괭생이 모자반 추출물의 비장세포 면역활성 증강 효과

김동섭,성낙윤,한인준,이병수,박상윤,노은영,엄지,김건,김경아 한국영양학회 2019 Journal of Nutrition and Health Vol.52 No.6

Purpose: This study examined the immunological activity and optimized the mixture conditions of Sargassum horneri (S. horneri) extracts in vitro and in vivo models. Methods: S. horneri was extracted using three different methods: hot water extraction (HWE), 50% ethanol extraction (EE), and supercritical fluid extraction (SFE). Splenocyte proliferation and cytokine production (Interleukin-2 and Interferon-γ) were measured using a WST-1 assay and enzyme-linked immunosorbent assay, respectively. The levels of nitric oxide and T cell activation production were measured using a Griess assay and flow cytometry, respectively. The natural killer (NK) cell activity was determined using an EZ-LDH kit. Results: Among the three different types of extracts, HWE showed the highest levels of splenocyte proliferation and cytokine production in vitro. In the animal model, three different types of extracts were administrated for 14 days (once/day) at 50 and 100 mg/kg body weight. HWE and SFE showed a high level of splenocyte proliferation and cytokine production in the with and without mitogen-treated groups, whereas EE administration did not induce the splenocyte activation. When RAW264.7 macrophage cells were treated with different mixtures (HWE with 5, 10, 15, 20% of SFE) to determine the optimal mixture ratio of HWE and SFE, the levels of nitric oxide and cytokine production increased strongly in the HWE with 5% and 10% of SFE containing group. In the animal model, HWE with 5% and 10% of SFE mixture administration increased the levels of splenocyte proliferation, cytokine production, and activated CD4 + cell population significantly, with the highest level observed in the HWE with 5% of SFE group. Moreover, the NK cell activity was increased significantly in the HWE with 5% of SFE mixture-treated group compared to the control group. Conclusion: The optimal mixture condition of S. horneri with immune-enhancing activity is the HWE with 5% of SFE mixture. These results confirmed that the extracts of S. horneri and its mixtures are potential candidate materials for immune enhancement.

Ulmus Macrocarpa Water Extract Prolongs Splenocyte Life Span

Kyung-Hwa Kang(강경화),Sook Kyung Hyun(현숙경),Hye Jin Hwang(황혜진),Byoung Woo Kim(김병우),Cheol Min Kim(김철민),Kyung Tae Chung(정경태),Jong-Hwan Lee(이종환) 한국생명과학회 2015 생명과학회지 Vol.25 No.10

