Abstract: OBJECTIVE To study the effect of Astragaloside Ⅳ on mitochondrial damage of human peritoneal mesothelial cells (HPMCs) induced by high-glucose peritoneal dialysis and explore its intervention mechanism. METHODS ①HPMCs HMrSV5 were cultured in RPMI 1640 (containing 1% FCS) medium for 24 hours and divided into six groups: blank group: normal culture medium 2 mL; High-glucose peritoneal dialysis group: 4.25% PDS 1 mL + normal culture medium 1 mL; Low-dose Astragaloside Ⅳ group: the final concentration of Astragaloside Ⅳ was 10 μmol/mL; Middle-dose Astragaloside Ⅳ group: The final concentration of Astragaloside Ⅳ was 20 μmol/mL；High-dose Astragaloside Ⅳ group: The final concentration of Astragaloside Ⅳ was 40 μmol/mL； Positive drug control group: pifenidone 50 μmol/mL. ②The proliferative activity of HMrSV5 was measured by CCK-8 method， the level of reactive oxygen species (ROS) was measured by flow cytometry， the level of membrane potential of HMrSV5 was measured by Rhodamine 123 fluorescence staining， the depolarization of mitochondrial membrane potential of HMrSV5 was measured by JC-1 method， and the expression of mitochondrial morphology and function-related proteins of HPMCs was detected by Western blot. RESULTS ①The results of CCK-8 showed that compared with the model group， the OD value and survival rate of HPMCs in Astragaloside Ⅳ treatment group increased significantly， which was concentration-dependent. Astragaloside Ⅳ had protective effect on HPMCs. ②Flow cytometry showed that the mitochondrial ROS production of HPMCs increased after high glucose peritoneal dialysis stimulation， while Astragaloside Ⅳ could effectively reduce the ROS level. ③After JC-1 and Rhodamine 123 staining， flow cytometry showed that with the increase of Astragaloside Ⅳ concentration， the depolarization degree of mitochondrial membrane potential of HPMCs decreased. At the same time， the mitochondrial membrane potential of HPMCs increased effectively， and the permeability of HPMCs mitochondrial membrane decreased. Astragaloside Ⅳ could partly repair the damage of HPMCs mitochondria caused by high glucose peritoneal dialysis. ④Western blot results showed that the expression levels of mitochondrial mitotic proteins DRP1 and FIS1 were significantly decreased， while the expression levels of mitochondrial extracorporeal membrane binding proteins OPA1 and TOM70 were significantly increased. CONCLUSION ①High glucose peritoneal dialysis can induce mitochondrial damage in HPMCs. ②Astragaloside Ⅳ can improve the oxidative stress injury of HPMCs mitochondria induced by high glucose peritoneal dialysate and protect the stability of their structure and function by promoting the expression of OPA1 and TOM70 proteins， reducing the expression levels of DRP1 and FIS1 proteins， reducing the depolarization of mitochondrial membrane potential， effectively increasing the membrane potential and reducing the permeability of mitochondrial membrane.