Pulmonary protective effects of ultrasonic green synthesis of gold nanoparticles mediated by pectin on Methotrexate-induced acute lung injury in lung BEAS-2B, WI-38, CCD-19Lu, IMR-90, MRC-5, and HEL 299 cell lines

Abstract

Regarding applicative, facile, green chemical research, a bio-inspired approach is being reported for the synthesis of Au nanoparticles by pectin (PEC) as a natural reducing and stabilizing agent without using any toxic and harmful reagent under ultrasonic condition. The biosynthesized Au NPs@PEC were characterized by advanced physicochemical techniques like ultraviolet–visible (UV–Vis), Fourier Transformed Infrared spectroscopy (FT-IR), Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), Energy Dispersive X-ray spectroscopy (EDX), and X-ray Diffraction (XRD) study. It has been established that pectin-stabilized Au nanoparticles have a spherical shape with a mean diameter from 5 to 10 nm. In the medicinal part of the present research, the lung BEAS-2B, WI-38, CCD-19Lu, IMR-90, MRC-5, and HEL 299 cell viability was determined by trypan blue assay. The caspase activity colorimetric assay kit and Rhodamine123 fluorescence dye were used to determine the caspase-3 activity and mitochondrial membrane potential, respectively. Apoptosis and DNA fragmentation were determined by the TUNEL test. Also, the inflammatory cytokines concentrations were evaluated by the Rat inflammatory cytokine assay kit. Au NPs@PEC-treated cell cutlers decreased significantly (p ≤ 0.01) the caspase-3 activity, inflammatory cytokines concentrations, and DNA fragmentation, and enhanced the mitochondrial membrane potential and cell viability in the high concentration of Methotrexate-treated lung BEAS-2B, WI-38, CCD-19Lu, IMR-90, MRC-5, and HEL 299 cells. In the antioxidant test, the IC50 of Au NPs@PEC nanocomposite and BHT against DPPH free radicals were 203 and 181 µg/mL, respectively. Finally, Au NPs@PEC may be used as a pulmonary protective supplement to treat acute lung injury.

Keywords

• Pectin
• Gold nanoparticles
• DNA fragmentation
• Apoptosis
• Antioxidant
• Acute lung injury