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Dead Lactobacillus plantarum Stimulates and Skews Immune Responses toward T helper 1 and 17 Polarizations in RAW 264.7 Cells and Mouse Splenocytes

  • Lee, Hyun Ah;Kim, Hyunung;Lee, Kwang-Won;Park, Kun-Young
    • Journal of Microbiology and Biotechnology
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    • v.26 no.3
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    • pp.469-476
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    • 2016
  • 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 × 1011 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.

Ethanol extract of Callophyllis japonica enhances nitric oxide and tumor necrosis factor-alpha production in mouse macrophage cell line, RAW 264.7 cells

  • Ahn, Mee-Jung;Park, Dal-Soo;Yang, Won-Hyung;Go, Gyung-Min;Kim, Hyung-Min;Hyun, Jin-Won;Park, Jae-Woo;Shin, Taek-Yun
    • Advances in Traditional Medicine
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    • v.7 no.4
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    • pp.341-347
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    • 2007
  • Red seaweed (Callophyllis japonica) has long formed part of the diet of Asians, but the pharmacological properties of this plant have not been evaluated. In this study, we examined the effect of an ethanol extract of C. japonica on the generation of nitric oxide (NO) in RAW 264.7 cells. The C. japonica extract increased the generation of NO and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), which were detected by the Griess method and an enzyme-linked immunosorbent assay, respectively. The increased production of NO by C. japonica extract was inhibited by $N^G$-monomethyl-L-arginine ($100{\mu}M$), a specific inhibitor of NO production in the L-arginine-dependent pathway, and by the nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) inhibitor, pyrrolidine dithiocarbamate ($10-100{\mu}M$) in a dose-dependent manner. These findings demonstrate that C. japonica extract stimulates the production of NO and $TNF-{\alpha}$ in RAW 264.7 cells through the activation of $NF-{\kappa}B$ and that this extract might also inhibit the growth of the human leukemic cells.

Effects of Eucommiae Cortex on Osteoblast-like Cell Proliferation and Osteoclast Inhibition

  • Ha, Hyek-Yung;Ho, Jinn-Yung;Shin, Sun-Mi;Kim, Hye-Jin;Koo, Sung-Ja;Kim, In-Ho;Kim, Chung-Sook
    • Archives of Pharmacal Research
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    • v.26 no.11
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    • pp.929-936
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    • 2003
  • Methanol extract (MeOH), n-hexane (Hx), chloroform ($CHCl_3$), ethyl acetate (EA), butanol (BuOH) and aqueous ($H_2O$) fractions of Eucommiae Cortex including geniposidic acid (GA), geniposide (GP) and aucubin (AU) were tested for their therapeutic efficacy on osteoporosis. The contents of GA, GP and AU in the cortex and leaf of Eucommia ulmoides Oliver were quantified by HPLC. The effect of Eucommiae Cortex on the induction of growth hormone (GH) release was studied by using rat pituitary cells. The proliferation of osteoblast-like cells increased by herbal extracts was assayed using a tetrazolium (MTT), alkaline phosphatase (ALP) activity, and [$^3H$]-proline incorporation assays. The inhibition of osteoclast was studied by using the coculture of mouse bone marrow cells and ST-2 cells. As a result, the GA, GP and AU were present in the cortex more than in the leaf of E. ulmoides Oliver. The MeOH (1mg/mL), Hx, $CHCl_3$ and EA fractions (each 20 $\mu$ g/mL) had potent induction of GH release. The $CHCl_3$ exhibited the potent proliferation of osteoblasts. The AU, GP and GA were increased proliferation of osteoblasts. In addition, GA ($IC_{50}: 4.43{\times}10^{-7}$M), AU and GP were significantly inhibited proliferation of osteoclast. In summary, it is thought that the components in a part of the fractions of Eucommiae Cortex participate in each step of mechanism for activating osteoblast to facilitate osteogenesis, and suppress osteoclast activity to inhibit osteolysis.

