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Heme oxygenase-1 (HO-1), which could be highly induced under the stimulation of oxidative stress, functions in reducing the damage caused by oxidative stress, and sulforaphane (SFN) is an antioxidant. This study aims to investigate whether <i>HO-1</i> is involved in the repair of oxidative damage induced by oxidized fish oil (OFO) in <i>Litopenaeus vannamei</i> by sulforaphane (SFN). The oxidative stress model of <i>L. vannamei</i> was established by feeding OFO feed (OFO accounts for 6%), and they were divided into the following four groups: control group (injected with dsRNA-EGFP and fed with common feed), dsRNA-<i>HO-1</i> group (dsRNA-<i>HO-1</i>, common feed), dsRNA-<i>HO-1</i> + SFN group (dsRNA-<i>HO-1</i>, supplement 50 mg kg⁻¹ SFN feed), and SFN group (dsRNA-EGFP, supplement 50 mg kg⁻¹ SFN feed). The results showed that the expression level of <i>HO-1</i> in the dsRNA-<i>HO-1</i> + SFN group was significantly increased compared with the dsRNA-<i>HO-1</i> group (<i>p</i> < 0.05). The activities of SOD in muscle and GPX in hepatopancreas and serum of the dsRNA-<i>HO-1</i> group were significantly lower than those of the control group, and MDA content in the dsRNA-<i>HO-1</i> group was the highest among the four groups. However, SFN treatment increased the activities of GPX and SOD in hepatopancreas, muscle, and serum and significantly reduced the content of MDA (<i>p</i> < 0.05). SFN activated <i>HO-1</i>, upregulated the expression of antioxidant-related genes (<i>CAT</i>, <i>SOD</i>, <i>GST</i>, <i>GPX</i>, <i>Trx</i>, <i>HIF-1α</i>, <i>Nrf2</i>, <i>prx 2</i>, <i>Hsp 70</i>), and autophagy genes (<i>ATG 3</i>, <i>ATG 5</i>), and stabilized the expression of apoptosis genes (<i>caspase 2</i>, <i>caspase 3</i>) in the hepatopancreas (<i>p</i> < 0.05). In addition, knocking down <i>HO-1</i> aggravated the vacuolation of hepatopancreas and increased the apoptosis of hepatopancreas, while the supplement of SFN could repair the vacuolation of hepatopancreas and reduce the apoptosis signal. In summary, <i>HO-1</i> is involved in the repair of the oxidative damage induced by OFO in <i>L. vannamei</i> by SFN.