The effects of agmatine on the apoptosis of immune cells in septic rats / 中华急诊医学杂志

Objective:By establishing the rats model of sepsis induced by endotoxin, exploring the effects of agmatine on the apoptosis of splenocyte and dendritic cells in the septic rats.Methods:SD rats ( healthy and clean, male, 90) were randomly(random number) divided into control groups, endotoxin groups and agmatine groups, 30 rats per group. The control groups were injected with normal saline via femoral vein (10 mL/kg), endotoxin groups were injected with lipopolysaccharide via femoral vein (10 mg/kg), agmatine groups were injected with lipopolysaccharide via femoral vein (10 mL/kg) and intraperitoneal injected of agmatine (200 mg/kg).The three groups were randomly selected 10 rats separately at 0 h, 12 h, 24 h (marked as 0 h, 12 h, 24 h subgroups, n=10) , anesthetized to death. Weigh the spleen; HE staining was used to observe the pathological changes of the spleen; The cell apoptosis of splenocyte and dendritic cells were detected by flow cytometry. The results were statistically analyzed using SPSS 17.0 software, analyzed by independent sample t test, P<0.05 was considered statistically significant. Results:The weights (g) of spleen in 12 h (1.2633±0.0652) , 24 h (1.5576±0.0711) subgroups of the endotoxin groups were increased significantly than the corresponding subgroups[(0.8876±0.0361),(0.9079±0.0425)]of the control groups ( P<0.05) , the 12 h (1.1052±0.0585) , 24 h (1.3262±0.0682) subgroups of the agmatine groups were decreased significantly than the corresponding subgroups of the endotoxin groups ( P<0.05) . HE staining showed that the spleen tissue inflammatory reaction in the endotoxin groups were worse than the control group ( P<0.05) , the agmatine groups were better than the endotoxin group ( P<0.05) . The apoptosis rates of the splenocyte[(13.31±1.26)、(19.53±1.68)]and dendritic cells[(19.5±1.52)、(16.09±1.15)]in the spleen from 12 h, 24 h subgroups of the endotoxin groups were significantly higher than the corresponding subgroups[(6.27±0.71),(6.01±0.67) and (4.99±0.51)、(5.30±0.66)]of the control groups ( P<0.05) , the 12 h[(9.19±0.95),(12.19±1.25)], 24 h [(12.71±1.19),(10.76±1.09)subgroups of the agmatine groups were significantly lower than the corresponding subgroups of the endotoxin groups ( P<0.05) ; The apoptosis rates of the splenocyte in the 24 h subgroups of the endotoxin groups and the agmatine groups were significantly higher than the 12 h subgroups of the same group ( P<0.05) , but the apoptosis rates of the dendritic cells were significantly lower than the 12h subgroups of the same group ( P<0.05) . Conclusion:The apoptosis of splenocyte and dendritic cells were closely related to the occurrence and development of sepsis, Agmatine could inhibit the apoptosis of splenocyte and dendritic cells the rats with sepsis.

Splenocyte-mediated immune enhancing activity of Sargassum horneri extracts / 한국영양학회지

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.

Brown Adipocyte and Splenocyte Co-Culture Maintains Regulatory T Cell Subset in Intermittent Hypobaric Conditions

