Exploring the anti-inflammatory potential of bioactive compounds in inflamed a549 lung cells

Chronic obstructive pulmonary disease (COPD) is a leading cause of morbidity and mortality worldwide, with current treatments primarily focusing on symptom management rather than addressing the underlying inflammatory mechanisms. The nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome plays a pivotal role in COPD pathophysiology by mediating the release of pro-inflammatory cytokines, IL-1β and IL-18, contributing to lung tissue damage and disease progression. Given the limitations of conventional therapies, natural bioactive compounds with antioxidant and anti-inflammatory properties represent promising alternative strategies for COPD management. This thesis investigates the anti-inflammatory potential of natural extracts from Cannabis sativa L. seeds and sprouts, bee pollen, and the microalga Chlorella sp. strain C2, aiming to identify sustainable therapeutic strategies for COPD. The study explores the effects of enriched germination conditions on C. sativa sprouts using Chlorella sp. and the endophytic bacterium Can_S11, known for producing indoleacetic acid. Bee pollen, a functional food with antioxidant and anti-inflammatory properties, is also assessed for its potential to alleviate respiratory inflammation. Additionally, the anti-inflammatory activity of Chlorella sp. strain C2, cultivated under standard and nutrient-deprived conditions, is evaluated. The anti-inflammatory activity of these natural extracts was tested on an A549 lung epithelial cell model inflamed with tumor necrosis factor-alpha (TNF-α). Furthermore, a protocol was established to induce NLRP3 inflammasome activation via lipopolysaccharide (LPS) and ATP stimulation, followed by selective inhibition using INF39 and CY-09. The results demonstrated that C. sativa sprouts grown under enriched conditions exhibited enhanced levels of polyphenols and flavonoids, correlating with significant antioxidant and anti-inflammatory activities. Bee pollen and microalgal extracts also showed promising results in antioxidant activity and reducing pro-inflammatory cytokines. Further experiments on NLRP3 inflammasome activation, using LPS and ATP stimulation, showed that hemp seeds and bee pollen extracts significantly inhibited NLRP3 activation and caspase-1 cleavage, comparable to selective NLRP3 inhibitors CY-09. This research highlights the potential of natural bioactive compounds as complementary therapies in COPD management, offering a natural approach to harnessing plant-derived resources.