This study proposes to explore the impact of BTX-A on BPD and the functional mechanism. In this study, RWPE-1 cells were stimulated with 10 ng/mL DHT, followed by treatment with 5, 10, and 20 U/mL BTX-A for 24 h. Significantly increased cell viability, repressed apoptosis, upregulated Vimentin, N-cadherin, and FN, and downregulated E-cadherin were observed in DHT-stimulated RWPE-1 cells, which were drastically rescued by BTX-A treatment. A BPH rat model was established, followed by treatment with 2, 60, and 100 U/mL BTX-A. An increased PW/BW ratio, elevated serum IP: 2038 10920 On Wed 03 May 2023 10:21:56 DHT levels, severe pathological changes in the prostattissue, and failitated EMT progression were observed in BPH Copyright: Ame rcan Scien ific Publishers rats, which were drastically abolished by BTX-A. FurtherDeliveredmore, byin ngentDHT-ha ndled RWPE-1 cells and prostate tissue of BPH rats, HIF-1a and VEGF were significantly upregulated and were greatly downregulated by BTX-A treatment. Finally, the impact of BTX-A on the proliferation and apoptosis of DHT-treated RWPE-1 cells, EMT progression, and VEGF expression was significantly abolished by the overexpression of HIF-1a. Our data revealed that BTX-A alleviated BPH by regulating the progression of EMT via inhibition of HIF-1a.