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Nitric oxide estimation (modified Griess assay)
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Lung tissues were homogenized and centrifuged in ice-cold condition at 12,000 rpm for 12 min and then supernatants were collected and used immediately to measure nitric oxide (NO) levels. Griess reagent was used to evaluate the nitrate levels (NO) of the samples by mixing an equal proportion of both Griess reagent and supernatants of the lung tissue samples, incubated for 10 min in dark at 37 °C, and absorbance was measured at 548 nm. Data obtained from the experiments were normalized using protein content and expressed as μM/mg of tissue.
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