Background and Purpose The prognosis of cardiorenal dysfunction induced by diabetes mellitus (DM), which belongs to cardiorenal syndrome type 5, is poor and its pathogenesis remains elusive. Here we attempt to clarify whether CX3CL1 might be a therapeutic target for cardiorenal dysfunction in diabetes. Experimental Approach Effects of CX3CL1 on cardiorenal function, myocardial and renal fibrosis, and apoptosis were examined using four types of cultured cardiomyocytes or renal cells and two diabetic mouse models, streptozotocin-induced DM or non-obese diabetic (NOD) mice. The effect of sodium glucose cotransporter 2 inhibitor canagliflozin on CX3CL1 expression was also investigated in vivo or in vitro. Key Results Cardiac and renal CX3CL1 protein levels were significantly increased in both STZ-induced diabetic mice and NOD mice, and that hyperglycemia led to persistent CX3CL1 expression in the heart and kidneys even after it was controlled by insulin. In cultured cardiac and renal cells, soluble CX3CL1 accelerated mitochondrial-dependent apoptosis via activation of the RhoA/ROCK1-Bax pathway and promoted fibrosis through cellular phenotypic trans-differentiation mediated by the TGFβ/Smad pathway. In the two diabetic mouse models, knockout of CX3CL1 receptor CX3CR1 or treatment with an CX3CL1 neutralizing antibody significantly improved cardiorenal dysfunction by inhibiting apoptosis, mitochondrial dysfunction, and fibrosis. Moreover, sodium glucose cotransporter 2 inhibitor canagliflozin significantly downregulated cardiac and renal CX3CL1 expression and improved cardiorenal dysfunction. Conclusions and Implications These findings indicate that CX3CL1 could be a new therapeutic target for diabetes-induced cardiorenal dysfunction.