• Archives of Pharmacal Research
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• v.27 no.2
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• pp.217-224
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• 2004

Previously, we reported that water-extracted Acanthopanax senticasus exhibited anti-meta-static activity by stimulating the immune system. In this study, we fractionated glycoproteins (EN-SP) from the soluble protein layer (GF-AS) of A. senticasus and determined their basic chemical properties. We also investigated the anti-tumor and immunostimulating activities of the fractionated glycoprotein, EN-SP. We found that intravenous (i.v.) administration of GF-AS dramatically inhibited metastasis of colon26-M3.1 carcinoma cells to the lung in a dose-dependent manner. In vitro analysis showed GF-AS to enhance the proliferation of splenocytes. GF-AS also stimulated peritoneal macrophage, which was followed by the production of various cytokines such as IL-1$\beta$, TNF-$\alpha$, IL-12 and IFN-${\gamma}$. Furthermore, the production of these cytokines was partially blocked when peritoneal macrophage was cultured with the polyclonal antibodies against GF-AS. The depletion of NK cells by rabbit anti-asialo GM1 serum partly abolished the inhibitory effect of GF-AS on lung metastasis of colon26-M3.1 cells. Using gel filtration, EN-SP, an active glycoprotein fraction, is isolated from GF-AS. While both GF-AS and EN-SP stimulated the proliferatation of splenocytes of normal mice, EN-SP showed higher anti-metastatic activity and more potently stimulated the proliferation of splenocytes compared to GF-AS. These results suggest the use of EN-SP, the fractionated glycoprotein from A. senticasus, can be used as a therapeutical reagent to prevent or inhibit tumor metastasis.

• The Korea Journal of Herbology
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• v.23 no.3
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• pp.141-147
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• 2008

Objectives : The cortex and root of Acanthopanax sessiliflorus (AR), a herbal medicine, have been used for several diseases including cancer in Oriental countries. Recently, we reported that AR has an immune-potentiating action. Oga-Power(OP) was made using extract from AR. For these reasons, we hypothesized that OP can potentiate the immune system in terms of accelerating proliferation rates of immune cells such as thymocytes and splenocytes. Methods : In this experiment, proliferation rates of thymocytes and splenocytes were measured using modified 3-[4,5-dimethy -lthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT). No production levels in macrophages isolated from normal mice were measured using Griess method. Results : In our results, treatment with OP accelerated proliferation rates of splenocytes, but did not affect those of thymocytes in vitro. On the other hand, proliferation rates of thymocytes was elevated in vivo. In addition, level of NO production from macrophage separated from abdominal cavity of normal mice was elevated by treatment with OP. Conclusions : In conclusion, OP has immune-potentiating action, by acceleration of splenocyte proliferation and elevation of NO production level from macrophages.

• Environmental Analysis Health and Toxicology
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• v.22 no.3
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• pp.263-269
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• 2007

Chitosan is a depolymerized and partially deacetylated derivative of chitin. We investigated the cytotoxicity of chitosan in cancer cell lines, such as P388, L1210, HCT-15, SK-HepG-1 and mouse splenocytes as a normal cell by MTT assay. To clarify whether chitosan enhances cytotoxicity of anticancer drugs, we also examined the cytotoxicity of combined treatment with chitosan and anticancer drugs, such as cisplatin, mitomycin C, and 5-fluorouracil in cancer cell lines in vitro. Chitosan ($37.5\;{\mu}g/mL,\;75\;{\mu}g/mL,\;112.5\;{\mu}g/mL,\;and\;150\;{\mu}g/mL$) showed concentration-dependent cytotoxicity in the cancer cell lines. In addition, chitosan showed relatively lower cytotoxicity in normal cells than in the cancer cell lines. Particularly, this trend was significant at high doses of chitosan, i.e. $112.5\;{\mu}g/mL,\;and\;150\;{\mu}g/mL$. Thus, these results suggest that chitosan may selectively induce the growth inhibition in cancer cell lines, compared to normal cells. Furthermore. the co-treatment of chitosan and anticancer drugs exhibited an apparant synergistic cytotoxicity in murine lymphoma cell lines, i.e. P388 and L1210 at $37.5\;{\mu}g/mL$ of chitosan rather than at $75\;{\mu}g/mL$ of chitosan, but such phenomenon could not be observed in solid tumor cell lines, i.e. HCT-15 and SK-HepG-1. However, chitosan did'nt reduced the cytotoxicity against normal mouse splenocytes induced by anticancer drugs. Therefore, it is concluded that the combination of chitosan and anticancer drugs might be useful for the cancer chemotherapy.

