Balanophora involucrata alleviates diabetic nephropathy by inhibiting ferroptosis, modulating serum metabolites and gut microbiota.

Terpenoids and flavonoids were identified as the main components of Balanophora involucrata by UPLC-MS/MS analysis.

• UPLC-MS/MS was employed to identify the main ingredients of BI
• The two primary chemical classes detected were terpenoids and flavonoids
• This chemical profiling informed the mechanistic investigations into BI's therapeutic activity

Balanophora involucrata treatment reduced fasting blood glucose and corrected lipid metabolism disorders in db/db DN mice.

• The DN model was established using C57BLKS/J db/db mice
• BI treatment resulted in reduced fasting blood glucose levels compared to untreated DN mice
• Lipid metabolism disorders were corrected by BI in DN mice
• Serum and urine parameters were assessed to confirm metabolic improvements

Balanophora involucrata ameliorated renal injury in diabetic nephropathy mice.

• Renal histology and ultrastructure were assessed following BI treatment
• Improvements in renal injury markers were observed in BI-treated DN mice
• Both serum/urine parameters and histological assessments confirmed renal protection
• The DN model used C57BLKS/J db/db mice to simulate human diabetic nephropathy

Balanophora involucrata modulated serum metabolites and remodeled gut microbiota composition in DN mice.

• Intrinsic microbial composition was analyzed via 16S rDNA gene sequencing
• Nontargeted metabolomics was used to analyze corresponding serum metabolites
• BI regulated serum metabolites in DN mice
• BI remodeled gut microbiota in DN mice
• These findings suggest a gut microbiota-metabolite axis as part of BI's mechanism of action

Balanophora involucrata inhibited ferroptosis by activating anti-ferroptotic proteins and suppressing pro-ferroptotic proteins.

• Nrf2, GPX4, FPN1, FTH1, and SLC7A11 were activated by BI treatment
• TFR1 and ACSL4 were suppressed by BI treatment
• Protein levels were analyzed by Western blotting
• Lipid peroxidation was evaluated using the Bodipy 581/591 C11 fluorescence assay
• MDA and other ferroptosis-related indicators were detected using commercial kits

BI-containing drug serum protected HK-2 cells from high glucose-induced ferroptosis in vitro.

• HK-2 cells were treated with high glucose to simulate DN conditions
• Cells were then treated with BI-containing drug serum and ferrostatin-1 (fer-1) as a ferroptosis inhibitor comparator
• Cell viability was assessed by CCK-8 assay
• Lipid peroxidation was evaluated using the Bodipy 581/591 C11 fluorescence assay
• Results supported BI's role in inhibiting ferroptosis under diabetic conditions