Phospholipid polymer can reduce cytotoxicity of poly (lactic acid) nanoparticles in a high-content screening assay

  • Kim, Hyung Il;Ishihara, Kazuhiko
    • Biomaterials and Biomechanics in Bioengineering
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    • v.1 no.2
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    • pp.95-104
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    • 2014
  • The objective of this study was to evaluate the cytotoxicity of poly (lactic acid) (PLA) nanoparticles. We used a water-soluble, amphiphilic phospholipid polymer, poly (2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate) (PMB30W), as a stabilizer for the PLA nanoparticles. The PLA nanoparticles and PMB30W-modified PLA (PLA/PMB30W) nanoparticles were prepared by evaporating tetrahydrofuran (THF) from its aqueous solution. Precipitation of the polymers from the aqueous solution produced PLA and PLA/PMB30W nanoparticles with a size distribution of $0.4-0.5{\mu}m$. The partial coverage of PMB30W on the surface of the PLA/PMB30W nanoparticles was confirmed by X-ray photoelectron spectroscopy (XPS) and dynamic light-scattering (DLS). A high-content automated screening assay (240 random fields per group) revealed that the PLA nanoparticles induced apoptosis in a mouse macrophage-like cell line (apoptotic population: 73.9% in 0.8 mg PLA/mL), while the PLA/PMB30W nanoparticles remained relatively non-hazardous in vitro (apoptotic population: 13.8% in 0.8 mg PLA/mL). The reduction of the apoptotic population was attributed to the phosphorylcholine groups in the PMB30W bound to the surface of the nanoparticle. In conclusion, precipitation of PLA in THF aqueous solution enabled the preparation of PLA nanoparticles with similar shapes and size distribution but different surface characteristics. PMB30W was an effective stabilizer and surface modifier, which reduced the cytotoxicity of PLA nanoparticles by enabling their avoidance of the mononuclear phagocyte system.

Antioxidative Activity of Streptomyces sp. BH-405 on Macrophage Mediated Modification of Human Low Density Lipoprotein[LDL) (Macrophage 유도 사람 Low Density Lipoprotein[LDL]의 수식에 대한 Streptomyces sp. BH-405 배양액의 항산화 활성)

  • 류병호;박종옥;김희숙;김민정
    • KSBB Journal
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    • v.15 no.2
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    • pp.156-161
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    • 2000
  • This study was designed to investigate the antioxidative activity on oxidation of human low density lipoprotein(LDL) of band 2 fractionated from culture broth of Streptomyces sp. BH-405. Antioxidative activity of band 2 obtained from fractionation of BH-405 culture purification was measured against $Cu^{2+}$ mediated human LDL oxidation by thiobarbituric acid reactive substance. $CuSO_4$ mediated oxidation of LDL was degraded at a much higher rate than native LDL. Band 2 at a concentration of 100 or 200 !lg/mL inhibited the oxidation of LDL induced by $CuSO_4$, The formation of conjugated dienes induced in the presence of 5 !1M CuS04 of the mouse macrophage and J744. The electrophoretic mobility of the LDL in addition of $200\mu\textrm{g}$ band 2 in the presence of $5\mu\textrm{m}$ $CuSO_4$ was lower than that of native LDL. LDL modified by copper mediated or cell mediated uptake was degraded by macrophage at much greater than native LDL, and band 2 was found as potential inhibitor of modification of 125I-labelled LDL by macrophage. phage.

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Effects of Lubiprostone on Pacemaker Activity of Interstitial Cells of Cajal from the Mouse Colon

