Effect of ethyl acetate extract of the whole plant Clerodendrum phlomidis on improving bleomycin (BLM)-induced idiopathic pulmonary fibrosis (IPF) in Rats: In vitro and in vivo research

Idiopathic pulmonary fibrosis (IPF) is a prevalent chronic lung condition of unknown etiology characterized by fibrosis and inflammation. Lung scarring progresses owing to cytokines and immune cells that promote inflammation and fibrosis in idiopathic pulmonary fibrosis (IPF). The anti-inflammatory and anti-fibrotic properties of the ethyl acetate extract of Clerodendrum phlomidis (CPEA), derived from the Indian plant “agnimantha,” are recognized in traditional Ayurvedic medicine. This study investigated the potential protective mechanisms of Clerodendrum phlomidis (CPEA), which have not been previously examined, and demonstrated how CPEA affects bleomycin (BLM)-induced lung fibrosis. Phytometabolomic analysis of Clerodendrum phlomidis was performed using UPLC-ESI-Q/TOF-MS. Free radical scavenging assays were also used to evaluate the antioxidant capacity of the plants using ABTS, DPPH, FRAP, and NO assays. Using ELISA and Griess reagent assays, we assessed the anti-inflammatory effects of CPEA in LPS-induced Jurkat, THP-1, and LL-29 cell lines. This study compared intratracheal injection of BLM-induced IPF in Wistar rats with oral administration of CPEA extract for its anti-fibrotic and anti-inflammatory properties. Multiple techniques were employed, including enzyme-linked immunosorbent assay (ELISA), hydroxyproline, histopathological, biochemical, antioxidant enzyme profiling, and hematological analyses. Polyphenolic compounds were identified using qualitative CPEA. Plant extracts demonstrated free radical-scavenging activity in vitro and exhibited antioxidant properties. CPEA extract reduced TNF-α, IL-1β, and NO levels in LPS-stimulated Jurkat, THP-1, and LL-29 cells. In response to BLM-induced lung and serum conditions in Wistar rats, the CPEA extract significantly reduced (p < 0.05) markers of inflammation and fibrosis (ALP, LDH, TNF-α, CXCL8-MIP2, MMP7, SP-A, SP-D, NO, TBARS, and MPO) and significantly restored antioxidant enzymes (p < 0.05) (GSH, GPx, and GST) and anti-inflammatory cytokines (IL10). Oral CPEA extract attenuates fibrosis, inflammation, oxidative stress, nitrosative stress, and lipid peroxidation in BLM-induced idiopathic pulmonary fibrosis (IPF). CPEA extract improved lung function and increased survival rates. Clinical trials are necessary, as this study indicated that the dietary flavonoid-rich component of CPEA extracts possesses anti-inflammatory and antioxidant properties. CPEA extract restored antioxidant enzyme levels and exerted anti-fibrotic and anti-inflammatory effects in rats with idiopathic lung fibrosis induced by BLM. CPEAs protect against lipopolysaccharide (LPS)-induced inflammation in vitro and bleomycin-induced idiopathic pulmonary fibrosis (IPF) in vivo. The findings of our investigation indicate that CPEA demonstrates therapeutic potential for IPF in human subjects, as evidenced by its capacity to enhance antioxidant, anti-inflammatory, and anti-fibrotic markers in preclinical disease models.