Abstract Background: Rhabdomyolysis is a primary skeletal muscle disruption syndrome with circulatory leakage of its intracellular contents and is seriously complicated by acute kidney injury (AKI). Asprosin is a novel glucogenic adipokine expressed in several tissues, including the kidneys, and has been implicated in some renal disorders via many pathogenic mechanisms. Methods: 32 rats were divided equally into the control group, the calcitriol-treated group, the glycerol-treated group, and the glycerol+calcitriol-treated group. Blood, urine, and renal tissue samples were collected for biochemical, histological, immunohistochemical, and gene investigations. Results: Rats given glycerol had elevated levels of asprosin, creatine kinase, creatinine, BUN, renal MDA, IL-6, caspase-3, and caspase-9, as well as PKA mRNA, TGF-β1, and SMAD-3. While creatinine clearance, renal SOD, and catalase were decreased. Marked histopathological changes imply sever renal injury, faint PAS-positive reaction, strong positive immunoreaction for iNOS and TGF-β were found. These changes were reversed in glycerol+calcitriol-treated rats. Asprosin positively correlated with MDA, IL-6, caspase-3, caspase-9, mRNA levels of PKA, TGF-β1and SMAD-3, while it negatively correlated with SOD, and catalase. Conclusion: Serum asprosin levels are increased in rhabdomyolysis-induced AKI and calcitriol protect against AKI via suppressing asprosin and its dependent PKA-TGF-β1-SMAD-3 signaling pathway. Keywords: Rhabdomyolysis; asprosin; acute kidney injury; TGF-β1