Modulating HIF-1α/HIF-2α homeostasis with Shen-Qi-Huo-Xue formula alleviates tubular ferroptosis and epithelial-mesenchymal transition in diabetic kidney disease

Ethnopharmacological relevance

Diabetic kidney disease (DKD) is one of the main types of chronic kidney disease, which seriously affects the quality of life of patients. Shen-Qi-Huo-Xue formula (SQHXF), based on the Shen-Qi-Di-Huang decoction, is a traditional Chinese medicine formula for DKD. This study explored the mechanism of action of SQHXF on DKD through analysis of drug components, in vivo and in vitro experiments.

Aim of the study

To elucidate the regulatory mechanisms of HIF-1α/HIF-2α homeostasis on ferroptosis and epithelial-mesenchymal transition (EMT) in renal tubular epithelial cells and the mechanism of action of SQHXF against DKD.

Methods

The components of SQHXF were analyzed using UPLC-Q Exactive HF/MS. The effects of SQHXF on renal function, urinary proteins, glucose-lipid metabolism, hepatic function, renal tissue hypoxia, ferroptosis and EMT were analyzed following gavage of DKD model mice with different SQHXF doses. The effects of changes in HIF-1α and HIF-2α expression on ferroptosis and EMT, as well as the modulatory effects of SQHXF-containing serum, were assessed in vitro. The potential feedback mechanism of HIFs/ferroptosis/EMT was elucidated using HIF-1α knockdown and a ferroptosis inhibitor.

Results

One-hundred and fifty compounds in SQHXF were tested for bloodstream entry. In vivo study showed that SQHXF was able to reduce creatinine, uric acid, fasting plasma glucose, 24-h urinary protein, low-density lipoprotein cholesterol, and aspartate aminotransferase levels, up-regulate HIF-1α, down-regulate HIF-2α, reduce ferroptosis, and alleviate renal fibrosis and EMT in tubular epithelial cells. HIF-1α/HIF-2α imbalance promoted ferroptosis and EMT in HK-2 cells, which was attenuated by SQHXF-containing serum. HIF-1α knockdown decreased HIF-2α expression and reduced ferroptosis and EMT. Inhibition of ferroptosis reduced EMT but failed to regulate HIF-1α and HIF-2α.

Conclusions

SQHXF alleviated ferroptosis and EMT, improved liver and kidney function, reduced proteinuria, and alleviated renal lesions by maintaining equilibrium between HIF-1α and HIF-2α.