BACKGROUND: Brown adipocytes have thermogenic characteristics in neonates and elicit anti-inflammatory responses. We postulated that thermogenic brown adipocytes produce distinctive intercellular effects in a hypobaric state. The purpose of this study is to analyze the correlation between brown adipocyte and regulatory T cell (T(reg)) expression under intermittent hypobaric conditions. METHODS: Brown and white adipocytes were harvested from the interscapular and flank areas of C57BL6 mice, respectively. Adipocytes were cultured with syngeneic splenocytes after isolation and differentiation. Intermittent hypobaric conditions were generated using cyclic negative pressure application for 48 h in both groups of adipocytes. Expression levels of T(regs) (CD4 + CD25 + Foxp3 + T cells), cytokines [tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10), and the programmed death-ligand 1 (PD-L1)] co-inhibitory ligand were examined. RESULTS: Splenocytes, cultured with brown and white adipocytes, exhibited comparable T(reg) expression in a normobaric state. Under hypobaric conditions, brown adipocytes maintained a subset of T(regs). However, a decrease in T(regs) was found in the white adipocyte group. TNF-α levels increased in both groups under hypobaric conditions. In the brown adipocyte group, anti-inflammatory IL-10 expression increased significantly; meanwhile, IL-10 expression decreased in the white adipocyte group. PD-L1 levels increased more significantly in brown adipocytes than in white adipocytes under hypobaric conditions. CONCLUSION: Both brown and white adipocytes support T(reg) expression when they are cultured with splenocytes. Of note, brown adipocytes maintained T(reg) expression in intermittent hypobaric conditions. Anti-inflammatory cytokines and co-inhibitory ligands mediate the immunomodulatory effects of brown adipocytes under altered atmospheric conditions. Brown adipocytes showed the feasibility as a source of adjustment in physical stresses.

Nano chloroquine delivery against Plasmodium berghei NK65 induced programmed cell death in spleen

Objective: To compare the protective effects of chitosan-trypolyphosphate (CS-TPP) nanoparticle conjugated chloroquine(CQ) with effect of CQ alone on the reversal of splenic damages and induction of apoptosis. Methods: Different researches have been carried out to explore the potential role of chitosan based drug delivery system against parasitic diseases. After successive Plasmodium berghei NK65 parasiste infection by intraperitoneal injection in Swiss mice and subsequent parasite development, the ROS generation, anti-apoptotic and pro apoptotic protein levels in spleen were measured. To analyze caspases, flow cytometry study was performed with annexin V-FITC and with PI staining. Results: The results revealed that ROS mediated caspase 3 and 9 activation and the induction of apoptosis occurred during the parasitic infection. However, CS-TPP conjugated CQ was relatively better in reversing the splenic damage compared with similar effects of CQ alone. Conclusions: This study indicates that Plasmodium berghei NK65 induces apoptosis in the spleen. The study further shows that CS-TPP nanoparticles conjugation with CQ have positive influence on the recovery of damaged host's system towards maintenance of normal homeostasis, and this is shown to be selective to CS-TPP conjugated CQ treated animals only.

Nano chloroquine delivery against Plasmodium berghei NK65 induced programmed cell death in spleen

Objective:To compare the protective effects of chitosan-trypolyphosphate (CS-TPP) nanoparticle conjugated chloroquine(CQ) with effect of CQ alone on the reversal of splenic damages and induction of apoptosis.Methods:Different researches have been carried out to explore the potential role of chitosan based drug delivery system against parasitic diseases. After successive Plasmodium berghei NK65 parasiste infection by intraperitoneal injection in Swiss mice and subsequent parasite development, the ROS generation, anti-apoptotic and pro apoptotic protein levels in spleen were measured. To analyze caspases, flow cytometry study was performed with annexin V -FITC and with PI staining.Results:The results revealed that ROS mediated caspase 3 and 9 activation and the induction of apoptosis occurred during the parasitic infection. However, CS-TPP conjugated CQ was relatively better in reversing the splenic damage compared with similar effects of CQ alone.Conclusions:This study indicates that Plasmodium berghei NK65 induces apoptosis in the spleen. The study further shows that CS-TPP nanoparticles conjugation with CQ have positive influence on the recovery of damaged host’s system towards maintenance of normal homeostasis, and this is shown to be selective to CS-TPP conjugated CQ treated animals only.

