Hesperidin ameliorates bleomycin-induced experimental pulmonary fibrosis via inhibition of TGF-beta1/Smad3/AMPK and IkappaBalpha/NF-kappaB pathways

Bleomycin (BLM) is a chemotherapeutic agent which is associated with Idiopathic pulmonary fibrosis (IPF) due to its chronic administration. Hesperidin, a bioflavonoid has been reported to possess antioxidant, anti-inflammatory, wound healing, and antiapoptotic potential. To evaluate the therapeutic potential of hesperidin against BLM-induced pulmonary fibrosis and decipher its possible mechanism of action. Intraperitoneal administration of BLM (6 IU/kg) caused induction of IPF in Sprague-Dawley rats. Rats were treated with hesperidin (25, 50, and 100 mg/kg, p.o.) for 28 days, followed by estimation of various parameters in bronchoalveolar lavage fluid (BALF) and lung. Hesperidin (50 and 100 mg/kg) administration significantly ameliorated (p < 0.05) alterations induced by BLM in lung index, percent oxygen saturation, serum ALP and LDH levels, BALF differential cell count, and lung function test. Elevated levels of oxido-nitrosative stress, hydroxyproline, and myeloperoxidase levels in BALF and lung were significantly decreased by hesperidin on day 14. Hesperidin significantly inhibited BLM-induced down-regulated lung Nrf2 and HO-1 as well as up-regulated TNF-α, IL-1β, IL-6, collagen-1, TGF-β, and Smad-3 mRNA expressions. Western blot analysis showed that alteration in lung NF-κB, IκBα, AMPK, and PP2C-α protein expressions were ameliorated by hesperidin on day 28. Furthermore, BLM induced histological and ultrastructural aberrations in the lung which were attenuated by hesperidin treatment. Hesperidin alleviates BLM-induced IPF via inhibition of TGF-β1/Smad3/AMPK and IκBα/NF-κB pathways which in turn ameliorate the modulation of oxido-inflammatory markers (Nrf2 and HO-1) and pro-inflammatory markers (TNF-α, IL-1β, and IL-6) to reduce collagen deposition during pulmonary fibrosis. See also Figure 1(Fig. 1). Open in a separate window Figure 1 Graphical abstract Keywords: AMPK, bleomycin, hesperidin, IkappaBalpha, NF-kappaB, Nrf2, pulmonary fibrosis, Smad3, TGF-beta1 Abbreviations Adenosine monophosphate-activated protein kinase (AMPK), Alkaline Phosphatase (ALP), Bleomycin (BLM), Bronchoalveolar Lavage Fluid (BALF), Enhanced Pause (Penh), Expired Volume (EV), Frequency of breathing (f), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), Glutathione (GSH), Heme oxygenase 1 (HO-1), Hydroxyproline (HP), Idiopathic pulmonary fibrosis (IPF), Interleukins (IL's), Lactate Dehydrogenase (LDH), Malondialdehyde (MDA), Mothers against decapentaplegic homolog-3 (Smad-3), Myeloperoxidase (MPO), Nitric Oxide (NO), Non-phosphorylated AMPK (PP2C-α), Nuclear factor E2-related factor 2 (Nrf2), Nuclear factor-Kappa B (NF-κB), Nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor-alpha (IκBα), Transforming Growth Factor-β (TGF-β), Transmission Electron Microscopy (TEM), Tumor Necrosis Factor-alpha (TNF-α).