Tuberc Respir Dis.  2012 Oct;73(4):204-209.

Effect of Chrysin on Gene Expression and Production of MUC5AC Mucin from Cultured Airway Epithelial Cells

Affiliations
  • 1Department of Pharmacology, Chungnam National University School of Medicine, Daejeon, Korea. LCJ123@cnu.ac.kr
  • 2Department of Orthopedic Surgery, Chungnam National University Hospital, Chungnam National University School of Medicine, Daejeon, Korea.
  • 3Pulmonology Section, Department of Internal Medicine, Chungnam National University Hospital, Chungnam National University School of Medicine, Daejeon, Korea.

Abstract

BACKGROUND
We investigated whether chrysin affected MUC5AC mucin production and gene expression induced by phorbol ester (phorbol 12-myristate 13-acetate, PMA) or epidermal growth factor (EGF) from human airway epithelial cells.
METHODS
Confluent NCI-H292 cells were pretreated with varying concentrations of chrysin for 30 minutes, and were then stimulated with PMA and EGF for 24 hours, respectively. MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay.
RESULTS
Concentrations of 10microM and 100microM chrysin were found to inhibit the production of MUC5AC mucin protein induced by PMA; A concentration of 100microM chrysin also inhibited the production of MUC5AC mucin protein induced by EGF; 100microM chrysin inhibited the expression of MUC5AC mucin gene induced by PMA or EGF. The cytotoxicity of chrysin was checked by lactate dehydrogenase assay, and there was no cytotoxic effect observed for chrysin.
CONCLUSION
These results suggest that chrysin can inhibit mucin gene expression and the production of mucin protein by directly acting on airway epithelial cells.

Keyword

Epithelial Cells; Mucins; Chrysin

MeSH Terms

Epidermal Growth Factor
Epithelial Cells
Flavonoids
Gene Expression
Humans
L-Lactate Dehydrogenase
Mucins
Phorbols
Polymerase Chain Reaction
Reverse Transcription
Epidermal Growth Factor
Flavonoids
L-Lactate Dehydrogenase
Mucins
Phorbols

Figure

  • Figure 1 Effect of chrysin on phorbol 12-myristate 13-acetate (PMA)-induced MUC5AC mucin production from NCI-H292 cells were then stimulated with PMA (10 ng/mL) for 24 hours. Cell lysates were collected for measurement of MUC5AC mucin production by enzyme-linked immunosorbent assa. Means of individual groups were converted to percentages of the control and were expressed as mean±SEM. The difference between groups was assessed using one-way ANOVA and Duncan's multiple range test. p<0.05 was considered as statistically significant. Each bar represents a mean±SEM of 3 culture wells, as compared to the control, set at 100%. *Significantly different from control (p<0.05). †Significantly different from PMA alone (p<0.05). PMA: phorbol 12-myristate 13-acetate; Cont: control. Concentration unit is µM.

  • Figure 2 Effect of chrysin on EGF-induced MUC5AC mucin production from NCI-H292 cells. NCI-H292 cells were pretreated with varying concentrations of chrysin for 30 minutes and were then stimulated with EGF (25 ng/mL) for 24 hours. Cell lysates were collected for measurement of MUC5AC mucin production by enzyme-linked immunosorbent assay. Means of individual group were converted to percentages of the control and were expressed as mean±SEM. The difference between groups was assessed using one-way ANOVA and Duncan's multiple range test. p<0.05 was considered as statistically significant. Each bar represents a mean±SEM of 3 culture wells, as compared to the control, set at 100%. *Significantly different from control (p<0.05). †Significantly different from EGF alone (p<0.05). EGF: epidermal growth factor; Cont: control. Concentration unit is µM.

  • Figure 3 Effect of chrysin on PMA-induced MUC5AC gene expression from NCI-H292 cells. NCI-H292 cells were pretreated with varying concentrations of chrysin for 30 minutes and were then stimulated with PMA (10 ng/mL) for 24 hours. MUC5AC gene expression was measured by RT-PCR. Rig/S15 rRNA, which encodes a small ribosomal subunit protein, a housekeeping gene that was constitutively expressed, was used as a quantitative control. PMA: phorbol 12-myristate 13-acetate; RT-PCR: reverse transcription polymerase chain reaction; Cont: control; Chr: chrysin. Concentration unit is µM.