Effect of Immunos on immune response cells by azathioprine Induced immunosuppression in mice / Монголын Анагаах Ухаан

Background@#Herbal medicines continue to be widely used as natural promoters of good health, as immune-modulators in recent years. This situation is directly related to the rapid growth of natural based products, the decrease of chemical synthesized products and as well as the increase of natural substance consumption. @*Objective@#The purpose of this survey was to study influence of Immunos herbal medicines on immune system in the experimental and preclinical circumstances.@*Materials and Methods@#The immune deficiency was to created by Azathioprine through 5 days in the white mice after that control group, preparation of Immunal, Salimon and Immunos 1, 2 were administrated appropriate doses by oral during 10 days. Then we collected blood and quantified number of white blood cells (K/µL), quantity of splenocyte (×106 cell/ml), amount of CD4+, CD8+ and IgM, IgA, Ig G (mg/ml) (Elisa Kit Assay: Catalog. No: WAM-568 (Elisa Reader, 450 nm)-WKEA MED SUPPLIES CORP) on the 5th, 10th days.@*Results@#All statistical analyses were conducted with SPSS version 20.0 software (IBM, Armonk, NY). One-way ANOVA was used to assess statistical significance between Immunos groups and days of observation. Mean values of white blood cells in blood, quantity of splenocyte, CD4+, CD8+ and IgM, IgG levels determined in the control and experimental groups. White blood cells level were significantly increased in the Immunos group compared with the control group by 55.6 percent (11.5±0.9 K/µL vs 5.1±0.51 K/µL, p<0.001) and number of splenocyte increased Immunos group compared with the control group by 60.6 % (352.2±23.5 ×106 cell/ml vs 138.6±23.5 ×106 cell/ml, p<0.01). Therefore, CD4+, CD8+ and IgM, IgG levels were significantly increased in the Immunos group compared with the control group by 0.71 to 8.8% (IgG: 11.47±0.42 vs 10.45±0.43 μg/ml, IgM: 11.33±0.81 vs 10.48±0.31 μg/ml, CD4+: 10.44±0635 vs 10.04±0.372 U/ml, CD8+: 9.75±1.02 vs 9.68±0.45 U/ml p<0.02).@*Conclusion@#It’s concluded that, Immunos preparation shows immune-stimulator effect in cellular immunity and humoral immunity in the case of immunosuppressant by Azathioprine.

Splenic lymphocyte immune response induced by intranasal immunization with recombinant Toxoplasma gondii profilin in mice / 中国血吸虫病防治杂志

Objective To observe the splenocytes immune response elicited by different concentrations of recombinant Toxo?plasma gondii profilin(rTgPRF)through the nasal route,and determine the optimal dose. Methods Fifty female BALB/c mice were randomly divided into 5 groups. The immunized groups were intranasally administered with 10,20,30μg or 40μg of rTgPRF that was separately dissolved in 20μl of phosphate?buffered saline(PBS)on days 0,14,and 21 respectively,while the control mice were given PBS solution instead. Two weeks after the last immunization,all mice were killed. Under asceptic conditions,the spleens from the immunized mice were dissected,and then the splenocyte proliferative responses in vitro were tested by CCK?8 kit. The levels of IFN?γ,IL?2,IL?4 and IL?10 of splenocyte culture supernatant were detected by ELISA. Re?sults Compared to the control group,the splenocytes from the 30μg and 40μg groups exhibited a significantly higher prolifer?ative response to rTgPRF(P<0.05),and SI from the 30μg rTgPRF group was higher than that from the 40μg group(P<0.05). The levels of IFN?γin all the immunized groups(P<0.05)and IL?2 in the 20,30μg and 40μg groups were significant?ly stronger than those in the control(P<0.05),and the 30μg group presented the highest concentrations of IFN?γ(P<0.01) and IL?2(P<0.01). There were no statistical differencesa mong the groups in the levels of IL?4 and IL?10. Conclusions The intranasal immunization with rTgPRF can induce the splenocyteproliferation and Th1?type mediated immunity. The best immu?nized dose is confirmed as 30μg.