Ulmus macrocarpa 자양강장 및 생리활성 물질로 이용되어 왔다. U, macrocarpa 열수 추출물(UMWE)이 일반적인 세포배양 조건에서 비장세포 수명연장에 미치는 효과에 대한 연구를 진행하였다. 100 μg/ml UMWE를 비장세포에 처리하여 실험을 진행하였다. 살아있는 세포확인은 Hoechst 33342 염색법과 세포생존관련 인자의 변화는 Western blot으로 확인하였다. 사이토카인 변화는 ELISA로 검증하였다. UMWE는 비장세포에 대하여 향상된 세포 생존력을 보였다. UMWE를 48시간과 96시간째 처리된 비장세포의 PI3K 및 ERK1/2의 인산화를 증가시켰다. 더욱이, 48시간과 96시간때에 Bcl-2의 발현량도 증가하였다. 반면, UMWE는 48시간과 96시간에 caspase-3의 활성이 줄어들었다. ICAD 단백질은 48시간에 증가하였다. UMWE는 조혈 및 세포생존력에 영향을 미치는 IL-2 cytokine량은 줄었지만 반면, IL-4 hematopoietin cytokine의 양은 증가하였다. UMWE는 48시간과 96시간에 증가된 IFN-γ level을 나타내었고 IL-12의 경우는 증가패턴을 보이는 효과를 발휘하였다. 이러한 결과는 UMWE가 다양한 신호전달 및 사이토카인 조절을 통해 비장세포 수명연장을 할 수 있다는 것으로 사료된다. Ulmus macrocarpa has been used in Korean medicinal food material to physical disorder or tonic material. The purpose of the present study was to evaluate splenocyte life span expansion effects of Ulmus macrocarpa water extract (UMWE) in general cell culture condition. Splenocytes were handled in the presence of 100 μg/ml UMWE for several different time conditions. Live cells were detected with Hoechst 33,342 dye and cell survival molecules were identified through Western blot. Changes in level of cytokine synthesis were evaluated by ELISA. UMWE showed an effect on increased splenocyte survival. UMWE elevated slightly PI3K phosphorylation and ERK1/2 phosphorylation used at 48 hr and 96 hr. Moreover, Bcl-2 was elevated at 48 hr and 96 hr in UMWE-treated splenocytes. UMWE decreased caspase-3 level at 48 hr and 96 hr. ICAD protein increased at 48 hr culturing time. Hematopoietin IL-2 cytokine was down-regulated, however IL-4 hematopoietin cytokine was up-regulated in UMWE treated cell culture media. Increased IFN-γ levels were verified in the supernatant of UMWEtreated cells in all periods (48 hr and 96 hr). Increased patterns in the production of IL-12 cytokine occurred as compared with control after 48 and 96 hr in UMWE-treated-cell cultures. These results suggested that UMWE can prolong splenocyte life span by controlling various signal factors and cytokines.

젖소 초유로부터 분리한 Insulin-like Growth Factor-1 분획이 In Vitro에서 마우스의 면역 활성에 미치는 영향

황경아,양희진,이수원,Hwang Hyung-A,Yang Hee-Jin,Lee Soo-Won 한국축산식품학회 2006 한국축산식품학회지 Vol.26 No.1

젖소 초유 whey 내에 존재하는 IGF-I을 분리하기 위하여 ultrafiltration을 사용하여 30과 1 kDa 사이의 IGF-I rich fraction을 분획하였다. 분획한 IGF-I rich fraction내에 IGF-I의 존재 여부는 SDS-PACE와 western blotting으로 확인하였고, sandwich ELISA로써 분획 내 IGF-I을 정량하였다. 그 결과 IGF-I은 단백질 mg당 10 ng으로 측정되었다. 분획한 IGF-I rich fraction을 마우스 복강 macrophge의 면역 활성에 미치는 영향 즉, IL-6, NO, $TNF-{\alpha}$, Phagocytosis, $H_{2}O_{2}$,의 분비 유도 및 생성량을 측정하였고, splenocyte의 면역 활성으로는 splenocyte의 증식 효과와 natural killer cell의 항암작용을 측정하였다. 실험 결과 IGF-I rich fraction $1{\mu}g/mL$ 투여군에서 IL-6의 생성량은 9.85 ng, NO의 생성량은 $17.1{\mu}M$, phagocytosis는 양성 대조구에 대비하여 78.3%의 탐식작용을 나타내었고 $TNF-{\alpha}$와 $H_{2}O_{2}$의 생성량은 대조구에 대비하여 각각 34.5 및 6% 더 많은 양을 생산하였다. Splenocyte 면역활성에 미치는 영향으로 T cell과 B cell 증식은 대조군에 대비하여 각각 36 및 76%로 더 높은 증식 효과를 나타내었고, NK cell의 활성은 대조군보다 55.4%의 높은 활성을 보였다. Insulin-like growth factor-I(IGF-I) rich fraction, which was obtained molecules ranged between 30 and 1 kDa, was fractionated by ultrafiltration from bovine colostral whey. IGF-I rich fraction was confirmed by SDS-PACE and western blotting and then the quantity of IGF-I was measured by sandwich ELISA. ICF-I concentration in IGF-I rich fraction was 10 ng/mg proteins. IGF-I rich fraction, standard IGF-I and colostral whey weie treated to murine peritoneal macrophages. And then we experimented that effect of immune activity on macrophage and splenocyte. As a result, in group treated with IGF-I rich fraction $1{mu}g/mL$, production of interleukin-6 and nitric oxide were 9.85 ng and $17.17{\mu}M$ and production of phagocytosis, tumor necrosis factor-${\alpha}\;and\;H_{2}O_{2}$ were 78.3, 34.5 and 6% compared to the control group. In splenocyte immune response, B cell and T cell proliferation and NK cell activity were 103, 126 and 22.2% in group treated with IGF-I rich fraction $1{\mu}g/mL$ to compared to the control, respectively.

