Chronic kidney disease (CKD) is considered one of the major public health problems associated with high prevalence, morbidity, and mortality. There is no definite cure for CKD, however studying its underlying mechanisms has provided more understanding of its pathophysiology, hence the development of new therapeutic approaches. Wnt/β-catenin signaling is one of those crucial underlying mechanisms. Tumor necrosis factor-α (TNF-α) was reported to inhibit klotho, reactivate β-catenin and cause tubular cell injury in vitro. Therefore, we studied the effect of infliximab, a monoclonal antibody against TNF-α, on renal Wnt/β-catenin signaling, its downstream target genes and renal injury induced in rats by doxorubicin (DOX). Also, we studied the possible inhibitory effect of pachymic acid on renal Wnt/β-catenin signaling and renal injury in vivo. Rats injected by DOX showed an increased serum cystatin-C, urine albumin/creatinine ratio (UACR), but depleted renal podocin content. DOX markedly increased renal contents of TNF-α, interleukin-6, interleukin-1β. DOX decreased the renal expression of klotho which in turn increased Wnt1, active β-catenin/total β-catenin ratio in renal tissue. Significant increase in renal gene expression of RENIN, ACE, and AT1 was observed. Moreover, renal fibronectin and collagen deposition increased in renal tissue. Treatment with either infliximab, pachymic acid, or/and losartan significantly improved renal function, inhibited inflammatory cytokines and fibrosis. Renal TNF-α was negatively correlated with renal klotho. On the hand, it was positively correlated with renal Wnt1 and active β-catenin/total β-catenin ratio. The combined treatments (infliximab + losartan), (pachymic acid + losartan) were the most effective in improving all studied parameters. In conclusion, this study proved, for the first time, the inhibitory effect of both infliximab and pachymic acid on renal Wnt/β-catenin signaling in DOX-induced nephropathy in vivo by up-regulating renal klotho. Therefore, these results suggest a new role for infliximab and pachymic acid in chronic kidney disease via targeting renal Wnt/β-catenin/renin angiotensin axis.