Immune stimulatory activity of BRP-4, an acidic polysaccharide from an edible plant, Basella rubra L

Objective: To evaluated the immunomodulatory effect of BRP-4, an acidic polysaccharide from Basella rubra (. B. rubra) L on the macrophage activity. Methods: Phagocytic activity was determined by the ingestion of Latex Beads-Rabbit IgG-FITC using the fluorescent microscopy and flow cytometry analysis and nitric oxide production was measured using Griess reaction assay. Results: An enhanced production of NO was observed at 10 and 100 μg/mL of BRP-4. The phagocytic activity of macrophage was enhanced in BRP-4 treated RAW264.7 cells. BRP-4 combined with concanavalin A (Con A) provided obvious promotion and strengthening of the proliferation of the splenocytes. Conclusions: BRP-4, polysaccharide isolated from B. rubra, is suggested to activate macrophage function and stimulate splenocyte proliferation. The strong immunomodulatory activity of BRP-4 confirmed its good potential as an immunotherapeutic adjuvant.

Immune stimulatory activity of BRP-4, an acidic polysaccharide from an edible plant, Basella rubra L

OBJECTIVE@#To evaluated the immunomodulatory effect of BRP-4, an acidic polysaccharide from Basella rubra (B. rubra) L on the macrophage activity.@*METHODS@#Phagocytic activity was determined by the ingestion of Latex Beads-Rabbit IgG-FITC using the fluorescent microscopy and flow cytometry analysis and nitric oxide production was measured using Griess reaction assay.@*RESULTS@#An enhanced production of NO was observed at 10 and 100 μg/mL of BRP-4. The phagocytic activity of macrophage was enhanced in BRP-4 treated RAW264.7 cells. BRP-4 combined with concanavalin A (Con A) provided obvious promotion and strengthening of the proliferation of the splenocytes.@*CONCLUSIONS@#BRP-4, polysaccharide isolated from B. rubra, is suggested to activate macrophage function and stimulate splenocyte proliferation. The strong immunomodulatory activity of BRP-4 confirmed its good potential as an immunotherapeutic adjuvant.

Dynamic observation on splenocyte in mice immunized with recombinant Bi f idobacterium bi f idum (pGEX-S j26GST) of Schistosoma j aponicum / 中国人兽共患病学报

To observe the dynamic changes of splenocyte proliferation ,subsets and apoptosis in mice immunized with re-combinantBifidobacteriumbifidum(pGEX-Sj26GST)ofSchistosomajaponicum,themiceweresubcutaneously(SCgroup) and intranasally (IN group) immunized ,respectively .Four mice from each group were sacrificed in every 2 wk during 0-20 wk after immunization .Splenocyte proliferation was investigated by MTT colorimetric assay ,subsets of CD+4 and CD+8 T cells and apoptosis of splenocytes by FACsort flow cytometry .In SC group ,unstimulated and stimulated with S jAWA ,the level of splenocyte proliferation significantly increased at 4-20 wk after vaccination and increased markedly at 4-18 wk stimulated with ConA ,both of which peaked at 8 wk;in IN group ,the proliferation level of splenocyte cultured with SjAWA and ConA signif-icantly increased during the 4-18 wk ,2-10 wk and 14-18 wk ,2-8 wk and 12-18 wk ,respectively ,and all reached the maximum at the 4 wk after immunization (P＜0 .01 or P＜0 .05) .CD+4 subsets increased obviously during 2-14 wk ,2 wk and 6-16 wk re-spectively ,and reached the peak at 8 wk (P＜0 .01 or P＜0 .05) in both group ,while CD8+ subsets rose lightly during 2-20 wk in both group ,and reached the maximum at 8 wk (SC group) and 6 wk respectively (P>0 .05) .Whether unstimulated or stimulated with ConA ,the level of splenocyte apoptosis of which remarkably increased at 2-4 wk and 2-6 wk separately in SC group ,and both peaked at 2 wk (P＜0 .01 or P＜0 .05 );in IN group ,the level of splenocyte apoptosis all increased at 4 wk and reached the maximum at the same time .In summary ,by inducing the proliferation of splenocytes ,increasing CD4 + T cells and decreasing splenocyte apoptosis ,the rBb (pGEX-Sj26GST ) vaccine plays a critical role in the protective immune response .

