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This study focuses on the role of miR-125b-5p in modulating apoptosis in HCT-8 cells in response to\n                      <jats:italic>C<\/jats:italic>\n                      .\n                      <jats:italic>parvum<\/jats:italic>\n                      infection.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Expression levels of miR-125b-5p and\n                      <jats:italic>C<\/jats:italic>\n                      .\n                      <jats:italic>parvum<\/jats:italic>\n                      burden were estimated using real-time quantitative polymerase chain reaction (RT-qPCR) and cell apoptosis was detected by flow cytometry. The interaction between miR-125b-5p and Bak1 was studied by luciferase reporter assay, RT-qPCR, and western blotting. The expression of Cytochrome c (Cyt c), Caspase-3, and Caspase-9 were assessed by RT-qPCR, and western blotting. JC-1 assess mitochondrial membrane potential.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      MiR-125b-5p expression decreased at 3, 6, 12, and 24\u00a0h post-infection (hpi) but increased at 48 and 72\u00a0hpi. Upregulation of miR-125b-5p inhibited apoptosis and increased parasite burden in HCT-8 cells following\n                      <jats:italic>C<\/jats:italic>\n                      .\n                      <jats:italic>parvum<\/jats:italic>\n                      infection, whereas its downregulation promoted apoptosis and reduced the parasite burden. Luciferase reporter assays identified Bak1 as a target gene of miR-125b-5p. Suppression of Bak1 expression via small interfering RNA (siRNA) inhibited apoptosis and increased the\n                      <jats:italic>C<\/jats:italic>\n                      .\n                      <jats:italic>parvum<\/jats:italic>\n                      burden, while Bak1 overexpression promoted apoptosis and reduced the parasite burden. Co-transfection with miR-125b-5p mimics and pcDNA3.1-Bak1 demonstrated that miR-125b-5p regulates apoptosis and parasite burden by targeting Bak1. Further analysis showed that overexpression of miR-125b-5p significantly decreased the mRNA and protein levels of Cytochrome c (cyt-c), caspase-3, and caspase-9, as well as reduced mitochondrial membrane damage. Inhibition of miR-125b-5p produced opposite effects. Co-transfection with miR-125b-5p mimics and pcDNA3.1-Bak1 revealed that pcDNA3.1-Bak1 counteracted the inhibitory effects of miR-125b-5p on cyt-c, caspase-3, caspase-9, and mitochondrial membrane damage.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      The present data indicated that miR-125b-5p targets Bak1 to regulate apoptosis in HCT-8 cells in response to\n                      <jats:italic>C<\/jats:italic>\n                      .\n                      <jats:italic>parvum<\/jats:italic>\n                      infection via the mitochondrial pathway.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Graphical Abstract<\/jats:title>\n                  <\/jats:sec>","DOI":"10.1186\/s13071-025-07033-1","type":"journal-article","created":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T13:48:45Z","timestamp":1761659325000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["MiR-125b-5p targets Bak1 to regulate HCT-8 cell apoptosis in response to Cryptosporidium parvum infection via mitochondrial pathway"],"prefix":"10.1186","volume":"18","author":[{"given":"Shanbo","family":"Wu","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tianren","family":"Shao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juanfeng","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jingjing","family":"Xie","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lulu","family":"Sun","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yafang","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lijie","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoying","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Junqiang","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yayun","family":"Wu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qingda","family":"Meng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Longxian","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rongjun","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,10,28]]},"reference":[{"key":"7033_CR1","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1016\/S0140-6736(13)60844-2","volume":"382","author":"KL Kotloff","year":"2013","unstructured":"Kotloff KL, et al. 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