{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,23]],"date-time":"2025-12-23T15:38:08Z","timestamp":1766504288524,"version":"build-2065373602"},"reference-count":32,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:00:00Z","timestamp":1760140800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005847","name":"Health and Medical Research Fund","doi-asserted-by":"crossref","award":["#15161391","#16172691"],"award-info":[{"award-number":["#15161391","#16172691"]}],"id":[{"id":"10.13039\/501100005847","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Research Grants Council of Hong Kong GRF","award":["PolyU15100821M"],"award-info":[{"award-number":["PolyU15100821M"]}]},{"name":"NFSC\/RGC schemes","award":["N_PolyU 520\/20"],"award-info":[{"award-number":["N_PolyU 520\/20"]}]},{"name":"Hong Kong Polytechnic University Project of Strategic Importance","award":["ZE2C"],"award-info":[{"award-number":["ZE2C"]}]},{"name":"RISA"},{"name":"RiFood seed funds","award":["CD71","CD54"],"award-info":[{"award-number":["CD71","CD54"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Background\/Objectives: Osteoporosis is a prevalent and debilitating skeletal disease characterized by a progressive loss of bone mass and deterioration of bone microarchitecture. Probiotics have emerged as a potential therapeutic tool for treating osteoporosis through modulation of the gut microbiota. In this study, we aimed to examine the effects of live Lacticaseibacillus rhamnosus AC1 (LR-AC1), isolated from a fecal sample from a newborn in Hong Kong, on deoxycorticosterone acetate (DOCA)-induced bone loss in a rat model. Methods: Bone mass and microarchitecture were assessed using micro-computed tomography (micro-CT). Immunostaining for CD31+ and osterix, markers of endothelial cells and osteoblast precursors, respectively, was performed. Gut microbiota composition was analyzed via 16S rRNA sequencing. The effects of an LR-AC1 cell-free conditioned supernatant (CCS) on osteoclastogenesis, angiogenesis, and migration of bone marrow mesenchymal stem cells (BMSCs) were evaluated in vitro using RT-qPCR and wound healing assays. Results: LR-AC1 administration did not induce adverse effects in healthy rats; however, it exacerbated bone loss in rats with DOCA-salt-induced osteoporosis. Correspondingly, the number of CD31-positive endothelial cells and osterix-positive osteoprogenitors decreased with bone loss. In vitro, LR-AC1 CCS promoted osteoclastogenesis and angiogenesis, while in the presence of DOCA, LR-AC1 CCS inhibited BMSC migration. Gut microbiota analysis revealed that the relative abundances of the genera g_RF39 and g_Clostridia_UCG-014 correlated with the severity of bone loss. Conclusions: While several studies suggest that probiotics can prevent and treat osteoporosis, our findings indicate that in a male rat model of DOCA-salt-induced osteoporosis, live LR-AC1 aggravated bone loss. This effect is associated with alterations in gut microbiota and disruption of the coupling process in bone remodeling.<\/jats:p>","DOI":"10.3390\/nu17203198","type":"journal-article","created":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T07:51:28Z","timestamp":1760428288000},"page":"3198","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Lacticaseibacillus rhamnosus AC1 Aggravates Bone Loss in a Male Rat Model of Deoxycorticosterone Acetate (DOCA)-Salt-Induced Osteoporosis"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6491-5398","authenticated-orcid":false,"given":"Xiaoqing","family":"Kuang","sequence":"first","affiliation":[{"name":"Department of Food Science and Nutrition, Faculty of Science, The Hong Kong Polytechnic University, Hung Hom, Hong Kong SAR, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haicui","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Food Science and Nutrition, Faculty of Science, The Hong Kong Polytechnic University, Hung Hom, Hong Kong SAR, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4149-2983","authenticated-orcid":false,"given":"Tim Fat","family":"Shum","sequence":"additional","affiliation":[{"name":"Department of Food Science and Nutrition, Faculty of Science, The Hong Kong Polytechnic University, Hung Hom, Hong Kong SAR, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1949-7822","authenticated-orcid":false,"given":"Chunyi","family":"Wen","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Faculty of Engineering, The Hong Kong Polytechnic University, Hung Hom, Hong Kong SAR, China"},{"name":"Research Institute of Smart Ageing, The Hong Kong Polytechnic University, Hung Hom, Hong Kong SAR, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8306-1826","authenticated-orcid":false,"given":"Jiachi","family":"Chiou","sequence":"additional","affiliation":[{"name":"Department of Food Science and Nutrition, Faculty of Science, The Hong Kong Polytechnic University, Hung Hom, Hong Kong SAR, China"},{"name":"Research Institute for Future Food, The Hong Kong Polytechnic University, Hung Hom, Hong Kong SAR, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,11]]},"reference":[{"key":"ref_1","first-page":"111","article-title":"Vertebral compression fractures in the elderly","volume":"69","author":"Old","year":"2004","journal-title":"Am. Fam. Physician"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1357","DOI":"10.1002\/jbmr.1588","article-title":"The gut microbiota regulates bone mass in mice","volume":"27","author":"Sjogren","year":"2012","journal-title":"J. Bone Miner. