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Resistant hypertension (RH) remains a major cardiovascular challenge despite optimal pharmacological treatment. Intermittent fasting (IF) has demonstrated beneficial effects in various diseases, but its impact on RH and the underlying mechanisms remain unclear. In this study, we explored the effects of a 2-week IF regimen (16-hour fasting/8-hour eating) on RH patients and spontaneously hypertensive rats (SHRs) resistant to antihypertensive drugs. We found that IF significantly reduced blood pressure in RH patients, accompanied by a shift in the gut microbiota, including increased abundance of Akkermansia muciniphila and Adlercreutzia equolifaciens. These microbiota alterations were correlated with a decrease in lipopolysaccharide (LPS) and trimethylamine-N-oxide (TMAO) levels, and an increase in short-chain fatty acids (SCFAs). Furthermore, fecal microbiota transplantation (FMT) from drug-resistant SHRs successfully transferred both hypertension and impaired drug efficacy to recipient rats. Supplementation with Akkermansia muciniphila and Adlercreutzia equolifaciens significantly lowered blood pressure in SHR rats resistant to antihypertensive drugs. In RH patients, oral supplementation with Akkermansia muciniphila reduced blood pressure and normalized LPS, TMAO, and SCFA levels. Our findings provide both clinical and mechanistic evidence supporting IF and A. muciniphila supplementation as promising non-pharmacological approaches for managing resistant hypertension.