Keluoxin inhibits the progression of diabetic kidney disease via TNF-α/IKKβ/IκBα/NF-κB p65 signaling pathway

Background: Previous clinical studies have indicated that Keluoxin can be used for the treatment of diabetic kidney disease (DKD). However, the specific mechanisms involved and the primary effective components have not been fully explored. Purpose: To investigate the protective mechanisms of Keluoxin and its main components against DKD. Methods: The animal models included db/db mice treated with Keluoxin or saline, and db/m mice treated with saline. Astragaloside IV (AS-IV) was used to treat HK-2 cells for in vitro experiments. To assess the effects of Keluoxin on DKD, serum and tissue histopathological staining were examined. Western blotting and quantitative real-time PCR (qRT-PCR) were employed to detect the expression levels of fibrosis and inflammation markers. RNA sequencing was conducted on the samples, and downstream mechanisms were validated at the level of animal tissues and cells. Results: Keluoxin significantly reduced blood glucose and urinary microalbumin in db/db mice, alleviated renal tubular damage, inflammation and fibrosis in DKD mice, and did not cause any damage to liver function. Sequencing results indicated that Keluoxin modulated the TNF signaling pathway, NF-κB signal pathway and inflammatory responses. In vivo experiments showed that Keluoxin inhibits the TNF-α/IKKβ/IκBα/NF-κB p65 pathway, thereby providing protection against DKD. Immunoinfiltration analysis indicated that Keluoxin reduced neutrophil infiltration in the renal cortex of DKD mice. Through pharmacological component analysis, we determined that Astragaloside IV (AS-IV) is the main component of Keluoxin. In vitro experiments showed that AS-IV also alleviated the inflammatory response and fibrosis, inhibiting the expression of the TNF-α/IKKβ/IκBα/NF-κB p65 signaling pathway. Conclusion: Keluoxin exerts renal protective effects by inhibiting TNF-α/IKKβ/IκBα/NF-κB p65 signaling pathway with AS-IV being the primary active component.