Curcumin prophylaxis refurbishes alveolar epithelial barrier integrity and alveolar fluid clearance under hypoxia

We have studied the prophylactic efficacy of curcumin to ameliorate the impairment of tight junction protein integrity and fluid clearance in lungs of rats under hypoxia. A549 cells wereexposed to 3 % O₂ for 1 h, 3 h, 6 h, 12 h, 24 h and 48 h and rats were exposed to 7620 m for 6 h. NF-κB, Hif-1α and their related genes, tight junction protein (TJ) (ZO-1, JAM-C, claudin-4 and claudin-5, claudin-18) expressions were determined in A549 cells and lungs of rats by western blotting, ELISA and their activity by reporter gene assay, siRNAp65 knock out. Tissue specific localization of tight junction protein was determined by immunohistochemistry and immunoflorescence. Further transmission electron microscopy (TEM) was used to visualize the TJ structures between pulmonary epithelial cells. Blood gas and hematological parameters were also assessed. Later we checked, whether prior treatment with curcumin can restore the altered alveolar epithelial barrier integrity that is compromised through inflammatory mediators under hypoxia, A549 cells were pre-treated (1 h) with 10 μM curcumin and rats with 50 mg curcumin/kg BW and exposed to hypoxia. Curcumin pre-treatment both in vitro and in vivo showed significant changes in TJ protein integrity, attenuated NF-κB activity with reduced expression of its regulatory genes in lung tissues, serum and bronchoalveolar lavage fluid (BALF) along with stabilized HIF-1α levels under hypoxia. NF-κB inhibitors MG132, SN50 or siRNA mediated p65 knock down significantly reduced the dextran FITC influx into the lungs. The present study indicates that, curcumin prophylaxis augments alveolar epithelial barrier integrity and alveolar fluid clearance under hypoxia.