Molecular Characterization and Mitogenic Activity of a Lectin from Purse Crab Philyra Pisum

A lectin from the hemolymph of purse crab, Philyra pisum, was found to have anti-proliferative activity on human lung cancer cells by our laboratory. In this study, P. pisum lectin (PPL) was molecularly characterized including molecular mass, amino acid sequences, amino acid composition, and the effects of metal ions, temperature, and pH on the activity. We found that PPL showed mitogenic activity on human lymphocytes and BALB/c mouse splenocytes. The mitogenic activity (maximum stimulation index, SI=9.57+/-0.59) of PPL on human lymphocytes was higher than that of a standard well-known plant mitogen, concanavalin A (maximum SI=8.80+/-0.59). The mitogenic activity mediated by PPL is required for optimum dosing, and higher or lower concentrations caused decreases in mitogenic response. PPL also induced mitogenic activity on mouse splenocytes, however, the maximum SI (1.77+/-0.09) on mouse splenocytes of PPL was lower than that (2.14+/-0.15) of concanavalin A. In conclusion, PPL is a metal ion-dependent monomer lectin with mitogenic activity, and could be used as a lymphocyte or splenocyte stimulator.

Effects of ionizing radiation on expression of P21 protein in Jurkat cell line and p21 gene in thymocytes and splenocytes of mice / 中华放射医学与防护杂志

Objective To investigate the effects of ionizing radiation on the expression of P21 protein in Jurkat cell line and p21 gene in thymocytes and splenocytes of mice.Methods Flow cytometry (FCM)was used to analyze the expression of P21 protein in Jurkat cells at 12 and 24 h after irradiation to 0,0.5,1.0,2.0,4.0,and 6.0 Gy.Real-time PCR was used to detect the expression of p21 gene in thymocytes and splenocytes of mice at4 and 24 h after irradiation to 0,0.5,1.0,2.0,4.0,and 6.0 Gy.Multi-staining was used to analyze the micronucleus rates of Rct in bone marrow.Results The expressions of P21 protein were increased in a dose-dependent manner during 0.5-4.0 Gy(t=-24.23--3.96,P<0.05),but decreased at 6.0 Gy at 12 and 24 h post-irradiation(t=-11.19,-14.50,P<0.05).The expressions of p2 1 gene in both thymocytes and splenocytes of mice were increased in dose-dependent manner in the range of 0-6.0 Gy(including 6.0 Gy)(t=-29.96-8.80,P<0.05),and reached to the peak at 6.0 Gy at 4 and 24 h post-irradiation(t=-11.84--3.42,P<0.05),except thymocytes at 4 h and 1.0 Gy post-irradiation(t=-3.42,P>0.05).Conclusions The expressions of P21 protein and p21 gene could be increased by X-ray irradiation.which shows good dosedependent manners in certain range of dose.

Immunoregulation of Murine Immunocytes to Bifidobacteria Strain Isolated from Feces of Healthy Korean Children: IL-10 Release and Proportional Change of CD4+CD25+ Cells

Bifidobacteria is one of the prototypes of probiotics bacteria, normally inhabitating the intestinal tract of humans. To search for a potent immunoregulatory Bifidobacteria strain, we screened the Bifidobacteria strains isolated from the feces of healthy Korean children. The mRNA or protein expression of an anti-inflammatory cytokine, IL-10, from mouse macrophages stimulated with live Bifidobacteria was examined. Of tested strains, Bifidobacteria A28 induced the highest IL-10 gene expression of murine macrophages. To probe immunoregulatory activity of the selected strain on the mice, we evaluated the proportional changes of CD4+CD25+ surface marker in the murine splenocytes. Flow cytometric analysis showed that the overall percentages of CD4+CD25+ cells in A28-treated splenocytes were higher than those of untreated splenocytes. In parallel, IL-10 release from A28-treated mouse peritoneal macrophages and splenocytes was significantly higher than that of untreated control cells. Collectively, the Bifidobacteria A28 strain isolated from the feces of healthy Korean children augments the mRNA or protein expression of IL-10 release from mouse peritoneal macrophages as well as the proportion of CD4+CD25+ cells of naive splenocytes. These provide in vitro scientific clues that Bifidobacteria A28 might be usable for anti-inflammatory disease such as inflammatory bowel disease (IBD).