  • Jiao, Han-Yi;Kim, Dong Hyun;Ki, Jung Suk;Ryu, Kwon Ho;Choi, Seok;Jun, Jae Yeoul
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.4
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    • pp.341-346
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    • 2014
  • Lubiprostone is a chloride ($Cl^-$) channel activator derived from prostaglandin $E_1$ and used for managing constipation. In addition, lubiprostone affects the activity of gastrointestinal smooth muscles. Interstitial cells of Cajal (ICCs) are pacemaker cells that generate slow-wave activity in smooth muscles. We studied the effects of lubiprostone on the pacemaker potentials of colonic ICCs. We used the whole-cell patch-clamp technique to determine the pacemaker activity in cultured colonic ICCs obtained from mice. Lubiprostone hyperpolarized the membrane and inhibited the generation of pacemaker potentials. Prostanoid $EP_1$, $EP_2$, $EP_3$, and $EP_4$ antagonists (SC-19220, PF-04418948, 6-methoxypyridine-2-boronc acid N-phenyldiethanolamine ester, and GW627368, respectively) did not block the response to lubiprostone. L-NG-nitroarginine methyl ester (L-NAME, an inhibitor of nitric oxide synthase) and 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ, an inhibitor of guanylate cyclase) did not block the response to lubiprostone. In addition, tetraethylammonium (TEA, a voltage-dependent potassium [$K^+$] channel blocker) and apamin (a calcium [$Ca^{2+}$]-dependent $K^+$ channel blocker) did not block the response to lubiprostone. However, glibenclamide (an ATP-sensitive $K^+$ channel blocker) blocked the response to lubiprostone. Similar to lubiprostone, pinacidil (an opener of ATP-sensitive $K^+$ channel) hyperpolarized the membrane and inhibited the generation of pacemaker potentials, and these effects were inhibited by glibenclamide. These results suggest that lubiprostone can modulate the pacemaker potentials of colonic ICCs via activation of ATP-sensitive $K^+$ channel through a prostanoid EP receptor-independent mechanism.

Anti-adipogenesis Effects of 3 Herbal Formula on Blood Stasis (어혈처방 3종이 지방전구세포 분화에 미치는 영향)

  • Lee, Hoyoung;Shim, Eun Hyoung;Lee, Myeong Soo;Lee, Ju Ah
    • The Korea Journal of Herbology
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    • v.31 no.6
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    • pp.29-35
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    • 2016
  • Objectives : Blood stasis (BS) is related to be caused by blood circulation and stagnation which are cancer, atherosclerosis and hyperlipidemia in traditional medicine. We extracted 3 kinds of BS formula; Seogakjihwang-tang (SGT), Tonggyuhawlhyul-tang (THT), Hyulbuchukeo-tang (HCT). This study was conducted to investigate whether the 3 kinds of herbal formula extracts have inhibitory efficacy association with anti-adipogenesis. Methods : To investigate the anti-adipogenesis, we used the mouse fibroblast cell line, 3T3-L1 which differentiated into adipocytes in response to insulin, IBMX and dexamethasone (MDI). Cytotoxicity of herbal formula extracts were examined by CCK-8 kit. Intracellular lipid droplets were detected by Oil-Red-O staining. Triglyceride (TG) and leptin were measure using elisa kit. Results : The yield of water extracts was 14.62% (SGT), 21.27% (THT), 20.02% (HCT). Lipid accumulation was reduced significantly by 3 kinds of herbal formula compared to control. Especially, THT and HCT decreased lipid droplet, respectively at all concentration. The TG and leptin were also inhibited by 3 kinds of herbal formula. The IC50 of TG were $280.51{\mu}g/m{\ell}$ (SGT), $52.62{\mu}g/m{\ell}$ (THT), $313.99{\mu}g/m{\ell}$ (HCT). The IC50 of leptin were $348.76{\mu}g/m{\ell}$ (SGT), $164.02{\mu}g/m{\ell}$ (THT), $257.00{\mu}g/m{\ell}$ (HCT). THT was better than other herbal formula on anti-adipogenesis. Conclusion : kinds of herbal formula inhibited adipogenesis in MDI-induced 3T3-L1 adipocytes, as indicated by the significant reduction in TG and leptin concentration without cytotoxicity. Therefore, 3 kinds of herbal formula for BS might act as a therapeutic agent for preventing lipid diseases, such as obesity and atherosclerosis.

