The aim of this work is to investigate the effects of L-AMB (Amphociene) on liver and lung tissues in adult mice, and the role of vitamin D3 in reducing its side effects. Amphotericin B (AMB), a polyene macrolide antibiotic, is used to treat serious fungal infections and leishmaniasis. It may cause increased serum aminotransferase and hyperbilirubinemia due to interference with hepatic cytochrome P450. This study aimed to determine whether vitamin D could improve liver and lung structural changes. We divided the twenty-four adult male mice into the following groups: The first group (G1) received normal saline orally for 28 days. In the second group (GII), Liposomal amphotericin B (L-AMB) (5 mg/kg body weight) was given for 28 days intravenously. In the group (GIII), vitamin D3 was given daily at a dose of 10000 IU. The fourth group received intravenously L-AMB at a dosage of 5 mg/kg body weight and vitamin D orally for 28 days. Liver sections of group II showed coagulative necrosis and vacuolar degeneration of hepatocytes, as well as Kupffer cell hyperplasia and inflammatory cell infiltration. Group IV showed a normal architecture of liver tissue with mild vacuolation and cloudy swelling of the hepatocytes. The lung sections of the treated group showed interstitial hemorrhages, hemosiderin pigmentation, inflammatory cell infiltration, and accumulation of serous exudate in alveoli. Group IV showed clear alveoli with a few red blood infiltrates. The study found that L-AMB causes structural changes in liver and lung tissue, and treatment with vitamin D3 reduced its deteriorating effects. Monitoring liver and lung function tests in patients with fungal infections who are taking L-AMB for a prolonged period is essential for preventing and treating liver and lung dysfunction.