Suppression of CD4+ T-Cells in the Spleen of Mice Infected with Toxoplasma gondii KI-1 Tachyzoites

Toxoplasma gondii KI-1, a recent new isolate from Korea, shows similar pathogenicity and infectivity to mice compared to the virulent RH strain. To understand characteristics of host immunity, including immune enhancement or suppression, we investigated proliferative responses and phenotypes of spleen cells. In addition, kinetics of IFN-gamma, a Th1 cytokine, was examined in BALB/c mice up to day 6 post-infection (PI). Intraperitoneal injection of mice with 103 KI-1 tachyzoites induced significant decreases (P < 0.05) in proliferative responses of spleen cells. This occurred at days 2-6 PI even when concanavalin A (con A) was added and when stimulated with KI-1 antigen, suggesting suppression of the immunity. CD4+ T-cells decreased markedly at day 2 PI (P < 0.05), whereas CD8+ T-cells, NK cells, and macrophages did not show significant changes, except a slight, but significant, increase of CD8+ T-cells at day 6 PI. The capacity of splenocytes to produce IFN-gamma by con A stimulation dropped significantly at days 2-6 PI. These results demonstrate that intraperitoneal injection of KI-1 tachyzoites can induce immunosuppression during the early stage of infection, as revealed by the decrease of CD4+ T-cells and IFN-gamma.

Changes of splenocyte subsets in mice immunized with recombinant Bb-Eg95-EgA31 vaccine against Echinococcus granulosus / 中国人兽共患病学报

To investigate the weight reduction of hydatid cyst and level of splenocyte subsets in mice immunized with recombinant Bifidobacteria bifidum (Bb) Bb-Eg95-EgA31 vaccine of Echinococcus granulosus (Eg) and challenged with Eg protoscoleces,BALB/c mice were immunized with the vaccine subcutaneously,intramuscularly,intranasally or orally.Then the mice were challenged with 50 Eg protoscoleces on 8~(th) week after vaccination and killed on 25~(th) week after infection.The weight of hydatid cyst was measured and the reduction of the weight was calculated.The spleens were collected for splenocytes preparation.CD_4~+ and CD_8~+ T cells were determined by flow cytometry(FCM),and the blank vector,Bb or MRS were set as control.The hydatid cyst weight was reduced by 45.33%,41.33%,70.67% and 62.67% respectively for the 4 immunization groups.Number of CD_4~+ and CD_8~+ subsets in the immunization groups was more than that in the control group.CD_4~+ and the ratio of CD_4~+/CD_8~+ in the intranasal or oral immunization group was much higher than that in the subcutaneous or intramuscular immunization group.It's concluded that CD_4~+ and CD_8~+T cells may play an important role in the protection induced by recombinant Eg95-EgA31 vaccine of Eg.In addition,intranasal and oral vaccinations could be the better ways for immunization.

Effects of Plant Water Extract Codonopsis Lanceolatae on Mouse Immune Cell Activation Ex Vivo

