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필용방감길탕이 기도 뮤신의 분비, 생성, 유전자 발현 및 점액 과다 분비에 미치는 영향

Effect of Piryongbanggamgil-tang on Airway Mucin Secretion, Production, Gene Expression and Hypersecretion of Mucus

김장현|민상연|김윤영
대한한방소아과학회 (The Association of Pediatrics of Korean Medicine)|2014.05|대한한방소아과학회지 (The Journal of Pediatrics of Korean Medicine)|제28권 제2호|pp. 56-71

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키워드

MucinGeneAirwayPiryongbanggamgil-tang (PGGT)
주제분류한의학

초록

ObjectivesIn this study, the author tried to investigate whether piryongbang-gamgil-tang (PGGT) significantly affect in vitroairway mucin secretion, PMA- or EGF- or TNF-α-induced MUC5AC mucin production / gene expression from humanairway epithelial cells and increase in airway epithelial mucosubstances and hyperplasia of tracheal goblet cells of rats. Materials and MethodsFor in vitro experiment, confluent RTSE cells were chased for 30 min in the presence of PGGT to assess the effectof PGGT on mucin secretion by enzyme-linked immunosorbent assay (ELISA). Also, effect of PGGT on PMA- or EGForTNF-α-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) wereinvestigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of PGGT and treated with PMA(10 ng/ml) or EGF (25 ng/ml) or TNF-α (0.2 nM) for 24 hrs, to assess both effect of PGGT on PMA- or EGF- or TNF-α-inducedMUC5AC mucin production by ELISA and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). For in vivo experiment, the author induced hypersecretion of airway mucus and goblet cell hyperplasia by exposureof rats to SO2 during 3 weeks. Effect of orally-administered PGGT during 2 weeks on increase in airway epithelialmucosubstances from tracheal goblet cells of rats and hyperplasia of goblet cells were assesed by using histopathologicalanalysis after staining the epithelial tissue with alcian blue. Possible cytotoxicities of PGGT in vitro were assessed by examining LDH release from RTSE cells and the rateof survival and proliferation of NCI-H292 cells. In vivo liver and kidney toxicities of PGGT were evaluated by measuringserum GOT/GPT activities and serum BUN/creatinine concentrations of rats after administering PGGT orally. Results(1) PGGT did not affect in vitro mucin secretion from cultured RTSE cells. (2) PGGT significantly inhibitedPMA-, EGF-, and TNF-α-induced MUC5AC mucin productions and the expression levels of MUC5AC mRNA fromNCI-H292 cells. (3) PGGT decreased the amount of intraepithelial mucosubstances and showed the tendency ofexpectorating airway mucus already produced. (4) PGGT increased LDH release from RTSE cells. However, PGGTdid not show in vivo liver and kidney toxicities and cytotoxicity to NCI-H292 cells. ConclusionThe result from this study suggests that PGGT can regulate the production and gene expression of airway mucinobserved in diverse respiratory diseases accompanied by mucus hypersecretion and do not show in vivo toxicity toliver and kidney functions after oral administration. Effect of PGGT with their components should be further studiedusing animal experimental models that reflect the diverse pathophysiology of respiratory diseases through future investigations.

Abstract

ObjectivesIn this study, the author tried to investigate whether piryongbang-gamgil-tang (PGGT) significantly affect in vitroairway mucin secretion, PMA- or EGF- or TNF-α-induced MUC5AC mucin production / gene expression from humanairway epithelial cells and increase in airway epithelial mucosubstances and hyperplasia of tracheal goblet cells of rats. Materials and MethodsFor in vitro experiment, confluent RTSE cells were chased for 30 min in the presence of PGGT to assess the effectof PGGT on mucin secretion by enzyme-linked immunosorbent assay (ELISA). Also, effect of PGGT on PMA- or EGForTNF-α-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) wereinvestigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of PGGT and treated with PMA(10 ng/ml) or EGF (25 ng/ml) or TNF-α (0.2 nM) for 24 hrs, to assess both effect of PGGT on PMA- or EGF- or TNF-α-inducedMUC5AC mucin production by ELISA and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). For in vivo experiment, the author induced hypersecretion of airway mucus and goblet cell hyperplasia by exposureof rats to SO2 during 3 weeks. Effect of orally-administered PGGT during 2 weeks on increase in airway epithelialmucosubstances from tracheal goblet cells of rats and hyperplasia of goblet cells were assesed by using histopathologicalanalysis after staining the epithelial tissue with alcian blue. Possible cytotoxicities of PGGT in vitro were assessed by examining LDH release from RTSE cells and the rateof survival and proliferation of NCI-H292 cells. In vivo liver and kidney toxicities of PGGT were evaluated by measuringserum GOT/GPT activities and serum BUN/creatinine concentrations of rats after administering PGGT orally. Results(1) PGGT did not affect in vitro mucin secretion from cultured RTSE cells. (2) PGGT significantly inhibitedPMA-, EGF-, and TNF-α-induced MUC5AC mucin productions and the expression levels of MUC5AC mRNA fromNCI-H292 cells. (3) PGGT decreased the amount of intraepithelial mucosubstances and showed the tendency ofexpectorating airway mucus already produced. (4) PGGT increased LDH release from RTSE cells. However, PGGTdid not show in vivo liver and kidney toxicities and cytotoxicity to NCI-H292 cells. ConclusionThe result from this study suggests that PGGT can regulate the production and gene expression of airway mucinobserved in diverse respiratory diseases accompanied by mucus hypersecretion and do not show in vivo toxicity toliver and kidney functions after oral administration. Effect of PGGT with their components should be further studiedusing animal experimental models that reflect the diverse pathophysiology of respiratory diseases through future investigations.

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