This study was conducted to investigate the role of endogenous tonicity responsive enhancer binding proteins (TonEBP) on normal physiology of male albino wistar rats and the possible toxic effects when this transcription factor or its transduction products are inhibited. This study was conducted on thirty six male albino rats weighing between 200-250 g, divided randomly into six groups. Rats in 1 st group were used as a control, the 2 nd group of rats received lithium chloride (3mEq/kg, O) every alternative day orally for 2 weeks. 3 rd and 4 th group of rats received a total dose of rottlerin (0.2mg/kg, I/P) and (0.5mg/kg, I/P) respectively every alternative day for 2 weeks, while 5 th and 6 th group rats received both lithium chloride and rottlerin (0.2mg/kg, I/P and 0.5mg/kg, I/P) respectively. The mean arterial pressure was analyzed followed by collection of blood and heart tissue for the estimation of various biochemical parameters such as amino acids, aldose reductase enzyme activity and malondialdehyde levels and kidney for histopathological observations. The study reveals that rottlerin induced inhibition of TonEBP causes deleterious effect on vascular as well as biochemical parameters in both the doses as there was increase in mean arterial pressure, significant reduction in osmoprotective amino acids like taurine, glutamate, phenylalanine and a significant increase in lipid peroxidation in heart tissue. The histopathological observations also revealed degeneration such as mild glomerular, peritubular and blood vessel congestion, presence of inflammatory cells and deposition of colloids. These finding suggests the role of TonEBP in maintenance of normal physiology during osmotic stress in experimental animals.