Codonopsis lanceolatae has been used as one of the traditional remedies as well as food source. However, few studies on their immunomodulating effects have been reported. We previously reported that ex vivo supplementation of Codonopsis lanceolatae water extracts enhanced splenocyte proliferation compared to the control group. In order to elucidate its ex vivo effect, six to seven week old balb/c mice were fed ad libitum on a chow diet and water extracts of Codonopsis lanceolatae were orally administrated every other day for four weeks at two different concentrations (50 and 500 mg/kg B.W.). After preparing the single cell suspension, the proliferation of splenocytes was determined by MTT (3-[4,5-dimethylthiazol-2-y]-2,5-diphenyl terazolium bromide) assay. The production of cytokine (IL-1beta, IL-6, TNF-alpha), secreted by macrophages stimulated with LPS or not, was detected by ELISA assay using a cytokine kit. After 48 hrs of incubation with the mitogen (ConA or LPS) stimulation, the mice splenocyte proliferation in experimental group was statistically increased at two different concentrations than that in control group. The cytokines production was more significantly enhanced at the lower supplementation (500 mg/kg B.W.) group rather than higher concentration (500 mg/kg B.W.) compared to the control group. The results of this study may suggest that the supplementation of water extract of plant mixture could regulate the immune function by increasing the splenocyte proliferation and enhance the immune function through regulating cytokine production capacity by activated macrophages in mice.

The Inhibitory Effect of Propofol on Splenocytes Proliferations to Lipopolysaccharide in BALB/c Mice: Based on the Measurement of BrdU Incorporation in vitro / 대한마취과학회지

BACKGROUND: Anesthetics have been suspected of impairing various aspects of the immune function either directly by affecting the function of immunocompetent cells or indirectly by modulating the stress response. Splenocytes play important roles in the cellular host defense against infection. In order to assess the immune modulatory effects of propofol, this study examined the cytotoxic and proliferative effects of propofol on splenocytes. METHODS: Splenocytes, as responders, were isolated from BALB/c mice (n = 10). The cells were pretreated with different propofol concentrations (0micrometer, 30micrometer, 100micrometer, 300micrometer) for 24 hours. The cytotoxic effect was assayed by the NADH dehydrogenase activity and the proliferation was evaluated by the level of 5-bromo-2'-deoxyunridine (BrdU) incorporation during DNA synthesis in the presence or absence of propofol, in addition to lipopolysaccharide (LPS, 1 microgram/ml) for mitogenic stimulation. A cell proliferation enzyme-linked immuno-sorbent assay (ELISA) system was used, and the stimulation index was calculated in the presence or absence of propofol. RESULTS: The percentage of the NADH dehydrogenase activity was changed by the propofol pretreatment (P < 0.001). LPS stimulation significantly decreased the NADH dehydrogenase activity at 100micrometer and 300micrometer compared with the propofol-added or pretreated cells (P < 0.05). The stimulation index to LPS was lower at concentrations of 100micrometer and 300micrometer than at 30micrometer, and proliferative response of splenocytes were completely abrogated by adding toxic concentrations (100micrometer) of propofol (P < 0.05). CONCLUSIONS: Neither cytotoxicity, as defined by the NADH dehydrogenase activity, nor a proliferative effect, as measured by the level of (BrdU) incorporation in the splenocytes, were affected by the clinical concentration of propofol.

Effects of Plant Water Extract Mixture Ixeris sonchifolia Hance, Oenanthevjavanica, Fagopyrum Esculentum Moench, Hizikia Fusiforme, Zingiber Officinale Roscoe on Mouse Immune Cell Activation Ex vivo

Ixeris sonchifolia Hance (Godulbaegi), Oenanthe javanica (Dolminari), Fagopyrum esculentum Moench (Buckwheat), Hizikia fusiforme (Seaweed Fusiforme) and Zingiber officinale Roscoe (Ginger) have been used respectively as one of folk remedies as well as food materials. However, reportedly few studies on their immunomodulating effects have been made, although it has been known from other preceding studies that the ex vivo supplementation of each Ish, Oj, Fem, Hf, Zor water extracts tends to enhance the proliferation of splenocyte in comparison to the control group. This study on the combined immunomodulative effect of water extract mixture of these five food materials (Ish + Oj + Fem + Hf + Zor) lasted covering seven or eight weeks. The old mice (balb/c) was fed ad libitum on chow diet, and the water extract of plant mixture was orally administrated every other day for four weeks at two different concentrations (50 and 500 mg/kg B.W). After preparing the single cell suspension, the proliferation of splenocyte was determined by MTT (3-[4,5-dimethylthiazol-2-y]-2,5- diphenyl terazolium bromide) assay. The production of cytokine (IL-1beta, IL-6, and TNF-alpha) which was secreted by macrophages stimulated with LPS or not was detected by ELISA assay using the cytokine kit. After the 48 hours of incubation with the mitogen (ConA or LPS) stimulation, the proliferation of the mice splenocyt in the experimental group statisticaly increased at both of two different concentrations in comparison to the control group. The cytokines production was more significantly enhanced at the lower supplementation (50 mg/kg B.W.) group than at the higher concentration (500 mg/kg B.W.). The result of this study may suggest that the supplementation of water extract of plant mixture can regulate and enhance the immune function by increasing the splenocyte proliferation and regulating the cytokine production capacity by the activated macrophages in mice.

