An Adaptogen: Withaferin A Ameliorates in Vitro and in Vivo Pulmonary Fibrosis by Modulating the Interplay of Fibrotic, Matricelluar Proteins, and Cytokines

Abstract

Pulmonary fibrosis (PF) is chronic lung disease with only two FDA approved clinically available drugs, with limited safety profile. Inadequate therapy motivated us to explore the effect of vimentin inhibitor Withaferin A, as an anti-fibrotic agent against TGF-β1-induced in vitro fibrotic events and Bleomycin induced in vivo fibrosis with an emphasis on epithelial to mesenchymal transition (EMT), extracellular matrix deposition (ECM), inflammation, and angiogenesis. In vitro EMT and fibrotic events were induced by TGF-β1 in alveolar epithelial cells and human fetal lung fibroblasts followed by treatment with Withaferin A (0.25, 0.5, and 1 μM concentrations) to explore its anti-fibrotic effects. In vivo potential of Withaferin A (2 and 4 mg/kg) was assessed in murine model of Bleomycin induced PF. All the parameters and molecular studies related to PF were performed at the end of treatment period. Withaferin A treatment reduced the progression of PF by modulating the EMT related cell markers both in vivo and in vitro. Withaferin A ameliorated the expression of inflammatory cytokines including NF-κB p65, IL-1β and TNF-α, as well as attenuated the expression of pro-fibrotic proteins including CTGF, collagen 1A2, collagen 3A1, and fibronectin. Expression of angiogenic factors like VEGF, FAK, p38 MAPK, and PLC-γ1 were also inhibited by Withaferin A. Phosphorylation of Smad 2/3 induced by TGF-β1 and Bleomycin were significantly inhibited. Withaferin A suppressed expression of pro-inflammatory, pro-fibrotic, and pro-angiogenic mediators and also reduced the ECM deposition. In a nutshell, Withaferin A could probably prove as an efficient and potential therapeutic against PF.

Withaferin A, a novel compound of Indian ginseng (Withania somnifera), ameliorates Cerulein‐induced acute pancreatitis: Possible role of oxidative stress and inflammation

“…Withaferin A (WFA) is a steroidal lactone, obtained from the roots of Withania somnifera very commonly known as Ashwagandha in Indian Ayurvedic system, and is found to have good anti‐inflammatory activity, due to which it was proved effective in many acute and chronic inflammatory disorders (Berghe, Sabbe, Kaileh, Haegeman, & Heyninck, ; Tekula, Khurana, Anchi, & Godugu, ). In addition, evidence of analgesic, antipyretic, anticancer, antifibrotic, and antibiotic effects of the constituent is worth probing in multiple other disease conditions (Bale, Venkatesh, Sunkoju, & Godugu, ). Anti‐inflammatory activity of WFA is because of downregulation of key transcription factor for the release of inflammatory cytokines such as NF‐κB (Ethridge et al, ).…”

Withaferin A ameliorates renal injury due to its potent effect on inflammatory signaling

“…The observations were found to be in correlation with earlier published reports stating the antioxidant potential of WfA in other organs. 21,37 This observation indicates that the antioxidant activity of WfA is responsible, in part, for the observed nephroprotective activity.…”

“…Interestingly, with our pharmacological intervention, WfA suppressed the levels of the lipid peroxidation products and improved the levels of GSH in obstructed kidneys. The observations were found to be in correlation with earlier published reports stating the antioxidant potential of WfA in other organs . This observation indicates that the antioxidant activity of WfA is responsible, in part, for the observed nephroprotective activity.…”

“…The multiplex analysis of TGF‐β and cytokine/chemokine pathways was performed according to manufacturer's protocol. Kidney tissues were used to detect changes in the TGF‐β2 receptor, phosphorylated Smad2 (Ser465/Ser467), Smad3 (Ser423/Ser425), Akt (Ser473/Ser475), ERK1/2(Thr185/Thr187), and total Smad4 using the Luminex system . Furthermore, a total of 15 types of cytokines/chemokines were estimated in kidney lysates of different groups.…”

“…Kidney tissues were used to detect changes in the TGF-β2 receptor, phosphorylated Smad2 (Ser465/Ser467), Smad3 (Ser423/ Ser425), Akt (Ser473/Ser475), ERK1/2(Thr185/Thr187), and total Smad4 using the Luminex system. 21 Furthermore, a total of 15 types of cytokines/chemokines were estimated in kidney lysates of different groups. The bead-based analysis was carried out in a blind fashion, and the results were expressed in the form of mean fluorescence intensity.…”