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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 extracts inhibit LPS-induced up-regulation of proinflammatory regulators in RAW 264.7 macrophage cells.Cells were stimulated for 6 h with LPS (1 μg/mL) alone or together with MBF-DE, MBF-BE and MBF-EE at concentrations indicated. The mRNA levels of iNOS (a), COX-2 (b), IL-1β (c) and IL-6 (d) were analyzed by real-time qRT-PCR and the RPL14 was used to normalize data. The protein expression of iNOS and COX-2 (e,f) were determined by Western blotting and β-actin was used as an internal loading control. Results are shown as means ± SD of three independent experiments. * or #P < 0.05, ** or ##P < 0.01, ***P < 0.001 compared to LPS-induced control.
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f2: MBF extracts inhibit LPS-induced up-regulation of proinflammatory regulators in RAW 264.7 macrophage cells.Cells were stimulated for 6 h with LPS (1 μg/mL) alone or together with MBF-DE, MBF-BE and MBF-EE at concentrations indicated. The mRNA levels of iNOS (a), COX-2 (b), IL-1β (c) and IL-6 (d) were analyzed by real-time qRT-PCR and the RPL14 was used to normalize data. The protein expression of iNOS and COX-2 (e,f) were determined by Western blotting and β-actin was used as an internal loading control. Results are shown as means ± SD of three independent experiments. * or #P < 0.05, ** or ##P < 0.01, ***P < 0.001 compared to LPS-induced control.

Mentions: Proinflammatory mediators and cytokines such as iNOS, COX-2, IL-1β and IL-6 play important roles in regulating inflammatory response2728. To investigate whether or not the MBF-DE, MBF-EE and MBF-BE have anti-inflammatory activities in LPS stimulated macrophages, the expression of iNOS, COX-2, IL-1β and IL-6 was measured at both transcriptional and translational levels. Compared with the blank group, LPS induced up-regulation of mRNA levels of iNOS, COX-2, IL-1β and IL-6 were markedly inhibited by the treatment with MBF-DE, MBF-EE and MBF-BE at different concentrations (Fig. 2a–d). Similar inhibitory effects of MBF extracts on protein expression of these proinflammatory genes were also found in LPS stimulated macrophages (Fig. 2e,f and Supplementary Fig. 1). Among them, a significant dose-dependent manner was observed following the treatment with MBF-DE, i.e. the higher concentration of MBF-DE applied, the greater inhibitory effect on expression of these genes were shown (Fig. 2). These results illustrated that the expression of iNOS, COX-2, IL-1β and IL-6 in LPS-stimulated macrophages were suppressed by MBF extracts, in which the MBF-DE treatment displayed a concentration-dependent inhibitory effect. Therefore, MBF-DE was used for further study in following experiments hereafter, unless stated otherwise.


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 extracts inhibit LPS-induced up-regulation of proinflammatory regulators in RAW 264.7 macrophage cells.Cells were stimulated for 6 h with LPS (1 μg/mL) alone or together with MBF-DE, MBF-BE and MBF-EE at concentrations indicated. The mRNA levels of iNOS (a), COX-2 (b), IL-1β (c) and IL-6 (d) were analyzed by real-time qRT-PCR and the RPL14 was used to normalize data. The protein expression of iNOS and COX-2 (e,f) were determined by Western blotting and β-actin was used as an internal loading control. Results are shown as means ± SD of three independent experiments. * or #P < 0.05, ** or ##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

f2: MBF extracts inhibit LPS-induced up-regulation of proinflammatory regulators in RAW 264.7 macrophage cells.Cells were stimulated for 6 h with LPS (1 μg/mL) alone or together with MBF-DE, MBF-BE and MBF-EE at concentrations indicated. The mRNA levels of iNOS (a), COX-2 (b), IL-1β (c) and IL-6 (d) were analyzed by real-time qRT-PCR and the RPL14 was used to normalize data. The protein expression of iNOS and COX-2 (e,f) were determined by Western blotting and β-actin was used as an internal loading control. Results are shown as means ± SD of three independent experiments. * or #P < 0.05, ** or ##P < 0.01, ***P < 0.001 compared to LPS-induced control.
Mentions: Proinflammatory mediators and cytokines such as iNOS, COX-2, IL-1β and IL-6 play important roles in regulating inflammatory response2728. To investigate whether or not the MBF-DE, MBF-EE and MBF-BE have anti-inflammatory activities in LPS stimulated macrophages, the expression of iNOS, COX-2, IL-1β and IL-6 was measured at both transcriptional and translational levels. Compared with the blank group, LPS induced up-regulation of mRNA levels of iNOS, COX-2, IL-1β and IL-6 were markedly inhibited by the treatment with MBF-DE, MBF-EE and MBF-BE at different concentrations (Fig. 2a–d). Similar inhibitory effects of MBF extracts on protein expression of these proinflammatory genes were also found in LPS stimulated macrophages (Fig. 2e,f and Supplementary Fig. 1). Among them, a significant dose-dependent manner was observed following the treatment with MBF-DE, i.e. the higher concentration of MBF-DE applied, the greater inhibitory effect on expression of these genes were shown (Fig. 2). These results illustrated that the expression of iNOS, COX-2, IL-1β and IL-6 in LPS-stimulated macrophages were suppressed by MBF extracts, in which the MBF-DE treatment displayed a concentration-dependent inhibitory effect. Therefore, MBF-DE was used for further study in following experiments hereafter, unless stated otherwise.

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