<b><i>Introduction:</i></b> Alopecia areata (AA) is a common autoimmune condition that affects anagen hair follicles. The most commonly recognized theory is that it is a T-cell-mediated autoimmune disorder in a genetically susceptible individual. MicroRNAs (miRNAs) and long noncoding RNAs (lncRNAs) were thought to play a function in the pathogenesis. The expression of lncRNA HOTAIR and miRNA-205 and their relation to transforming growth factor β1 (TGF-β1) in AA were not studied. <b><i>Aim:</i></b> The aim of the studywas to evaluate the role of miRNA-205, lncRNA, HOTAIR, and TGF-β1 levels in AA pathogenesis, clinical course, and severity of AA. <b><i>Methods:</i></b> Two groups of subjects were included in this case-control study: 50 patients with AA and 50 healthy matched controls. miRNA-205 and lncRNA HOTAIR expression levels were assayed using quantitative RT-PCR, while serum levels of TGF-β1 were assayed using ELISA techniques. <b><i>Results:</i></b> The serum expression of lncRNA HOTAIR was significantly downregulated in AA patients with a <i>p</i> value &#x3c; 0.001, while the serum expression of both miRNA-205 and TGF-β1 were significantly upregulated in patients. <b><i>Discussion/Conclusion:</i></b> This study highlights the potential role of high serum expression of miRNA-205 and TGF-β1 and the low serum expression of lncRNA HOTAIR in AA pathogenesis. This could be used as a therapeutic target to treat AA.