Modulation of COX-2 and NADPH oxidase-4 by alpha-lipoic acid ameliorates busulfan-induced pulmonary injury in rats

Abstract

This study aimed to explore the potential protective effect of α-lipoic acid on busulfan-induced pulmonary fibrosis in rats.Main methods: Eighteen adult male rats were divided into 3 groups; control, busulfan, and busulfan plus α-lipoic acid groups. Lung index ratio, serum level of proinflammatory cytokine were assessed. The activities of antioxidant enzymes and lipid peroxidation products were estimated in the lung tissues in addition to the histopathological analyses. The deposition of the collagen in the lung tissues was evaluated by Sirius red staining. The expressions of α-smooth muscle actin (α-SMA), TNF-α, and Caspase 3 were determined immunohistochemically.The pulmonary expression of COX-2 and NOX-4 mRNA was assessed using qRT-PCR. Key findings: Administration of ALA significantly protect the lung against BUS-induced pulmonary fibrosis, besides the upregulation of antioxidants, and downregulation of pro-inflammatory cytokines. Also, it reduced collagen deposition that associated with a decreased expression of α-SMA, TNF-α, and Caspase 3 in the lung tissues.Moreover, ALA significantly upregulated the expression of COX-2 concomitant with the downregulation of elevated NOX-4. Significance: ALA attenuates the lung cytotoxicity of busulfan through its anti-inflammatory, anti-apoptotic, and antifibrotic effects that may be mediated by upregulation of COX-2 and downregulation of NOX-4.

Methotrexate-Induced Alteration of Renal Aquaporins 1 and 2, Oxidative Stress and Tubular Apoptosis Can Be Attenuated by Omega-3 Fatty Acids Supplementation

“…The immunoperoxidase method was performed on three μm thick renal cortical transverse sections [ 44 , 45 , 46 ]. In brief, the slides were deparaffinized and endogenous peroxidase was blocked.…”

Targeting Hydrogen Sulfide Modulates Dexamethasone-Induced Muscle Atrophy and Microvascular Rarefaction, through Inhibition of NOX4 and Induction of MGF, M2 Macrophages and Endothelial Progenitors

“…Real-time PCR equipment (Pikoreal 96, ThermoScientific) was used to replicate cDNA templates. The amplification process consisted of a 20 μL total volume mixture [ 10 ] μL of HERA SYBR green PCR Master Mix (Willowfort, West Midlands, UK), 2 μL of cDNA template, 2 μL (10 pmol/μL) of each gene primer, and 6 μL of nuclease-free water], and was carried out using the following program: 95 °C for 2 min, followed by 40 cycles of 95 °C for 10 s, and 60 °C for 30 s. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was employed as a control gene [ 25 ], and the sequences of the used primer pairs are listed in Table 2 . Vivantis provided the primer sets (Vivantis Technologies, Malaysia).…”

The effects of lipoic acid on respiratory diseases

“…For instance, LA (20 mg/kg/day i.p. for 6 weeks) significantly decreased the levels of inflammatory cytokines (IL-1β, IL-6, IL-10 and TNF-α), a myofibroblast marker (α-SMA) and caspase-3 in the lungs of rats that were administered busulfan [119] . Previous studies suggested that COX-2-derived prostaglandins could reduce some histological changes in collagen production in models of lung fibrosis [120] , [121] .…”

“…It has been recently shown that NOX-4 is expressed in pulmonary arterial adventitial fibroblasts and subsequently stimulates proliferation and inhibits apoptosis in fibroblasts [122] . Elhadidy et al reported that LA (20 mg/kg BW) decreased the deposition of collagen in busulfan-induced pulmonary fibrosis, which was associated with the upregulation of COX-2 and its downstream effector PGE2, as well as the downregulation of NOX-4 [119] . In another study, LA (50 mg/kg/day fora 28 days) regulated mitochondrial homeostasis and enhanced antioxidant activity to attenuate SiO 2 -induced pulmonary fibrosis by activating the AMPK/PGC1α and NRF2 pathways.…”

“…with 14-day intervals LA 20 mg/kg*day for 6 weeks (i.p.) after the first challenge Attenuating PF (↓hydroxyproline, α-SMA, collagen deposition) by exerting anti-inflammatory (↓TNF-α, IL-1β, IL-6, IL-10) and antioxidant (↓MDA; ↑GPx, SOD) and antiapoptotic (↓caspase-3) effects [119] In vivo Wistar rats Amiodarone 40 mg/kg*day (p.o.) for 4 weeks LA 100 mg/kg* day (p.o.)…”