The Mycoplasma pneumoniae (M.pneumoniae) was accounted to 3-10% of total pneumonia incidences. In recent decades, metallic nanoparticles were extensively examined as nano-antibiotics.In this investigation, we intended to inspect the therapeutic potential of Zinc oxide nanoparticles (ZnONPs) from (Corydalis yanhusuo) C. yanhusuo against the mycoplasma infected pneumonia in mice.The ZnONPs were formulated via green route technique and characterized by UV-vis spectroscopy, transmission electron microscopy, Fourier transform infrared technique, and atomic force microscopy. The antimicrobial activity of formulated ZnONPs was tested by well diffusion method. The total protein, interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor alpha (TNF-α) and transforming growth factor (TGF) status in the BALF of M. pneumonia infected animals were investigated via kit method. The expressions of ERK1/2, JNK1/2, and NF-κB were examined through the Western blotting. The Histopathological analysis of lung tissues of experimental animals was done.The UV-vis spectroscopy and TEM examinations were proved the existence of CY-ZnONPs. The formulated CY-ZnONPs were displayed the potential antimicrobial activity. The supplementation of CY-ZnONPs were noticeably diminished the total protein and IL-6, IL-8, and TNF-α levels in the BALF of pneumonia mice. The ERK1/2, JNK1/2, and NF-κB expressions were appreciably diminished in the CY-ZnONPs supplemented mice. It also reduced the inflammatory cells penetration, and exhibited normal tissue arrangements in the lung tissues of pneumonia mice.The findings of this investigation were proved that the synthesized CY-ZnONPs has the potential to ameliorate the M. pneumoniae infected pneumonia in investigational mice.