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Mulberry fruit prevents LPS-induced NF-κB/pERK/MAPK signals in macrophages and suppresses acute colitis and colorectal tumorigenesis in mice.

Qian Z, Wu Z, Huang L, Qiu H, Wang L, Li L, Yao L, Kang K, Qu J, Wu Y, Luo J, Liu JJ, Yang Y, Yang W, Gou D - Sci Rep (2015)

Bottom Line: In vitro, LPS-induced nitric oxide (NO) production was significantly inhibited by MBF extracts via suppressing the expression of proinflammatory molecules, including inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-1 beta (IL-β) and IL-6.In vivo, DSS-induced acute colitis was significantly ameliorated in MBF-fed mice as gauged by weight loss, colon morphology and histological damage.In addition, MBF-fed MUC2(-/-) mice displayed significant decrease in intestinal tumor and inflammation incidence compared to control diet-fed group.

View Article: PubMed Central - PubMed

Affiliation: Shenzhen Key Laboratory of Microbial Genetic Engineering, College of Life Sciences, Shenzhen University, Shenzhen, Guangdong, 518060, China.

ABSTRACT
Here, we investigated the impact of mulberry fruit (MBF) extracts on lipopolysaccharide (LPS)-induced inflammatory responses in RAW 264.7 macrophages, and the therapeutic efficacy of MBF diet in mice with dextran sulfate sodium (DSS)-induced acute colitis and MUC2(-/-) mice with colorectal cancer. In vitro, LPS-induced nitric oxide (NO) production was significantly inhibited by MBF extracts via suppressing the expression of proinflammatory molecules, including inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-1 beta (IL-β) and IL-6. Particularly, a dose-dependent inhibition on LPS-induced inflammatory responses was observed following treatment with MBF dichloromethane extract (MBF-DE), in which linoleic acid and ethyl linolenate were identified as two active compounds. Moreover, we elucidated that MBF-DE attenuated LPS-induced inflammatory responses by blocking activation of both NF-κB/p65 and pERK/MAPK pathways. In vivo, DSS-induced acute colitis was significantly ameliorated in MBF-fed mice as gauged by weight loss, colon morphology and histological damage. In addition, MBF-fed MUC2(-/-) mice displayed significant decrease in intestinal tumor and inflammation incidence compared to control diet-fed group. Overall, our results demonstrated that MBF suppressed the development of intestinal inflammation and tumorgenesis both in vitro and in vivo, and supports the potential of MBF as a therapeutic functional food for testing in human clinical trials.

No MeSH data available.


Related in: MedlinePlus

MBF-DE inhibits NF-κB p65 nuclear translocation in LPS-induced RAW 264.7 macrophage cells.Cells were pretreated with or without MBF-DE (200 μg/ml) for 12 h and then exposed to LPS (1 μg/ml) for 1 h. Then cells were fixed, permeabilized and processed using immunofluorescent staining for p65. Nuclei were stained with DAPI (a). The relative percentage of p65 translocation compared to LPS control was quantified based on three independent experiments (c). The protein level of p65 in nuclear and cytoplasmic fractions was determined by Western blotting (b) and the relative change was quantified (d). Results are shown as means ± SD of three independent experiments. **P < 0.01, ***P < 0.001 compared to LPS-induced control.
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f4: MBF-DE inhibits NF-κB p65 nuclear translocation in LPS-induced RAW 264.7 macrophage cells.Cells were pretreated with or without MBF-DE (200 μg/ml) for 12 h and then exposed to LPS (1 μg/ml) for 1 h. Then cells were fixed, permeabilized and processed using immunofluorescent staining for p65. Nuclei were stained with DAPI (a). The relative percentage of p65 translocation compared to LPS control was quantified based on three independent experiments (c). The protein level of p65 in nuclear and cytoplasmic fractions was determined by Western blotting (b) and the relative change was quantified (d). Results are shown as means ± SD of three independent experiments. **P < 0.01, ***P < 0.001 compared to LPS-induced control.

Mentions: In parallel, the nuclear translocation of NF-κB/p65 was measured using confocal microscopic analysis. As shown in Fig. 4, LPS stimulation in macrophages resulted in a dramatic increase in the translocation of p65 into the nucleus, which in turn was markedly suppressed after 12 h of MBF-DE pre-treatment. This observation was further confirmed by the Western blotting, i.e., LPS induced accumulation of p65 protein in nuclear was inhibited by MBF-DE treatment (Fig. 4b,d). Taken together, these data demonstrated that MBF-DE prevented not only NF-κB signals through blocking p65 nuclear translocation and IκBα phosphorylation but also MAPK/pERK activation via its phosphorylation.