• Journal of Life Science
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• v.17 no.5 s.85
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• pp.694-698
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• 2007

Berberine, an alkaloid initially isolated from chinese herbal medicine, has antibiotic activities against a variety of organisms including bacteria, viruses, fungi, protozoans, and chlamydia. Furthermore, berberine has shown a number of beneficial effects, including anti-tumor, anti-inflammation, and vasodilatory effects. In this work we have investigated the effects of berberine on lymphocyte proliferation and GM-CSF production in mice. Mouse splenocytes were incubated with berberine and concanavalin A(Con A) to observe the effects on cell proliferation. The culture supernatants of splenocytes exposed to berberine, berberine plus LPS, and berberine plus Con A were harvested to assay GM-CSF. The cell proliferation of nice splenocytes exposed to berberine only($1{\mu}g/ml$) was increased significantly more than PBS(control) group. But the Con A-induced cell growth was inhibited by berberine. The GM-CSF production from mice splenocyte culture exposed to berberine only was increased in comparison with PBS(control) group, but the production of it with LPS or Con A was inhibited by berbeline. The present findings may explain lympocyte proliferating and regulating effects of berberine.

• Journal of Radiation Industry
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• v.7 no.2_3
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• pp.161-166
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• 2013

The purpose of this study was to investigate the cell proliferating and interleukin-2 producing activity of staphylococcal enterotoxin B by gamma-irradiation. Staphylococcal enterotoxin B was gamma-irradiated with the various doses of 0, 2, 20 and 50 kGy. SDS-PAGE analysis showed that gamma-irradiation caused the sharp decrease of the content of staphylococcal enterotoxin B and the effect was irradiating dose-dependent. Non-irradiated staphylococcal enterotoxin B increased the cell proliferation of splenocytes isolated from female Balb/c mouse, whereas 2 kGy-irradiated toxin significantly decreased the activity. 20 and 50 kGy-irradiated staphylococcal enterotoxin B was no effect. A similar effect on the interleukin-2 production of mouse splenocytes was observed with non-irradiated and irradiated staphylococcal enterotoxin B. It was considered to be due to the decrease of the antigenicity of staphylococcal enterotoxin B by gamma-irradiation. Therefore, these results suggest that gamma-irradiation can be effective for the decrease of the antigenicity of staphylococcal enterotoxin B as superantigen.

• Nutrition Research and Practice
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• v.17 no.2
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• pp.206-217
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• 2023

BACKGROUND/OBJECTIVES: The immunomodulatory effect of Platycodon grandiflorum (PG) has been reported, but studies on its mechanism are still lacking. This study was undertaken to confirm whether the hydrolyzed and fermented PG extract (HFPGE) obtained by adding hydrolysis and fermentation to the extraction process has an immune-enhancing effect in the in vivo system. MATERIALS/METHODS: Five-week-old BALB/c mice were divided into 4 groups: normal control group (NOR), control group (CON), 150 mg/kg body weight (BW)/day HFPGE-treated group (T150), and 300 mg/kg BW/day HFPGE-treated group (T300). The mice were administered HFPGE for 4 weeks and intraperitoneally injected with cyclophosphamide (CPA, 80 mg/kg BW/day) on day 6, 7, and 8, respectively, to induce immunosuppression. The levels of immunoglobulins (Igs) and cytokines were measured in the serum. In splenocytes, proliferation and cytokine levels were measured. RESULTS: Serum IgA, IgG, and IgM levels were observed to decrease after CPA treatment, which was recovered by HFPGE administration. The levels of serum interleukin (IL)-12, tumor necrosis factor (TNF)-α, IL-8, and transforming growth factor (TGF)-β were also decreased after exposure to CPA but increased after HFPGE administration. Decreased splenocyte proliferation was seen in CPA-treated mice, but was observed to increase in the T150 and T300 groups as compared to the NOR group. Compared to the CON group, splenocyte proliferation stimulated with concanavalin A (ConA) or lipopolysaccharide (LPS) in the HFPGE-treated groups was significantly increased. The cytokines secreted by ConA-stimulated splenocytes (IL-2, IL-12, interferon-γ, TNF-α) were increased in the T150 and T300 groups, and cytokines secreted by LPS-stimulated splenocytes (IL-4, IL-8, TGF-β) were also increased by HFPGE administration. CONCLUSION: These results suggest that HFPGE stimulates the immunity in immunosuppressed conditions, thereby enhancing the immune response. Therefore, it is expected that HFPGE has the potential to be used as functional food and medicine for immune recovery in various immunocompromised situations.

• Journal of the Korean Applied Science and Technology
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• v.40 no.5
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• pp.955-964
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• 2023

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.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.3 no.1
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• pp.129-147
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• 1997

