Background: The antidiabetic effects of the Pleurotus fossulatus on liver and kidney were unexplored. The present study assessed the vital effects of P. fossulatus aqueous extract (PFA-extract) on liver and kidney function in diabetic rats model. Methods: The albino Wistar rats were divided into five groups with six animals in each. Group, I, II, and III were normal, diabetic, and drug control, respectively, and groups IV and V were test groups. As treatment dose, group II received saline, and group III (drug control) received metformin in 100 mg/kg of body weight as a standard drug. Whereas, group IV (D1) and V (D2) were test groups, received PFA-extract in 250 mg/kg, and 500 mg/kg of body weight, respectively. The changes in body weight, blood glucose level (BGL), lipid profile, liver, and kidney biochemical parameters were evaluated. Results: The PFA-extract dose at 500 mg/kg in D2 groups showed very good effects. The body weight was recovered by 97.9% and blood glucose level (BGL) was reduced to 53% in the D2 group. For the lipid profile, total cholesterol (TC), triglyceride (TG), and high-density lipoprotein (HDL) were estimated in blood plasma and their values were 92.31±3.788, 80.85 ±8.962, and 31.35±1.781, respectively. PFA-extract improved the liver function for which aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) were evaluated and recorded to their normal range in D2 group which were 132.01 ±5.553, 58.63±4.390, and 143.26±2.423, respectively. For the kidney function test, creatinine (CRT), blood urea nitrogen (BUN), and uric acid were evaluated and in the D2 group, it was significantly reduced ie, 0.656±0.0707, 15.13±1.463, and 6.27±0.325, respectively. Conclusion: These attributes of PFA-extract make it a potential natural agent to provide protection to the liver and kidney and also reduce the BGL and control the total lipid in type 1 diabetes condition.