Korean J Physiol Pharmacol.  2018 Sep;22(5):481-491. 10.4196/kjpp.2018.22.5.481.

Influence of rutin on the effects of neonatal cigarette smoke exposure-induced exacerbated MMP-9 expression, Th17 cytokines and NF-κB/iNOS-mediated inflammatory responses in asthmatic mice model

• Liu LL
• Zhang Y
• Zhang XF
• Li FH

• KMID: 2418998
• DOI: http://doi.org/10.4196/kjpp.2018.22.5.481

Abstract

Allergic asthma is one of the most enduring diseases of the airway. The T-helper cells and regulatory T-cells are critically involved in inflammatory responses, mucus hypersecretion, airway remodelling and in airway hyper-responsiveness. Cigarette smoke (CS) has been found to aggravate inflammatory responses in asthma. Though currently employed drugs are effective, associated side effects demand identification and development of novel drugs with negligible or no adverse effects. Rutin, plant-derived flavonoid has been found to possess antioxidant and anti-inflammatory effects. We investigated the ability of rutin to modulate T-cells and inhibit inflammation in experimentally-induced asthma in cigarette smoke exposed mice. Separate groups of neonatal mice were exposed to CS for 10 days from post-natal days 2 to 11. After 2 weeks, the mice were sensitized and challenged with ovalbumin (OVA). Treatment group were given rutin (37.5 or 75 mg/kg body weight) during OVA sensitization and challenge. Rutin treatment was found to significantly inhibit cellular infiltration in the airways and Th2 and Th17 cytokine levels as well. Flow cytometry revealed effectively raised CD4⁺CD25⁺Fox3⁺ Treg cells and supressed Th17 cell population on rutin treatment. Airway hyper-responsiveness observed following CS and OVA challenge were inhibited by rutin. NF-κB and iNOS, chief regulators of inflammatory responses robustly activated by CS and OVA were down-regulated by rutin. Rutin also inhibited the expression of matrix metalloproteinase 9, thereby aiding in prevention of airway remodelling in asthma thereby revealing to be a potent candidate in asthma therapy.

Keyword

Asthma; Cigarette smoke; Matrix metalloproteinase; Regulatory T cells; Rutin

MeSH Terms

Airway Remodeling
Animals
Asthma
Cytokines*
Flow Cytometry
Inflammation
Matrix Metalloproteinase 9
Mice*
Mucus
Ovalbumin
Ovum
Respiratory Hypersensitivity
Rutin*
Smoke*
T-Lymphocytes
T-Lymphocytes, Regulatory
Th17 Cells
Tobacco Products*
Cytokines
Matrix Metalloproteinase 9
Ovalbumin
Rutin
Smoke

Figure

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  Fig. 1 Rutin reduced airway resistance in lung function following CS and OVA challenge. Data are given as mean±SD where n=6. *Symbolizes p<0.05 related with control as determined by one way-ANOVA.

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  Fig. 2 Rutin improved lung compliance following CS and OVA challenge. Data are given as mean±SD where n=6. *Symbolizes p<0.05 related with control as determined by one way-ANOVA.

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  Fig. 3 Effect of Rutin on cell accumulation in BALF. Rutin reduced inflammatory cell infiltration in to BALF following CS and OVA challenge. Data are given as mean±SD where n=6. aDenotes statistical significance at p<0.05 related against control and b~gdenotes data within the same group that differ from each other at p<0.05 as calculated by one-way ANOVA followed by DMRT analysis.

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  Fig. 4 Rutin reduces OVA-specific IgE levels in serum and BALF. Rutin was found to significantly reduce the raised OVA-specific IgE in serum and in the BALF in mice exposed to CS and/or OVA. Data are given as mean±SD where n=6. aDenotes statistical significance at p<0.05 related against control and b~idenotes data within the same group that differ from each other at p<0.05 as calculated by one-way ANOVA followed by DMRT analysis.

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  Fig. 5 Rutin caused marked decrease in the levels of Th2 cytokines and increased the levels of IFN-γ. Data are given as mean±SD where n=6. aDenotes statistical significance at p<0.05 related against control and b~idenotes data within the same group that differ from each other at p<0.05 as calculated by one-way ANOVA followed by DMRT analysis.

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  Fig. 6 Rutin caused marked decrease in the levels of Th17 cytokines and increased the levels of IL-10. Data are given as mean±SD where n=6. adenotes statistical significance at p<0.05 related against control and b~idenotes data within the same group that differ from each other at p<0.05 as calculated by one-way ANOVA followed by DMRT analysis.

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  Fig. 7 Rutin regulates Th17 and CD4+CD25+Foxp3+ Treg cell populations. Data are given as mean±SD where n=6. aDenotes statistical significance at p<0.05 related against control and b~hdenotes data within the same group that differ from each other at p<0.05 as calculated by one-way ANOVA followed by DMRT analysis.

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  Fig. 8 Rutin was found to reduce inflammatory cell infiltration as in H&E staining (A) and reduce inflammation score (B). Data are given as mean±SD where n=6. aDenotes statistical significance at p<0.05 related against control and b~hdenotes data within the same group that differ from each other at p<0.05 as calculated by one-way ANOVA followed by DMRT analysis. (a) Control, (b) OVA+Rutin (75 mg), (c) CS+OVA+Rutin (75 mg), (d) CS+OVA+DEX.

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  Fig. 9 Rutin significantly reduce mucus hypersecretion and goblet cell hyperplasia as in PAFS staining (A and B). Data are given as mean±SD where n=6. aDenotes statistical significance at p<0.05 related against control and b~hdenotes data within the same group that differ from each other at p<0.05 as calculated by one-way ANOVA followed by DMRT analysis. (a) Control, (b) OVA, (c) OVA+Rutin (75 mg), (d) CS+OVA, (e) CS+OVA+Rutin (75 mg), (f) CS+OVA+DEX.

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  Fig. 10 Rutin modulates NF-κB/iNOS-mediated signaling and reduce MMP-9 expression. (A) L1, Control; L2, OVA; L3, OVA+Rutin (37.5 mg); L4, OVA+Rutin (75 mg); L5, CS+OVA; L6, CS+OVA+Rutin (37.5 mg); L7, CS+OVA+Rutin (75 mg); L8, CS+OVA+DEX. (B~D) Data are given as mean±SD where n=6. aDenotes statistical significance at p<0.05 related against control and b~hdenotes data within the same group that differ from each other at p<0.05 as calculated by one-way ANOVA followed by DMRT analysis.

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