Radiotherapy is an irreplaceable method of cancer treatment. But it has several side effects, especially damages to the hemopoietic and Immune system. Therefore radioprotectors are required to treat cancer successfully. A lot of Herbs and Herbal prescriptions are reported to have radioprotective effects. Above all, those to support the healthy energy and strengthen the body resistance are found more effective. This study was performed to evaluate the radioprotective effects of prescription Shenrong Fuzheng Tang(S.F.T.), which consists of 16 kinds of herbs. We investigated proliferation of murine splenocytes, secretion of colony-stimulating-factors(CSFs), immunocompetence after irradiation in-vitro, and Endogenous spleen colony assay, survival assay in-vivo. When splenocytes were cultured with Shenrong Fuzheng $Tang(S.F.T.)(500{\mu}g/ml)$, proliferation was enhanced 5.7 times compared to control cultured with medium alone(p<0.05) and, showed highest proliferation at 4th day after incubation. In order to evaluate stimulation of hemopoiesis of Shenrong Fuzheng Tang(S.F.T.), the supernatant of splenocytes cultured with optimal concentration of Shenrong Fuzheng Tang(S.F.T.) was used to measure CSFs secretion. The result showed enhanced secretion of colony-stimulating-factors (CSFs) compared to control(p<0.05). To evaluate the protective effect of lymphocytes from irradiation, proliferation of splenocytes stimulated by LPS and ConA after incubation with Shenrong Fuzheng Tang(S.F.T.) for 24h Prior to Irradiation$(1{\sim}3\;Gy)$ was measured. The results showed higher proliferation of Shenrong Fuzheng Tang(S.F.T.) treated cells than that of non-treated cells. And percentage increases of irradiated splenocytes per non-irradiated splenocytes were also higher in Shenrong Fuzheng Tang (S.F.T.)-treated cells than control. Endogenous spleen colony assay. to evaluate the protection of hemopoietic cells from irradiation, showed increased number of colonies(p=0.03) in Shenrong Fuzheng Tang(S.F.T.) treated murine spleen$(10.3{\pm}1.9)$ compared to non-treated murine spleen$(3.4{\pm}0.8)$. Survival time of mice irradiated with lethal dose of ${\gamma}-ray(9Gy)$ was prolonged in Shenrong Fuzheng Tang(S.F.T.) treated group prior to irradiation as compared to non-treated group. According to these results we can suggest that prescription Shenrong Fuzheng Tang(S.F.T.) has radioprotective effects and can be used to protect the hemopoietic and immune system from damages of anti-cancer radiotherapy.

• Journal of Animal Science and Technology
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• v.49 no.2
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• pp.213-224
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• 2007

In this study, hatched male broiler chicks(Ross) were fed on a basal diet and LPS was administered via intraperitoneal injection three times every other day, on the 9th, 11th and 13th days of the experiment, and then PBMC and splenocytes were isolated on day 14. The degree of alama blue reduction was evaluated at 4, 24, 48, 96 and 120 h in the splenocytes, and at 4, 8, 12, 24 and 48 h for PBMC of incubation after the addition of alama blue solution to the media. The cell numbers used in this experiment were 103, 104 and 105 cells per well, and the con A levels were 0.0, 1.0, 5.0, and 10.0 ㎍ per ml of medium. 1. The degree of alama blue reduction was found to increase in a linear fashion with increasing incubation time and cell numbers, for both splenocytes and PBMC. 2. During acute phase response, the degree to which alama blue was reduced was significantly elevated (p<0.05) at an incubation time of 24 hr for the splenocytes, 4 hr for PBMC, and a cell number of 105 cells per well, respectively. 3. The raised reduction of alama blue to control was linear with Con A levels in medium, and higher reduction in Con A 10.0 ㎍ relative to 1.0 or 5.0 ㎍ in ml medium was shown 4. The medium with autologous serum evidenced a significantly (p<0.05) higher reduction of alama blue relative to FBS. 5. Splenocytes and PBMC from the LPS-injected birds evidenced significantly higher levels of alama blue reduction regardless of incubation time, number of cells, level of Con A added, or serum type, as compared with what was observed in normal birds. The results indicated that the assay conditions for proliferative activity using the alama blue method in birds in which the acute phase response had been activated via intraperitoneal LPS injection requires 4 hrs of incubation for PBMC, 24 hrs of incubation for splenocytes, and 10㎍ of Con A per ml of medium.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.2
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• pp.167-175
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• 2005

Houttuynia cordata THNUB (He; Uh-Sung-Cho) is a medicinal plant which has been widely used as a component of blood-building decoctions. This study was performed to investigate the immunomodulative effect of He in mice. In vitro experiment, the mice splenocytes proliferation and three kinds of cytokines (IL-1$\beta$, IL-6, TNF- $\alpha$) production by mice peritoneal macrophages cultured with six (methanol, hexane, chloroform, ethylacetate, butanol and water) fractions of He were used to indicate the immunomodulative effect. Ex vivo experiment, the different concentrations of He water extract was orally administrated every other day for two weeks. The production of cytokines IL-1$\beta$, IL-6, TNF- $\alpha$) secreted by activated macrophages and the mice splenocytes proliferation were used as an index for the immunocompetence. The supplementation of all six fractions of He enhanced the splenocytes proliferation at the level of 6.58$\pm$1.23∼47.82$\pm$5.48 compared to that of control in the range of 1∼50 $\mu\textrm{g}$/mL. IL-1$\beta$ production was significantly increased with the supplementation of chloroform and water extract of He. Higher level of IL-6 production was detected by the supplementation of ethylacetate, butanol and water extract. TNF - $\alpha$ production was enhanced by the supplementation of all six fractions of He. From the ex vivo study, the highest proliferation of splenocytes was seen from the mice orally administrated with the He water extract at the concentration of 500 mg/kg bw In case of cytokines production, IL-1$\beta$, IL-6, and TNF- $\alpha$ release by activated peritoneal macrophages were augmented by the oral administration of He water extract. These results indicated that He may enhance the immune function by regulating the splenocytes proliferation and cytokines production capacity in mice.