  • Figure 4 Effect of chrysin on EGF-induced MUC5AC gene expression from NCI-H292 cells. NCI-H292 cells were pretreated with varying concentrations of chrysin for 30 min and were then stimulated with EGF (25 ng/mL) for 24 hours. MUC5AC gene expression was measured by RT-PCR. Rig/S15 rRNA, which encodes a small ribosomal subunit protein, a housekeeping gene that was constitutively expressed, was used as a quantitative control. EGF: epidermal growth factor; RT-PCR: reverse transcription polymerase chain reaction; Cont: control; Chr: chrysin. Concentration unit is µM.


Reference

1. Voynow JA, Rubin BK. Mucins, mucus, and sputum. Chest. 2009. 135:505–512.
2. Yuan-Chen Wu D, Wu R, Reddy SP, Lee YC, Chang MM. Distinctive epidermal growth factor receptor/extracellular regulated kinase-independent and -dependent signaling pathways in the induction of airway mucin 5B and mucin 5AC expression by phorbol 12-myristate 13-acetate. Am J Pathol. 2007. 170:20–32.
3. Rogers DF, Barnes PJ. Treatment of airway mucus hypersecretion. Ann Med. 2006. 38:116–125.
4. Lee CJ, Lee JH, Seok JH, Hur GM, Park YC, Seol IC, et al. Effects of baicalein, berberine, curcumin and hesperidin on mucin release from airway goblet cells. Planta Med. 2003. 69:523–526.
5. Heo HJ, Lee SY, Lee MN, Lee HJ, Seok JH, Lee CJ. Genistein and curcumin suppress epidermal growth factor-induced MUC5AC mucin production and gene expression from human airway epithelial cells. Phytother Res. 2009. 23:1458–1461.
6. Lee HJ, Lee SY, Lee MN, Kim JH, Chang GT, Seok JH, et al. Inhibition of secretion, production and gene expression of mucin from cultured airway epithelial cells by prunetin. Phytother Res. 2011. 25:1196–1200.
7. Jang IM. Treatise on Asian herbal medicines. 2003. Seoul: Haksul-pyunsu-kwan in Research Institute of Natural Products of Seoul National University.
8. Du Q, Gu X, Cai J, Huang M, Su M. Chrysin attenuates allergic airway inflammation by modulating the transcription factors T-bet and GATA-3 in mice. Mol Med Report. 2012. 6:100–104.
9. Shao JJ, Zhang AP, Qin W, Zheng L, Zhu YF, Chen X. AMP-activated protein kinase (AMPK) activation is involved in chrysin-induced growth inhibition and apoptosis in cultured A549 lung cancer cells. Biochem Biophys Res Commun. 2012. 423:448–453.
10. Sultana S, Verma K, Khan R. Nephroprotective efficacy of chrysin against cisplatin-induced toxicity via attenuation of oxidative stress. J Pharm Pharmacol. 2012. 64:872–881.
11. Li JD, Dohrman AF, Gallup M, Miyata S, Gum JR, Kim YS, et al. Transcriptional activation of mucin by Pseudomonas aeruginosa lipopolysaccharide in the pathogenesis of cystic fibrosis lung disease. Proc Natl Acad Sci U S A. 1997. 94:967–972.
12. Takeyama K, Dabbagh K, Lee HM, Agusti C, Lausier JA, Ueki IF, et al. Epidermal growth factor system regulates mucin production in airways. Proc Natl Acad Sci U S A. 1999. 96:3081–3086.
13. Shao MX, Ueki IF, Nadel JA. Tumor necrosis factor alpha-converting enzyme mediates MUC5AC mucin expression in cultured human airway epithelial cells. Proc Natl Acad Sci U S A. 2003. 100:11618–11623.
14. Hong DH, Petrovics G, Anderson WB, Forstner J, Forstner G. Induction of mucin gene expression in human colonic cell lines by PMA is dependent on PKC-epsilon. Am J Physiol. 1999. 277(5 Pt 1):G1041–G1047.
15. Hewson CA, Edbrooke MR, Johnston SL. PMA induces the MUC5AC respiratory mucin in human bronchial epithelial cells, via PKC, EGF/TGF-alpha, Ras/Raf, MEK, ERK and Sp1-dependent mechanisms. J Mol Biol. 2004. 344:683–695.
16. Park SJ, Kang SY, Kim NS, Kim HM. Phosphatidylinositol 3-kinase regulates PMA-induced differentiation and superoxide production in HL-60 cells. Immunopharmacol Immunotoxicol. 2002. 24:211–226.
17. Takeyama K, Dabbagh K, Shim JJ, Dao-Pick T, Ueki IF, Nadel JA. Oxidative stress causes mucin synthesis via transactivation of epidermal growth factor receptor: role of neutrophils. J Immunol. 2000. 164:1546–1552.
Full Text Links
  • TRD
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
Copyright © 2024 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr