Korean J Physiol Pharmacol.  2015 Mar;19(2):183-189. 10.4196/kjpp.2015.19.2.183.

Foeniculum vulgare Mill. Protects against Lipopolysaccharide-induced Acute Lung Injury in Mice through ERK-dependent NF-kB Activation

• Lee HS
• Kang P
• Kim KY
• Seol GH

• KMID: 2285571
• DOI: http://doi.org/10.4196/kjpp.2015.19.2.183

Abstract

Foeniculum vulgare Mill. (fennel) is used to flavor food, in cosmetics, as an antioxidant, and to treat microbial, diabetic and common inflammation. No study to date, however, has assessed the anti-inflammatory effects of fennel in experimental models of inflammation. The aims of this study were to investigate the anti-inflammatory effects of fennel in model of lipopolysaccharide (LPS)-induced acute lung injury. Mice were randomly assigned to seven groups (n=7~10). In five groups, the mice were intraperitoneally injected with 1% Tween 80-saline (vehicle), fennel (125, 250, 500micro l/kg), or dexamethasone (1 mg/kg), followed 1 h later by intratracheal instillation of LPS (1.5 mg/kg). In two groups, the mice were intraperitoneally injected with vehicle or fennel (250microl/kg), followed 1 h later by intratracheal instillation of sterile saline. Mice were sacrificed 4 h later, and bronchoalveolar lavage fluid (BALF) and lung tissues were obtained. Fennel significantly and dose-dependently reduced LDH activity and immune cell numbers in LPS treated mice. In addition fennel effectively suppressed the LPS-induced increases in the production of the inflammatory cytokines interleukin-6 and tumor necrosis factor-alpha, with 500microl/kg fennel showing maximal reduction. Fennel also significantly and dose-dependently reduced the activity of the proinflammatory mediator matrix metalloproteinase 9 and the immune modulator nitric oxide (NO). Assessments of the involvement of the MAPK signaling pathway showed that fennel significantly decreased the LPS-induced phosphorylation of ERK. Fennel effectively blocked the inflammatory processes induced by LPS, by regulating pro-inflammatory cytokine production, transcription factors, and NO.

Keyword

ERK; Foeniculum vulgare Mill.; LPS; TNF-alpha

MeSH Terms

Acute Lung Injury*
Animals
Bronchoalveolar Lavage Fluid
Cell Count
Cytokines
Dexamethasone
Foeniculum*
Inflammation
Interleukin-6
Lung
Matrix Metalloproteinase 9
Mice*
Models, Theoretical
NF-kappa B*
Nitric Oxide
Phosphorylation
Transcription Factors
Tumor Necrosis Factor-alpha
Cytokines
Dexamethasone
Interleukin-6
Matrix Metalloproteinase 9
NF-kappa B
Nitric Oxide
Transcription Factors
Tumor Necrosis Factor-alpha

Figure

• 

  Fig. 1 Effect of fennel on lactate dehydrogenase (LDH) activity in BALF of LPS-treated mice. Mice were intratracheally administered LPS (1.5 mg/kg) 1 h after intraperitoneal injection of 1% Tween 80-saline (vehicle), fennel (125, 250, 500µl/kg), or DEX (1 mg/kg). The activity of LDH in BALF was measured to evaluate cell damage. Data are reported as mean±S.E.M. (n=7~10 per group). ###p<0.001 compared with vehicle group; **p<0.01, ***p<0.001 compared with the vehicle+LPS group.

• 

  Fig. 2 Effects of fennel on cell numbers in BALF of LPS-treated mice. The numbers of total cells, neutrophils, macrophages, and lymphocytes in BALF were analyzed. Data are reported as mean±S.E.M. (n=7~10 per group). #p<0.05, ##p<0.01, ###p<0.001 compared with the vehicle group; *p<0.05, **p<0.01, ***p<0.001 compared with the vehicle+LPS group.

• 

  Fig. 3 Effect of fennel on the histopathology of lung tissues in LPS-treated mice. Fennel (500µl/kg) or DEX (1 mg/kg) was administered intraperitoneally to mice 1 h prior to LPS treatment. Lung sections from each group were stained with hematoxylin and eosin (H&E) (×200). (A) Vehicle group, (B) Vehicle+LPS group, (C) Fennel+LPS group, (D) DEX+ LPS group.

• 

  Fig. 4 Effects of fennel on (A) IL-6 and (B) TNF-α expression in the BALF of LPS-treated mice. IL-6 and TNF-α in BALF were analyzed by ELISA. Data are reported as mean±S.E.M. (n=7~10 per group). ##p<0.01, ###p<0.001 compared with the vehicle group; *p<0.05, ***p<0.001 compared with the vehicle+LPS group.

• 

  Fig. 5 Effect of fennel on MMP-9 activity in LPS-treated mice. Relative MMP-9 activity in BALF was analyzed by zymography followed by scanning densitometry. Data are reported as mean±S.E.M. (n=7~10 per group). ##p<0.01, ###p<0.001 compared with the vehicle group; *p<0.05, **p<0.01, ***p<0.001 compared with the vehicle+LPS group.

• 

  Fig. 6 Effect of fennel on NO production in the BALF of LPS-treated mice. NO concentrations in BALF were measured by nitrite assays. Data are reported as mean±S.E.M. (n=7~10 per group). ###p<0.001 compared with the vehicle group; **p<0.01, ***p<0.001 compared with the vehicle+LPS group.

• 

  Fig. 7 Effect of fennel on NF-κB activation in LPS-treated mice. Nuclear and cytosolic extracts in lung tissue were fractionated and the expression of NF-κB p65 (A) and IκB-α (B) proteins in nuclear and cytosolic extracts, respectively, were assessed by western blotting. Lamin B and GAPDH were used as internal controls. Data are reported as mean±S.E.M. (n=7~10 per group). ##p<0.01, ###p<0.001 compared with the vehicle group; *p<0.05, ***p<0.001 compared with the vehicle+LPS group.

• 

  Fig. 8 Effect of fennel on the MAPK signaling pathway in LPS-treated mice. Lung tissues were analyzed by western blotting with antibodies to p-ERK (A), p-p38 (C), and p-JNK (E), and quantitative protein expression was normalized to ERK (B), p38 (D), and JNK (F), respectively. Data are reported as mean±S.E.M. (n=7~10 per group). #p<0.05, ###p<0.001 compared with the vehicle group; *p<0.05, **p<0.01 compared with the vehicle+LPS group.

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