Immuno-regulatory Effect of Dryopteriscrassirhizoma Extract on Chicken Splenocyte

Baatartsogt. O,Urantulkhuur. B,S.W. Suh,H.K. Lim,Y.K. Kim,K.O.Park,J.H. Kim,S.H. Kim,P. K. mandal,K. D. Choi 한국가금학회 2008 한국가금학회 정기총회 및 학술발표회 Vol.25 No.-

We investigated the effect of herbal extract of Dryopteris crassirhizoma on the cell viability, immune response, and nitric oxide production of chicken splenocyte. Nitrite levels were assayed in the culture supernatant of splenocyte after 24h by the Griess method. The expressions of pro-inflammatory cytokines such as interleukin (IL-2, IL-6, IL-8, IL-18) and the housekeeping gene GAPDH were studied using RT-PCR. Splenocyte were cultured with the herbal extracts at 37℃ in 96-well plate for 24h. Not cytotoxic effect was observed at 20 ㎍ and 40 ㎍. The NO production increased dose dependently with the treatment of herbal extract. which suggest that this herbal extract has no anti-inflammatory effect. In cytokine expression it was observed that the herbal extract induced the expression of IL-2 and suppressed the expression of IL-6 and IL-18, however there was no effect on the expression of IL-8. It may be concluded that the Dryopteris crassirhizoma extract is non toxic and have no anti-inflammatory effect and it has immune stimulation effect through IL-2.

Broussonetia kazinoki Siebold stimulates immune response in ovalbumin-immunized mice

Jung, Da-Young,Ha, Hye-Kyung,Lee, Ho-Young,Lee, Jin-Ah,Jeong, Seung-Il,Choi, Young-Jae,Shin, Hyeun-Kyoo The Society of Korean Medicine 2011 대한한의학회지 Vol.32 No.3

Objective: To evaluate the immune-stimulatory potential of extracts of Broussonetia kazinoki Siebold (BK) on specific cellular and humoral immune responses in ovalbumin (OVA)-immunized mice. Material and Methods: C57BL/6 mice were immunized intraperitoneally with OVA/alum ($100{\mu}g/200{\mu}g$) on days 1, 8, and 15. BK (100, 300 or 1000 mg/kg) was given to mice orally for 21 days (from day 1 to day 21). At day 22, OVA-, lipopolysaccharide (LPS)- and concanavalin A (Con A)-stimulated splenocyte proliferation and OVA-specific and total antibodies were measured in plasma. Further, the effects of BK on expression of cytokine mRNA in OVA-immunized mice splenocytes were evaluated by RT-PCR analysis. Results: BK significantly enhanced OVA-, LPS-, and Con A-induced splenocyte proliferation in OVA-immunized mice (p<0.01). BK also significantly enhanced total IgM and OVA-specific IgG1 levels in plasma compared with the OVA control group. Moreover, BK up-regulated significantly the expression of mRNA level of IL-2 and IFN-${\gamma}$ in splenocytes. Conclusions: BK has immune-stimulating activity in an OVA-immunized mouse model system, enhancing the Th1 immune response. BK showed no cytotoxicity in this system, suggesting that BK may be a safe and effective adjuvant in humans.