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1038\/nature10213","article-title":"Human nutrition, the gut microbiome and the immune system","volume":"474","author":"Kau","year":"2011","journal-title":"Nature"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1016\/j.disamonth.2015.03.011","article-title":"Probiotics","volume":"61","author":"Mizock","year":"2015","journal-title":"Dis. Mon."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1080\/07315724.2017.1318724","article-title":"Effects of a Multispecies Probiotic Supplement on Bone Health in Osteopenic Postmenopausal Women: A Randomized, Double-blind, Controlled Trial","volume":"36","author":"Jafarnejad","year":"2017","journal-title":"J. Am. Coll. Nutr."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3169","DOI":"10.1210\/EN.2015-1308","article-title":"Loss of Bone and Wnt10b Expression in Male Type 1 Diabetic Mice Is Blocked by the Probiotic Lactobacillus reuteri","volume":"156","author":"Zhang","year":"2015","journal-title":"Endocrinology"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1793","DOI":"10.1002\/jcp.24340","article-title":"Probiotic use decreases intestinal inflammation and increases bone density in healthy male but not female mice","volume":"228","author":"McCabe","year":"2013","journal-title":"J. Cell Physiol."},{"key":"ref_8","unstructured":"Yasir, M., Goyal, A., and Sonthalia, S. (2023). Corticosteroid Adverse Effects. StatPearls, StatPearls Publishing."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1038\/s41440-021-00796-x","article-title":"Hypotensive effect of captopril on deoxycorticosterone acetate-salt-induced hypertensive rat is associated with gut microbiota alteration","volume":"45","author":"Wu","year":"2022","journal-title":"Hypertens. Res."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Chan, L.C., Zhang, Y., Kuang, X., Koohi-Moghadam, M., Wu, H., Lam, T.Y.C., Chiou, J., and Wen, C. (2022). Captopril Alleviates Chondrocyte Senescence in DOCA-Salt Hypertensive Rats Associated with Gut Microbiome Alteration. Cells, 11.","DOI":"10.3390\/cells11193173"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Wu, H., Shum, T.F., and Chiou, J. (2021). Characterization of the Probiotic Potential of Lactic Acid Bacteria Isolated from Kimchi, Yogurt, and Baby Feces in Hong Kong and Their Performance in Soymilk Fermentation. Microorganisms, 9.","DOI":"10.3390\/microorganisms9122544"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"792","DOI":"10.1002\/ptr.1739","article-title":"Concurrent administration of aqueous Azadirachta indica (neem) leaf extract with DOCA-salt prevents the development of hypertension and accompanying electrocardiogram changes in the rat","volume":"19","author":"Obiefuna","year":"2005","journal-title":"Phytother. Res."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Wu, H.C., Jiang, L.L., Shum, T.F., and Chiou, J. (2022). Elucidation of Anti-Hypertensive Mechanism by a Novel AC1 Fermented Soymilk in the Deoxycorticosterone Acetate-Salt Hypertensive Rats. Nutrients, 14.","DOI":"10.3390\/nu14153174"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2867","DOI":"10.1007\/s10620-019-05628-0","article-title":"GG Ameliorates Liver Injury and Hypoxic Hepatitis in Rat Model of CLP-Induced Sepsis","volume":"64","author":"Ding","year":"2019","journal-title":"Digest Dis. Sci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"100227","DOI":"10.1016\/j.bonr.2019.100227","article-title":"Characterizing how probiotic Lactobacillus reuteri 6475 and lactobacillic acid mediate suppression of osteoclast differentiation","volume":"11","author":"Quach","year":"2019","journal-title":"Bone Rep."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Schulter, A., Weller, P., Kanaan, O., Nel, I., Heusgen, L., Hoing, B., Hasskamp, P., Zander, S., Mandapathil, M., and Dominas, N. (2018). CD31 and VEGF are prognostic biomarkers in early-stage, but not in late-stage, laryngeal squamous cell carcinoma. BMC Cancer, 18.","DOI":"10.1186\/s12885-018-4180-5"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1124","DOI":"10.1158\/0008-5472.CAN-04-2128","article-title":"Osterix, a transcription factor for osteoblast differentiation, mediates antitumor activity in murine osteosarcoma","volume":"65","author":"Cao","year":"2005","journal-title":"Cancer Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1002\/jbmr.3947","article-title":"Involvement of the Gut Microbiota and Barrier Function in Glucocorticoid-Induced Osteoporosis","volume":"35","author":"Schepper","year":"2020","journal-title":"J. Bone Miner. Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/S1043-2760(00)00234-4","article-title":"Glucocorticoid-induced osteoporosis","volume":"11","author":"Manelli","year":"2000","journal-title":"Trends Endocrinol. Metab."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Sapra, L., Dar, H.Y., Bhardwaj, A., Pandey, A., Kumari, S., Azam, Z., Upmanyu, V., Anwar, A., Shukla, P., and Mishra, P.K. (2021). Lactobacillus rhamnosus attenuates bone loss and maintains bone health by skewing Treg-Th17 cell balance in Ovx mice. Sci. Rep., 11.","DOI":"10.1038\/s41598-020-80536-2"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2049","DOI":"10.1172\/JCI86062","article-title":"Sex steroid deficiency-associated bone loss is microbiota dependent and prevented by probiotics","volume":"126","author":"Li","year":"2016","journal-title":"J. Clin. Investig."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2040622319860653","DOI":"10.1177\/2040622319860653","article-title":"Lactobacillus rhamnosus GG attenuates tenofovir disoproxil fumarate-induced bone loss in male mice via gut-microbiota-dependent anti-inflammation","volume":"10","author":"Liu","year":"2019","journal-title":"Ther. Adv. Chronic Dis."},{"key":"ref_23","first-page":"E591","article-title":"Mild stimulatory effect of a probiotic mix on bone mass when treatment is initiated 1.5 weeks after ovariectomy in mice","volume":"320","author":"Ohlsson","year":"2021","journal-title":"Am. J. Physiol. Metab."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"104762","DOI":"10.1016\/j.jff.2021.104762","article-title":"Anti-osteoporotic potential of Lactobacillus plantarum AR237 and AR495 in ovariectomized mice","volume":"87","author":"Yu","year":"2021","journal-title":"J. Funct. Foods"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Di Maggio, N., and Banfi, A. (2022). The osteo-angiogenic signaling crosstalk for bone regeneration: Harmony out of complexity. Curr. Opin. Biotech., 76.","DOI":"10.1016\/j.copbio.2022.102750"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"426","DOI":"10.7150\/thno.34126","article-title":"Type H blood vessels in bone modeling and remodeling","volume":"10","author":"Peng","year":"2020","journal-title":"Theranostics"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"9793","DOI":"10.1039\/D1FO01468D","article-title":"Gut microbiota-dependent catabolites of tryptophan play a predominant role in the protective effects of turmeric polysaccharides against DSS-induced ulcerative colitis","volume":"12","author":"Yang","year":"2021","journal-title":"Food Funct."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Roth, W., Zadeh, K., Vekariya, R., Ge, Y., and Mohamadzadeh, M. (2021). Tryptophan Metabolism and Gut-Brain Homeostasis. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22062973"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"110778","DOI":"10.1016\/j.exger.2019.110778","article-title":"The multiple faces of tryptophan in bone biology","volume":"129","author":"Sharma","year":"2020","journal-title":"Exp. Gerontol."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Anaya, J.M., Bollag, W.B., Hamrick, M.W., and Isales, C.M. (2020). The Role of Tryptophan Metabolites in Musculoskeletal Stem Cell Aging. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21186670"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1083\/jcb.201006123","article-title":"The two faces of serotonin in bone biology","volume":"191","author":"Ducy","year":"2010","journal-title":"J. Cell Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1007\/s00223-022-00975-z","article-title":"Lactobacillus rhamnosus GG Promotes Intestinal Vitamin D Absorption by Upregulating Vitamin D Transporters in Senile Osteoporosis","volume":"111","author":"Cheng","year":"2022","journal-title":"Calcif. Tissue Int."}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/17\/20\/3198\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T04:24:49Z","timestamp":1760502289000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/17\/20\/3198"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,11]]},"references-count":32,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2025,10]]}},"alternative-id":["nu17203198"],"URL":"https:\/\/doi.org\/10.3390\/nu17203198","relation":{},"ISSN":["2072-6643"],"issn-type":[{"type":"electronic","value":"2072-6643"}],"subject":[],"published":{"date-parts":[[2025,10,11]]}}}