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However, its prevention and treatment face significant challenges. This study explored the effects of hydrogen sulfide (H\n                      <jats:sub>2<\/jats:sub>\n                      S) intervention in mice with DM and HLI, while concurrently investigating its regulatory effects on gut microbial homeostasis.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      The diabetic model in C57BL\/6J mice was established through intraperitoneal injection of streptozotocin. The HLI model was created by ligating and severing the femoral artery, with subsequent initiation of a 21-day exogenous H\n                      <jats:sub>2<\/jats:sub>\n                      S intervention. Fecal samples from the mice were collected at four time points: before model establishment, 3 days after successful induction of the diabetes model, 3 days after establishment of the HLI model, and after 21 days of H\n                      <jats:sub>2<\/jats:sub>\n                      S intervention for metagenomic analysis. Body weight, blood glucose levels, and hindlimb blood flow in the mice were monitored. Additionally, functional assessment and histopathological examination of the ischemic skeletal muscle were performed to evaluate contractile and morphological properties.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      H\n                      <jats:sub>2<\/jats:sub>\n                      S administration significantly enhanced hindlimb blood perfusion and restored plasma H\n                      <jats:sub>2<\/jats:sub>\n                      S concentrations in diabetic mice with HLI, concurrently improving both function and morphological integrity of the ischemic skeletal muscle. Bacterial abundance at the phylum level showed changes over the course of the experiment, particularly in\n                      <jats:italic>Bacteroidetes<\/jats:italic>\n                      and\n                      <jats:italic>Firmicutes<\/jats:italic>\n                      . In the DM\u2009+\u2009HLI group, the\n                      <jats:italic>Firmicutes<\/jats:italic>\n                      -to-\n                      <jats:italic>Bacteroidetes<\/jats:italic>\n                      ratio was significantly elevated; however, H\n                      <jats:sub>2<\/jats:sub>\n                      S treatment downregulated this alteration. H\n                      <jats:sub>2<\/jats:sub>\n                      S intervention modulated the abundance of various bacterial species, increasing\n                      <jats:italic>Lactobacillus murinus<\/jats:italic>\n                      and\n                      <jats:italic>Faecalibacterium prausnitzii<\/jats:italic>\n                      , while simultaneously downregulating inflammation-related bacteria such as\n                      <jats:italic>Ruminococcus sp. JE7A12<\/jats:italic>\n                      . Microbial network analysis revealed that the DM\u2009+\u2009HLI and H\n                      <jats:sub>2<\/jats:sub>\n                      S groups had lower network complexity than the control group. Furthermore, functional metagenomic profiling identified 28 differentially expressed genes, which were annotated to 8 primary and 30 secondary KEGG pathways, with 6 genes specifically enriched in carbohydrate metabolism pathways.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      Exogenous H\n                      <jats:sub>2<\/jats:sub>\n                      S administration improved hindlimb blood perfusion, restored contractile function, and preserved morphological integrity of ischemic skeletal muscle in diabetic mice with HLI. Concurrently, H\n                      <jats:sub>2<\/jats:sub>\n                      S treatment altered the abundance of gut microbiota, improving microbial balance. Targeting the gut microbiota via H\u2082S suggests a potential translational avenue that warrants causal investigation for the treatment of diabetic limb ischemia. Further studies are warranted to establish causal relationships and elucidate the underlying mechanisms linking H\n                      <jats:sub>2<\/jats:sub>\n                      S, gut microbiota, and vascular recovery.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12866-026-05167-5","type":"journal-article","created":{"date-parts":[[2026,6,5]],"date-time":"2026-06-05T15:04:37Z","timestamp":1780671877000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The impact of hydrogen sulfide on gut microbiota of diabetic mice with lower limb arterial ischemia"],"prefix":"10.1186","volume":"26","author":[{"given":"Xuan","family":"Qiu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenxiao","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Man","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shibo","family":"Lei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohan","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuxin","family":"Miao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zheng","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuming","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiyong","family":"Hou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,6,5]]},"reference":[{"issue":"9","key":"5167_CR1","doi-asserted-by":"publisher","first-page":"e171","DOI":"10.1161\/CIR.0000000000001005","volume":"144","author":"MH Criqui","year":"2021","unstructured":"Criqui MH, Matsushita K, Aboyans V, Hess CN, Hicks CW, Kwan TW, et al. 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