Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Groiss, S.
Integrative multi-component approaches in mucociliary clearance impaired diseases
Doktoratsstudium der Medizinischen Wissenschaft; Humanmedizin; [ Dissertation ] Medizinische Universität Graz; 2021. pp.
- Autor*innen der Med Uni Graz:
- Groiss Silvia
- Betreuer*innen:
- Brislinger Dagmar
- Litscher Gerhard
- Ratzenhofer-Komenda Beatrice
- Altmetrics:
- Abstract:
- Chronic respiratory diseases (CRDs) are a leading cause of global morbidity and claim about 4 million deaths each year. Limited effectiveness of current therapeutics and the lack of new treatment perspectives turns research towards traditional herbal medication that is believed to offer novel and comprehensive treatment options for the muco-obstructive conditions observed in CRDs. The traditional formulation Bei Mu Gua Lou San (BMGLS) is known for its moisturizing, mu- colytic and expectorant properties over the last two millennia, but the underlying mechanisms remain largely unknown. In this thesis, the effects of BMGLS on mucus-modulation were inves- tigated using an air-liquid-interface (ALI) cell culture system of the respiratory epithelium (REp) reconstituted with primary normal human bronchial epithelial (NHBE) cells. Pioneering steps towards accurate assessment of the effects of BMGLS on NHBE included the development of (i) a protocol for NHBE isolation from tracheal specimen and (ii) a methodology to numerically assess mucous properties from histological images. The latter was achieved by establishing a high-throughput CellProfiler pipeline, which successfully validated an 18-fold difference in MUC5AC expression between a cohort of smoker and non-smoker, thereby confirming data reported in literature. BMGLS application to NHBE ALI cultures induced morphological changes that increased the surface area towards herein termed intra-epithelial structures (IES) and contained intra-IES cilia potentially allowing to disgorge intra-IES mucus. Furthermore, BMGLS reduced the mu- cous area considerably at declining density of mucous solids, potentially indicating detachment of the mucous layer from the periciliary layer via increased hydration. This was further evi- denced by severe accumulations of mucus at marginal areas of the sample that were absent in the controls. Despite significant reduction in mRNA expression of MUC5AC and MUC5B, BMGLS increased MUC5AC protein expression overall increasing the MUC5AC/MUC5B ratio 2-fold. However, the surplus of MUC5AC became increasingly dispersed as demonstrated by reduced IF intensity of the mucins potentially mediated by compound exocytosis-like processes revealed in TEM imaging. Additionally, BMGLS interfered with expression of 15-lipoxygenase (ALOX15), a key mucus-stimulating enzyme. These findings suggest that under applied condi- tions, BMGLS stimulates mucus production while promoting mucus hydration overall supporting facilitated mucus ejection. Therefore, this thesis provides first insights into mucus-modulating effects of BMGLS and en- courages further investigations for its use as a complementary treatment option for patients with CRDs.