDF cells, AGs did not affect cell viability and R-AG inhibited ROS more effectively than AG. Conclusion : From these results, R-AG can inhibit oxidative stress induced UVB in HDF cells.

Background : The isoflavonoids, triterpene and polysaccharides were know as major components in Astragalus membranaceus Bunge. Especially, isoflavonoids are classified as ononin, calycosin-glucoside of glycon binding and formononetin, calycosin of aglycone binding. In this study, we performed fermentation by a lot of microorganisms which were derived foods. Methods and Results : The A. membranaceus was extracted with 50% Ethanol and it was concentrated using rotary evaporator. In order to change in isoflavonoids, we fermented the A. membranaceus extracts with microorganisms which have β-glucosidase activity. we chose 20 microorganisms using esculin agar method and three strains with the highest activity of β -glucosidase were identified by pNP assay. The extracts of A. membranaceus were fermented during 3 days in sterilized distilled water, as 1 Brix and 3 Brix. The isoflavonoids of fermented extract, respectively days 0, 1, 2 and 3, were analyzed using High Pressure Liquid Chromatography (HPLC). In addition, the fermented A. membranaceus extracts was investigated about whitening effect using tyrosinase and radical scavenging activity by 1,1-diphenyl-2-picrylhydrazyl radical (DPPH). As a result of β-glucosidase activity by pNP assay, we selected two strains, Pediococcus pentosaceus (KACC 81017BP), Saccharomyces cerevisiae (KACC 83014BP). On the third day of fermentation of S. cerevisiae, isoflavone aglycone was the highest. The calycosin-glucoside and ononin in fermented A. membranaceus extract were reduced, calycosin and formononetin was increased compared to control. Also, we confirmed higher activity than control in tyrosinase inhibition rate on the third day of fermentation, as 78.38% and lower IC50 value of radical scavenging activity as 584.68 ㎍/㎖. Conclusion : From the above results, we may suggest that A. membranaceus aboveground parts might have useful as a safe material for functional food and pharmaceutics.

Background : Astragalus membranaceus belonging to the family of Leguminosae have been utilized as a traditional medicine. The aim of this study is to elucidate the basic information for breeding to superior Astragalus membranaceus cultivar. Methods and Results : Selection lines were developed by the medicinal crop breeding team of National Institute of Horticulture and Herbal Science (NIHHS), Rural Development Administration (RDA). The root of A. membranaceus lines were harvested in late October 2017. Calycosin-7-O-β-d-glucoside and calycosin were anlyzed by HPLC. The root yield of six selection lines were increased 8.9 - 74.8% compared with ‘Aseong’ (check variety). Also, the plant height of seven selection lines were shorter than ‘Aseong’. The calycosin–7-O-β -d-glucoside content was higher in six selection lines than ‘Aseong’. The calycosin content was higher in all selection lines except 1 line (1508 - 03) than that of ‘Aseong’. Conclusion : Three superior lines with short height and high yield were selected. These superior lines will conduct advanced yield trial to make varieties.

Astragalus membranaceus is a well known oriental medicinal herb. The polysaccharides of the aboveground parts (AMA) and the radix (AMR) of A. membranaceus are the most important functional constituents. Methods and Results: The aim of this study was to determine the effects of AMA and AMR on the oxidative damage induced in the skeletal muscle of rats subjected to exhaustive exercise. Sprague-Dawley rats were randomly divided into exercise and non-exercise groups; in the groups receiving the test compounds, AMA and AMR were administered orally for 30 days. Skeletal muscle samples were collected from each rat after running to exhaustion on a treadmill to determine the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) and the concentation of malondialdehyde (MDA). The antioxidant enzyme activities of SOD and GSH-Px of skeletal muscle of AMA- and AMR-treated groups were significantly higher than those of the control and commercial sports drink (SPD)-treated groups in exhaustive exercise rats. In addition, MDA concentrations in the skeletal muscle of the AMA- and AMR-treated groups were significantly lower than those of the control and SPD-treated groups. In the present study, the effects of AMA and AMR on exercise endurance capacity were also evaluated in mice subjected to a swimming exercise test. AMA and AMR supplementation prolonged the swimming time of mice and enhanced exercise endurance capacity. AMA and AMR possess the ability to retard and lower the production of blood lactate, and prevent the decrease of serum blood glucose. Conclusions: These results showed that, AMR and AMA exerted beneficial effect in mice, increasing the activity of the antioxidant systems and inhibiting oxidative stress induced by exhaustive exercise. The compounds improved exercise performance and showed anti-fatigue effects against exhaustive exercise.

Background : The major components in Astragalus membranaceus are isoflavonoids, triterpene and polysaccharides. Also, isoflavonoids was composed to aglycon and glucoside derivatives. In this study, we performed fermentation process (enzyme treatment) and steam processing (high temperature and pressure) to increase aglycon such as formononetin and calycosin. Methods and Results : The steam processing was performed at a lot of conditions, such as temperature (80, 100, 120℃) and time (30, 60 and 120 min). Fermentation processing carried out by A. membranaceus extract fermented with microorganisms which have high β -glucosidase activity (selection by esculin agar method) for detached glucose from isoflavonoids, converted to algycon. The isoflavonoids were analyzed using high pressure liquid chromatography (HPLC) in fermentation product of A. membranaceu extract by treated with steam processing at various conditions. As a result of β-glucosidase activity by pNP assay, we selected three strains, Pediococcus pentosaceus, Weissella cibaria and Saccharomyces cerevisiae. Isoflavone aglycon was the highest in fermentation product by S. cerevisiae for 3 days, but change of isoflavonoids was not observed in steam processing. The calycosin-glucoside and ononin were reduced in fermentation product of A. membranaceus extract, whereas calycosin and formononetin was increased. Conclusion : This results showed that isoflavone glycoside converted to isoflavone aglycon in A. membranaceus by fermentation process, it seems to be available for industrial use.

Background: Polysaccharides are the most important functional constituent in Astragalus membranaceus. The purpose of the present study was to evaluate the effect of polysaccharides isolated from the aboveground parts of A. membranaceus (AMA) and polysaccharides isolated from the roots of A. membranaceus (AMR) immune function by modulated cytotoxic T cell and Th1- and Th2- related cytokines kinetics. Methods and Results: Sprague-Dawley rats were randomly divided into exhaustive exercise case groups and non-exercise case, AMA and AMR samples were administered orally for 30 days (500 ㎎/㎏/day and 10 ㎎/㎏/day, respectively) and were compared to those rats in the groups fed commercial sports drink (SPD) and vehicle. Both exhaustive exercise groups and non-exercise groups had a lower ratio of CD4+ and CD8+ cells in the spleens of the rat fed AMA and AMR compared to those in the rats fed SPD and vehicle group. These results suggested that AMA and AMR promote an increase in the proportion of cytotoxic T cells. The IL-4- producing T lymphocytes decreased significantly in the AMR (10 ㎎/㎏/day) group compared to SPD and vehicle, whereas the AMA group increased the IL-4 concentration more than the SPD and vehicle in exhaustive exercise group. However, the populations of IFN-γ-producing T lymphocytes of AMR and AMA increased. AMA decreased the concentration of IFN-γ to inhibit the Th1 response and thereby increased the concentration of IL-4 to induce a Th2 response that was related to humoral immunity in the non-exercise group. Conclusions: These results showed that, in addition to Th1/Th2 regulation, AMR and AMA played an important immuno-modulatory role after exhaustive exercise-induced Th1/Th2 lymphocyte imbalance, which might be correlated with cytokine producing immunoregulatory cells.