Tobacco stems are an important part of the tobacco leaf, containing a lot of cellulose, result in the low utilization of tobacco leaves in cigarettes. The pre-treatment technology of tobacco stems is a key to increase the utilization of tobacco stems. In this work, steam explosion treatment was carried out on tobacco stems with different moisture contents containing 2%, 5% and 10%, respectively. The steam explosion is carried out under different pressure and pressure holding time conditions, the changes in physical structure, chemical composition and pyrolysis volatiles of tobacco stem was investigated. The results indicate that steam explosion has a negligible impact on the crystalline structure of cellulose and low-order polysaccharide crystals, while it has an obvious effect on KCl. Steam explosion increases the specific surface area by destroying the amorphous structure on tobacco stems. The contents of chemical constituents and pyrolytic volatiles indicate that there is a significant leaching effect of lignin, but no change in cellulose content. Soluble polysaccharides, the main component of hemicellulose, were converted into pyrolytic volatiles e.g., furfural and furfuryl alcohol. In conclusion, the technology of steam explosion promotes the leaching, decomposition and chemical transformation of amorphous structures on tobacco stems, thus destroying the structure of the cell walls.