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Article Abstract
Background: Doxorubicin (DOX) is a widely used anticancer drug with a marked nephrotoxic effect. The Chinese medicine Tangshen Formula (TSF) has a kidney-protective effect in diabetic kidney disease patients and has been shown to attenuate kidney damage in rodents.
Purpose: Our study explored the putative targets and regulatory mechanisms of TSF in attenuating DOX-induced nephrotoxicity.
Methods: Protein factor microarray, bioinformatics analysis, MPC5 cell line were used in this work. Through the comprehensive means of biological information analysis, in vitro experiments, in vivo experiments and molecular docking, the pathological relationship between TSF and DOX induced nephrotoxic was revealed, and the potential small-molecule inhibitor were identified.
Results: TSF administration significantly alleviated albuminuria and histopathologic lesions. Rat renal cytokine array and GO analysis revealed that signaling pathways of TSF were closely related to apoptosis and pyroptosis. In vivo results verified that TSF inhibited the apoptosis and pyroptosis of DOX induced nephrotoxic rat podocyte. TSF increased levels of WT1, and nephrin, which are podocyte markers, and reduced levels of GSDMD, which is a pyroptosis marker protein in the rat kidney. Furthermore, immunofluorescence co-localization confirmed that TSF could decrease the level of BAX and increase the level of BCL2 in podocytes of DOX induced nephrotoxic rat. In vitro, BTSA1, which is a BAX activator, could mitigate the beneficial effects of TSF. In addition, we identified the compounds with high binding efficiency to BAX from TSF in plasma by CDOCKER, and the top three were selected for verification by western blot, ginsenoside having the best effect to MPC pyroptosis.
Conclusion: This study demonstrated a novel action of TSF in attenuating podocyte death by regulating BAX-mediated crosstalk between pyroptosis and apoptosis. TSF is a potential therapeutic herbal therapy against AN, and ginsenoside is the most effective inhibition of BAX levels.
Chemical Compounds: Ginsenoside Rb1 (PubChem CID: 9898279); Notoginsenoside R1 (PubChem CID: 441934); Naringin (PubChem CID: 442428).
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