J Korean Med Sci.  2011 Jun;26(6):778-784. 10.3346/jkms.2011.26.6.778.

Effects of Scutellarin on MUC5AC Mucin Production Induced by Human Neutrophil Elastase or Interleukin 13 on Airway Epithelial Cells

• Jiang DP
• Perelman J
• Kolosov VP
• Zhou XD

• KMID: 1785964
• DOI: http://doi.org/10.3346/jkms.2011.26.6.778

Abstract

Scutellarin is a flavonoid extracted from a traditional Chinese herb, Erigeron breviscapus. The present study investigated the effect of scutellarin on MUC5AC mucin production and the possible mechanism. Human bronchial epithelial 16 (HBE16) cells were pretreated with scutellarin for 60 min, and then exposed to human neutrophil elastase (HNE) or interleukin (IL)-13 for 12 hr. RT-PCR and ELISA were performed to measure the amount of MUC5AC mucin production. The results showed that scutellarin inhibited MUC5AC expression both in mRNA and protein level induced by HNE in a concentration-dependent manner. However, scutellarin failed to inhibit MUC5AC mucin production induced by IL-13. To investigate the intracellular mechanisms associated with the effect of scutellarin on MUC5AC mucin production, western blotting was carried out to examine the phosphorylation of protein kinase C (PKC), signal transducer and activator of transcription 6 (STAT6) and extracellular signal-regulated kinase 1/2 (ERK1/2). The phosphorylation of PKC and ERK1/2 was attenuated after treatment with scutellarin, whereas STAT6 was not significantly affected. Therefore, it is suggested that scutellarin down-regulates MUC5AC mucin production on HBE16 cells via ERK-dependent and PKC-dependent pathways.

Keyword

Scutellarin; MUC5AC; Human Neutrophil Elastase; Interleukin-13; Airway Epithelial Cells

MeSH Terms

Apigenin/chemistry/*pharmacology
Cells, Cultured
Dose-Response Relationship, Drug
Down-Regulation
Epithelial Cells/*drug effects/metabolism
Erigeron/chemistry
Glucuronic Acids/chemistry/*pharmacology
Humans
Interleukin-13/*pharmacology
Leukocyte Elastase/*pharmacology
Mitogen-Activated Protein Kinase 1/metabolism
Mitogen-Activated Protein Kinase 3/metabolism
Mucin 5AC/genetics/*metabolism
Phosphorylation
Protein Kinase C/metabolism
Respiratory Mucosa/drug effects/*metabolism
STAT6 Transcription Factor/metabolism
Signal Transduction

Figure

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  Fig. 1 The structure of scutellarin.

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  Fig. 2 Up-regulation of MUC5AC protein induced by human neutrophil elastase (HNE) on HBE16 cells. Cells were collected after exposed to HNE at different concentrations. The amount of MUC5AC was measured by ELISA. Data represent means ± SEM of four experiments. *P < 0.05 compared with the negative control group.

• 

  Fig. 3 Up-regulation of MUC5AC induced by IL-13 on HBE16 cells. Cells were collected after exposed to IL-13 at different concentrations. The amount of MUC5AC was measured by ELISA. Data represent means ± SEM of four experiments. *P < 0.05 compared with the negative control group.

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  Fig. 4 Effects of scutellarin (scu) on MUC5AC mRNA (A) and protein (B) expression after exposed to HNE. The cells were pretreated with scu or medium only for 60 min before the addition of 0.1 µM/L HNE. Total RNAs were reverse transcribed and used for PCR amplication (A). The amount of MUC5AC protein was measured by ELISA (B). Data represent means ± SEM of four experiments. *P < 0.05 compared with the negative group; †P < 0.05 compared with the HNE control group.

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  Fig. 5 Effects of scutellarin (scu) on MUC5AC mRNA (A) and protein (B) expression after exposed to IL-13. The cells were pretreated with scu or medium only for 60 min before the addition of 10 ng/mL IL-13. Total RNAs were reverse transcribed and used for PCR amplication (A). The amount of MUC5AC protein was measured by ELISA (B). Data represent means ± SEM of four experiments. *P < 0.05 compared with the negative control group; ‡P > 0.05 compared with the IL-13 control group.

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  Fig. 6 Effects of scutellarin (scu) on the phosphorylation of PKC induced by HNE. The cells were pretreated with scu, Calphostin C or medium only for 60 min before the addition of 0.1 µM/L HNE. The phosphorylation of PKC was measured by western. Data represent means ± SEM of four experiments. *P < 0.05 compared with the control group; †P < 0.05 compared with the HNE control group.

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  Fig. 7 Effects of scutellarin (scu) on the phosphorylation of ERK1/2 induced by HNE. The cells were pretreated with scu, PD98059 or medium only for 60 min before the addition of 0.1 µM/L HNE. The phosphorylation of ERK1/2 was measured by western. Data represent means ± SEM of four experiments. *P < 0.05 compared with the negative group; †P < 0.05 compared with the HNE control group.

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  Fig. 8 Effects of scutellarin (scu) on the phosphorylation of STAT6 induced by IL-13. The cells were pretreated with scu, A771726 or medium only for 60 min before the addition of 10 ng/mL IL-13. The phosphorylation of STAT6 was measured by western. Data represent means ± SEM of four experiments. *P < 0.05 compared with the negative group; †P < 0.05 compared with the IL-13 control group; ‡P > 0.05 compared with the IL-13 control group.

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