Mulberry fruit prevents LPS-induced NF-κB/pERK/MAPK signals in macrophages and suppresses acute colitis and colorectal tumorigenesis in mice.

Qian Z, Wu Z, Huang L, Qiu H, Wang L, Li L, Yao L, Kang K, Qu J, Wu Y, Luo J, Liu JJ, Yang Y, Yang W, Gou D - Sci Rep (2015)

MBF-DE inhibits NF-κB p65 nuclear translocation in LPS-induced RAW 264.7 macrophage cells.Cells were pretreated with or without MBF-DE (200 μg/ml) for 12 h and then exposed to LPS (1 μg/ml) for 1 h. Then cells were fixed, permeabilized and processed using immunofluorescent staining for p65. Nuclei were stained with DAPI (a). The relative percentage of p65 translocation compared to LPS control was quantified based on three independent experiments (c). The protein level of p65 in nuclear and cytoplasmic fractions was determined by Western blotting (b) and the relative change was quantified (d). Results are shown as means ± SD of three independent experiments. **P < 0.01, ***P < 0.001 compared to LPS-induced control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4663626&req=5

f4: MBF-DE inhibits NF-κB p65 nuclear translocation in LPS-induced RAW 264.7 macrophage cells.Cells were pretreated with or without MBF-DE (200 μg/ml) for 12 h and then exposed to LPS (1 μg/ml) for 1 h. Then cells were fixed, permeabilized and processed using immunofluorescent staining for p65. Nuclei were stained with DAPI (a). The relative percentage of p65 translocation compared to LPS control was quantified based on three independent experiments (c). The protein level of p65 in nuclear and cytoplasmic fractions was determined by Western blotting (b) and the relative change was quantified (d). Results are shown as means ± SD of three independent experiments. **P < 0.01, ***P < 0.001 compared to LPS-induced control.
Mentions: In parallel, the nuclear translocation of NF-κB/p65 was measured using confocal microscopic analysis. As shown in Fig. 4, LPS stimulation in macrophages resulted in a dramatic increase in the translocation of p65 into the nucleus, which in turn was markedly suppressed after 12 h of MBF-DE pre-treatment. This observation was further confirmed by the Western blotting, i.e., LPS induced accumulation of p65 protein in nuclear was inhibited by MBF-DE treatment (Fig. 4b,d). Taken together, these data demonstrated that MBF-DE prevented not only NF-κB signals through blocking p65 nuclear translocation and IκBα phosphorylation but also MAPK/pERK activation via its phosphorylation.

Bottom Line: In vitro, LPS-induced nitric oxide (NO) production was significantly inhibited by MBF extracts via suppressing the expression of proinflammatory molecules, including inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-1 beta (IL-β) and IL-6.In vivo, DSS-induced acute colitis was significantly ameliorated in MBF-fed mice as gauged by weight loss, colon morphology and histological damage.In addition, MBF-fed MUC2(-/-) mice displayed significant decrease in intestinal tumor and inflammation incidence compared to control diet-fed group.

View Article: PubMed Central - PubMed

Affiliation: Shenzhen Key Laboratory of Microbial Genetic Engineering, College of Life Sciences, Shenzhen University, Shenzhen, Guangdong, 518060, China.

ABSTRACT
Here, we investigated the impact of mulberry fruit (MBF) extracts on lipopolysaccharide (LPS)-induced inflammatory responses in RAW 264.7 macrophages, and the therapeutic efficacy of MBF diet in mice with dextran sulfate sodium (DSS)-induced acute colitis and MUC2(-/-) mice with colorectal cancer. In vitro, LPS-induced nitric oxide (NO) production was significantly inhibited by MBF extracts via suppressing the expression of proinflammatory molecules, including inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-1 beta (IL-β) and IL-6. Particularly, a dose-dependent inhibition on LPS-induced inflammatory responses was observed following treatment with MBF dichloromethane extract (MBF-DE), in which linoleic acid and ethyl linolenate were identified as two active compounds. Moreover, we elucidated that MBF-DE attenuated LPS-induced inflammatory responses by blocking activation of both NF-κB/p65 and pERK/MAPK pathways. In vivo, DSS-induced acute colitis was significantly ameliorated in MBF-fed mice as gauged by weight loss, colon morphology and histological damage. In addition, MBF-fed MUC2(-/-) mice displayed significant decrease in intestinal tumor and inflammation incidence compared to control diet-fed group. Overall, our results demonstrated that MBF suppressed the development of intestinal inflammation and tumorgenesis both in vitro and in vivo, and supports the potential of MBF as a therapeutic functional food for testing in human clinical trials.

No MeSH data available.


Related in: MedlinePlus