Eczematous or atopic dermatitis (AD) is a chronic autoinflammatory skin disorder distinguished by severe itching, scratching, and erosion. Bosentan is an endothelin receptor antagonist with improved immunomodulatory and anti-inflammatory actions. This study aimed to assess the efficiency of topical bosentan in alleviating a 2, 4-dinitrochlorobenzene (DNCB)-induced mouse model of AD. 50 Swiss albino mice were haphazardly grouped into 5 teams of 10 each. The first week of the experiment involved DNCB sensitization on back skin mice, preceding a four-week DNCB challenge to induce AD-like skin inflammation. The control group gets no treatment. The induction group administered DNCB only. Starting two hours after the second sensitization, the vehicle group received topical vehicle solution, the bosentan group received 5% bosentan ointment, and the tacrolimus group received 0.1% tacrolimus ointment once daily for a period of four weeks. Topical bosentan markedly mitigated DNCB-aggravated AD-like skin lesions, as displayed by decreased total dermatitis scores and lowering the upregulated counts of total leukocytes, neutrophils, lymphocytes, monocytes, and eosinophils. Additionally, bosentan dramatically alleviated interleukin (IL)-4 and IL-13 immunohistochemistry scores, as well as IL-1β, IL-6, IL-17, TNFα, and IFN-γ epidermal levels. In conclusion, Bosentan treatment also significantly diminished levels of immunoglobulin E (IgE) and oxidative biomarker malondialdehyde (MDA) and histopathology scores, notably epidermal thickness and inflammation. Bosentan mitigates the severity of DNCB-induced AD-like skin inflammation, possibly owing to its potent anti-inflammatory and immunomodulatory properties.