Model establishment of Graves’disease by immunizing mice with hTSHR-activated splenocytes / 重庆医科大学学报

Objective:To establish an animal model of Graves’disease by immunizing syngeneic BALB/c mice with hTSHR-ac-tivated splenocytes. Methods:The female BALB/c mice(6～8 weeks) were randomly divided into 2 groups(group a and group b), which were immunized with recombinant plasmid pcDNA3.1/hTSHR and blank plasmid pcDNA3.1(+) respectively for 3 times at 3-week intervals.The animals were sacrificed at 18th week and their splenocytes were isolated.Other two groups of female syn- geneic BALB/c mice(6~8 weeks) were divided randomly into experimental and control groups, which received splenocytes from group a and group b respectively. Four weeks later, the thyroid glands were removed for histological examination and the sera were determined for Total T4,TSH and TRAb measurements. Results:The thyroid tissues under light microscope displayed dif- fuse hypercellularity with the follicular epithelia being tall cuboidal cells and reduced colloid in 44% of experimental mice with irregular apical poles protruding into the follicular space; there was minimal immune cell in the interstitium with hyper- aemia and edema. Electron microscope examination revealed that the follicular epithelia contained numerous mitochondria and rough endoplasmic reticula,and many long microvilli existed in the luminal surface.The serum level of TT4 in experimental mice was higher than that in control(7.13?1.02 vs 6.43?0.57 ?g/dL) (P

Effects of Deer Antler Extract on Serum IGF-I, Bone Growth and Splenocyte Proliferation in Growing Rats

Although it has traditionally known that deer antler and medicinal herbs extract contain some functional components for health promotion, the nutritional significance remains to be elucidated. This study examined the efficacy of deer antler extract (DA) , medicinal herbs extract (MH) and their mixture (DAMH) on serum IGF-I, bone growth with growing rats in vivo and splenocyte proliferation with spleen cells in vitro. Three week-old young female rats (Sprague-Dawley) were divided into 4 groups and then fed basal diet (AIN-93G) or experimental diets containing DA, MH, DAMH, respectively, for 7 weeks. We collected blood, liver, kidney, spleen, femur and tibia from rats. There was no significant difference in weight gain, but food intake increased in DA- and MH-fed groups. There were no signs of liver and kidney damage in the DA, MH and DAMH-fed groups compared to basal diet group. In femur and tibia, wet weights, breaking forces and bone minerals (Ca, Mg and Zn) were significantly higher in the DA-fed group than in the other groups. Serum alkaline phosphatase (ALP) , bone-specific alkaline phosphatase (BALP) activities were significantly lower in the DA, MH, DAMH-fed groups than in basal diet group. Also, serum insulin-like growth factor-I (IGF-I) concentrations were significantly increased in DA-fed group compared to the other groups. Therefore DA was shown to have an activity of bone growth promotion by increasing the IGF-I, a major bone growth factor. The deer antler extract showed an enhanced immune action on the primary cultured-cells from spleen of rats, representing that splenocytes were proliferated by lipopolysaccharide (LPS) , but not by concanavalin A (Con A) . These results indicate that deer antler extract has beneficial effects on bone growth via IGF-I and on splenocyte activation.