484-P: Abnormal Ketone Body Metabolism Caused by Bdh1 Deficiency Promoted the Progression of Diabetic Kidney Disease by Activating ROS

Abstract

Objective: This study was intended to reveal the mechanism of ketone body metabolism abnormality caused by bdh1 deficiency promoting the progression of diabetic kidney disease(DKD) by activating reactive oxygen species (ROS). Methods: Eight-week-old wild type(n=8) and db/db (n=8) mice were treated with normal diet and high-fat diet for three months, respectively. Then two groups of mice kidney samples were taken for RNA-Seq analysis to select the target gene. It was verified by qRT-PCR, Western blot, immunofluorescence, and immunohistochemistry in mice kidney. A high glucose-induced ROS activation model of HK-2 renal cells was established, and then siRNA was used to interfere with bdh1. Results: RNA-seq showed that the pathway of ketone body metabolism in DKD changed significantly, and further analysis showed that bdh1, the key protein of ketone body metabolism, decreased significantly. The sequencing result was further confirmed by qRT-PCR, Western blot, immunofluorescence, and immunohistochemistry, and we found that bdh1 was mainly expressed in tubular epithelial cells. In order to identify the molecular mechanism of bdh1 involved in DKD, we constructed the high glucose-induced ROS activation model of HK-2 cells in vitro. It was found that bdh1 was significantly down-regulated in high glucose treated HK-2 cells. The knockdown of bdh1 by siRNA could promote the production of ROS induced by high glucose. Further research found that bdh1 promoted the expression of nrf2 then inhibited ROS through the ketone body metabolism pathway, thus protected HK-2 cells in a high glucose environment. Conclusions: Bdh1 promoted the expression of nrf2 through the pathway of ketone body metabolism and inhibited the production of ROS to protect HK-2 cells in a high glucose environment. Abnormal ketone body metabolism caused by bdh1 deficiency participated in the progress of DKD. Disclosure S. Wan: None. F. Teng: None. Z. Jiang: None. Y. Xu: None.