Broussonetia kazinoki Siebold stimulates immune response in ovalbumin-immunized mice

Da Young Jung,Hyekyung Ha,Hoyoung Lee,Jin-Ah Lee,Seung-Il Jeong,Young-Jae Choi,Hyeun-Kyoo Shin 대한한의학회 2011 대한한의학회지 Vol.32 No.3

Age-Related Effects of Sodium Arsenite on Splenocyte Proliferation and Th1/Th2 Cytokine Production

Yuri Cho,김대경,Kyong Hoon Ahn,백문정,최종민,Jung Eun Ji,Jong Hoon Won,Zhicheng Fu,Ji Min Jang 대한약학회 2012 Archives of Pharmacal Research Vol.35 No.2

Aging is associated with immune dysfunction and conditions such as inflamm-aging and immunosuppression. Arsenic, an environmental contaminant distributed worldwide, affects the immune system. This study tested the hypothesis that arsenic has distinct effects on T cell proliferation and the production of cytokines by activated T cells. Murine splenocytes from young (2 months) and aged (24-26 months) C57BL/6 mice were exposed to arsenite (As3+), the most toxic form of inorganic arsenic, and stimulated with concanavalin A (Con A) or anti-CD3 antibody. T cell proliferation decreased significantly in response to Con A and anti-CD3 at subtoxic doses of arsenite in splenocytes from both young and aged mice. Arsenite, added concurrently with Con A or anti-CD3, significantly inhibited the production of interleukin-2 (IL-2), interferon-γ (IFN-γ), and interleukin-4 (IL-4) by splenocytes from young mice and significantly reduced the production of IL-10 by splenocytes from aged mice. In contrast, the production of IL-2 and IL-4 by splenocytes from aged mice was only slightly affected by arsenite. The results show that arsenic exposure reduces the immune response in splenocytes. Moreover,this effect may be influenced by aging.

홍삼의 아미노당(Arginine-Fructose-Glucose) 강화농축액의 면역증진 효과

현선희(Sun Hee Hyun),김영숙(Young Sook Kim),이종원(Jong Won Lee),한창균(Chang-Kyun Han),박민선(Park Min Seon),소승호(Seung-Ho So) 한국식품영양과학회 2018 한국식품영양과학회지 Vol.47 No.1

본 실험은 홍삼의 증숙(열처리)과정에서 다량 생성되는 것으로 알려진 아미노당(arginine-fructose-glucose, AFG)을 강화한 농축액의 면역증강 효과를 확인하고자 실시하였다. In vitro에서 정상 마우스의 비장세포를 이용하여 AFG 강화농축액이 비장세포 증식능에 미치는 영향을 알아본 결과, AFG 강화농축액은 비장세포 증식을 증가시키고 LPS와 Con A로 유도되는 비장세포 증식도 유의하게 증가시켰다. Ex vivo에서 수컷 Balb/c 마우스에 AFG 강화농축액을 250, 500, 1,000 mg/kg 용량으로 매일 1회씩 10일 동안 경구투여 하면서 CY로 면역을 저하시킨 후, SRBCs로 감작하여 면역을 유도하여 면역장기인 비장과 흉선의 무게와 항체생성능을 측정하였다. AFG 강화농축액은 CY에 의해 감소하는 비장과 흉선의 무게를 회복하였고, 특히 흉선의 무게를 유의성 있게 증가시켰다. 또한, 비장세포수와 비장세포중의 IgM 항체생성 세포수도 용량 의존적으로 증가하였으며, 특히 500, 1,000 mg/kg 투여군은 현저하게 증가하였다. 이상의 실험 결과 AFG 강화농축액은 비장세포 증식을 촉진하고 CY에 의해 저하된 체액성 면역반응을 활성화시켜 면역기능을 향상시킬 것으로 생각된다. The purpose of this study was to investigate the immuno-modulatory effects of arginine-fructose-glucose (AFG)-enriched extracts of red ginseng. The effect on proliferation in normal mouse splenocytes was measured in vitro. When mouse splenocytes were exposed to various concentrations of AFG-enriched extracts with mitogens (concanavalin A, lipopolysaccharide), splenocyte proliferation significantly increased. In vivo, AFG-enriched extracts were orally administered (250, 500, and 1,000 mg/kg/d for 10 days) with cyclophosphamide (CY, 50 mg/kg, i.p.) to male balb/c mice, which were sensitized with sheep red blood cells (SRBCs) to measure the number of IgM antibody-forming cells in splenocytes. AFG-enriched extracts restored the weights of the spleen and thymus, which were reduced by CY. AFC counts per million splenocytes and total splenocyte number significantly increased in a dose-dependent manner compared with the CY Control group. These results indicate that AFG-enriched extracts increased humoral immunity against CY-induced immunosuppression in animal models.