Evaluation of physical property and cytotoxicity of resin infiltrant based on a triethylene glycol dimethacrylate (TEGDMA)

  • Min, Ji-Hyun;Roh, Ji-Yeon;Kim, Ki-Rim
    • Journal of Korean society of Dental Hygiene
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    • v.19 no.2
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    • pp.173-181
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    • 2019
  • Objectives: The resin infiltration technique is a promising alternative therapy for arresting the early dental caries. However, there are very few reports on the safety and biocompatibility of this technique. We evaluated various properties of resin infiltrant (RI) based on a triethylene glycol dimethacrylate (TEGDMA).The water sorption (Wsp) and water solubility (Wsl) was assessed. Additionally, the cytotoxicity of RI against both animal and human fibroblast cell lines was investigated. Methods: The RI of the $Icon^{(R)}$, the first product developed for resin infiltration, is mainly composed of TEGDMA in the resin matrix. The Wsp and Wsl for the RI were measured in accordance with ISO 4049 specifications. Fourier-transform infrared spectroscopy (FTIR) was used for analyzing the polymerization before and after curing of RI. The cytotoxicity of RI against the mouse fibroblasts (L929) and human gingival fibroblasts (hTERT-hNOF) was evaluated using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and the data were analyzed using one-way analysis of variance. Results: Wsp and Wsl of the RI specimens were $53.37{\mu}g/mm^3$ and $10.6{\mu}g/mm^3$, respectively. FTIR analysis revealed a slightly higher degree of curing with longer irradiation time. The degree of conversion for RI was high (80.9%) after 40 seconds of light curing. There was a significant decrease in the viability of L929 and hTERT-hNOF cells at RI extraction solution concentrations above 50%, respectively, compared to that in the negative control (p< 0.05). Conclusions: Even though the RI exhibited positive effect on the early prevention of dental caries, the clinicians should also consider the toxicity of RI on periodontal tissues.

A study on Anti-diabetic Mechanism of Ethanol Extract of Dendrobii Herba (석곡 에탄올 추출물의 항당뇨 약리기전에 관한 연구)

  • Park, Myung-ji;Lee, Yeoung-Ju
    • Journal of Digital Convergence
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    • v.17 no.7
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    • pp.275-284
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    • 2019
  • Antidolary active and anti-sugar mechanisms of the ova family (石斛; Dendrobii herba) ethanol extract (EED) were investigated. The EED was administered orally four times a day in a diabetic mouse induced by strepto Joe Toshin to reveal and reveal its pharmacological miracle through experimental studies that reduce the liver function of empty blood sugar, glythamic oxal acetate levels, insulin levels and glutamic acid trans aminaase and glutamic acid pyruvic acid trans amine. EED increased insulin secretion by glucose in RINm5F beta cells as well as intraperitoneal glucose intakes in L6 muscle cells. Thus, EED has shown great promise in displaying anti-diabetes activity not only by increasing insulin secretion but also by increasing intakes per cell, and hopes that future research on pharmacological mechanisms for quartz (Dendrobii herba) ethanol extract will be more active and contribute greatly to the treatment of diabetes.

Antioxidative and Antitumor Activities of Crude Polysaccharide Fraction from P leurotus eryngii (큰느타리버섯(Pieurotus eryngii) 조다당체 분획의 항산화 및 항종양활성)

  • Kim, Jae-Yong;Kang, Hye-In;Park, Kyung-Uk;Moon, Kwang-Deog;Lee, Sang-Dae;Cho, Sook-Hyun;Wee, Jae-Joon;Kyung, Jong-Soo;Song, Yong-Beom;Seo, Kwon-Il
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.10
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    • pp.1589-1593
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    • 2004
  • The aims of this study were to investigate the antioxidative and antitumor effects of crude polysaccharide fraction from Pleurtus eryngii (CPPE). CPPE inhibited autoxidation of linoleic acid at 1,000 $\mu$g/mL concentration, and the inhibitory rates of lipid peroxidation in rat liver microsome were 5.28, 9.40, and 32.5% at 10, 100, and 1,000 $\mu$g/mL concentrations, respectively. After treatment with CPPE for 72 hours, the inhibitory rates against MCF-7, A549 and AGS cell lines showed 42.3, 33.4 and 26.7% at concentration of 1,000 $\mu$g/mL, respectively. Results of CPPE treatment at 100 and 300 mg/kg/day for 7 days in sarcoma-180 bearing-mice showed survival rates of 70 and 90%, respectively. Body weights of mice treated with CPPE were significantly decreased when compared with the control.