The effect of bee pollen and its flavonoids on immune-modulating in mice

박지아,정일경,최윤식 한국응용과학기술학회 2023 한국응용과학기술학회지 Vol.40 No.5

Bee pollen is a valuable apitherapeutic product and has been known to have diverse biological activities, including antimicrobial, anti-inflammatory, and even anticancer activity. However, its effect on the immune system is not well studied and is rather controversial. This study intended to elucidate the biological activity of bee pollen on immunity. For this purpose, we used lyophilized bee pollen after wet grinding, which shows increased extraction of bioactive components and enhanced biological activity. First, lyophilized bee pollen after wet grinding significantly increased the proliferation of splenocytes isolated from normal mice. On the other hand, lyophilized bee pollen after wet grinding dose-dependently reversed splenocyte proliferation by concanavalin A or lipopolysaccharide. To clarify the activity of bee pollen on immunity lyophilized bee pollen after wet grinding was administered daily to mice for five weeks and isolated splenocytes. In this study, there was no significant difference in the population of immune cells and the size of spleen between bee pollen- and sterile water-treated groups. However, proliferation of splenocyte isolated from bee pollen-administered animals was boosted by both concanavalin A and lipopolysaccharide. Finally, kaempferol, a well-known flavonoid from bee pollen, dose-dependently increased splenocyte proliferation by both Con A and LPS. On the other hand, naringenin, another flavonoid in the bee pollen, dose-dependently inhibited the proliferation of splenocytes by Con A and LPS. Together, these data indicate that bee pollen may be able to prime the immunity to boost immune reaction after inflammation.

Bioactive Compounds / Food Microbiology : Dead Lactobacillus plantarum Stimulates and Skews Immune Responses toward T helper 1 and 17 Polarizations in RAW 264.7 Cells and Mouse Splenocytes

( Hyun Ah Lee ),( Hyunung Kim ),( Kwang Won Lee ),( Kun Young Park ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.3

This study was undertaken to evaluate the immunomodulatory effect of dead nano-sized Lactobacillus plantarum (nLp) in RAW 264.7 cells and murine primary splenocytes. nLp is a dead, shrunken, processed form of L. plantarum nF1 isolated from kimchi (a traditional Korean fermented cabbage) and is less than 1 μm in size. It was found that nLp treatment stimulated nitric oxide (NO) production more in RAW 264.7 macrophages than pure live L. plantarum (pLp), and that the stimulatory properties were probably largely derived from its cell wall. In addition, nLp induced murine splenocyte proliferation more so than pLp; in particular, a high dose of nLp (1.0 × 10(11) CFU/ml) stimulated proliferation as much as lipopolysaccharide at 2 μg/ml. Moreover, according to our cytokine profile results in splenocytes, nLp treatment promoted Th1 (TNF-α, IL-12 p70) responses rather than Th2 (IL-4, IL-5) responses and also increased Th17 (IL-6, IL-17A) responses. Thus, nLp stimulated NO release in RAW 264.7 cells and induced splenocyte proliferation more so than pLp and stimulated Th1 and Th17 cytokine production. These findings suggested that dead nLp has potential use as a functional food ingredient to improve the immune response, and especially as a means of inducing Th1/Th17 immune responses.