{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,26]],"date-time":"2025-12-26T05:20:56Z","timestamp":1766726456692,"version":"3.48.0"},"reference-count":66,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,12,24]],"date-time":"2025-12-24T00:00:00Z","timestamp":1766534400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the National Center of Pratacultural Technology Innovation (under preparation) Special Fund for Innovation Platform Construction","award":["CCPTZX2023B06"],"award-info":[{"award-number":["CCPTZX2023B06"]}]},{"name":"the Central Government Guided Local Science and Technology Development Fund Project","award":["2024ZY0115"],"award-info":[{"award-number":["2024ZY0115"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>This experiment was designed to systematically evaluate the immunomodulatory effect of water extract of Artemisia annua L. (WEAA) on sheep, both in vivo and in vitro, and to determine the involvement of the TLR4\/NF-\u03baB signaling pathway in mediating these effects. In experiment 1, 32 female sheep (Dorper \u00d7 Han, 3 months old, 24 \u00b1 0.09 kg each) were designated to 4 groups, with each group receiving a basal diet supplemented with, respectively, 0 (control group), 500, 1000, and 1500 mg\/kg WEAA. The serum, liver, and spleen immune indicators and related gene expressions were measured. In experiment 2, the peripheral blood lymphocytes (PBLs) were processed with WEAA (0, 25, 50, 100, 200, and 400 \u03bcg\/mL), with six replicates assigned to each concentration group, then the cell viability, immune function, and related gene expressions were measured, and the optimal concentration of WEAA was determined. In experiment 3, the experimental groups consisted of PBLs subjected to treatments with or without PDTC (NF-\u03baB inhibitor) and with or without WEAA, forming four distinct treatment groups (six replicates\/group): PDTC(\u2212)\/WEAA(\u2212) group, PDTC(\u2212)\/WEAA(+) group, PDTC(+)\/WEAA(\u2212) group and PDTC(+)\/WEAA(+) group. The immune indexes and TLR4\/NF\u03baB pathway related indexes were determined. The results were as follows: WEAA dose-dependently enhanced the content of immunoglobulins (IgA, IgG, IgM) and cytokines (IL-1\u03b2, IL-2, IL-4) in the serum, liver, and spleen tissues, among which IgA, IgG, and IL-4 were the most significantly affected core indicators (p &lt; 0.05). Meanwhile, WEAA dose-dependently upregulated the expression of TLR4\/NF-\u03baB pathway-related genes (TLR4, IKK\u03b2, I\u03baB\u03b1, NF-\u03baBp65) and their downstream cytokine-related genes (IL-1\u03b2, IL-4) in liver and spleen tissues (p &lt; 0.05). Of these genes, liver IL-4, I\u03baB\u03b1, and spleen IL-4 were the most prominently regulated core genes (p &lt; 0.05), The optimal supplementary dose of WEAA was determined to be 1000 mg\/kg. In addition, adding 100 \u03bcg\/mL WEAA to the culture medium of PBLs significantly enhanced immune function and cell viability. The underlying mechanism involved the TLR4\/NF-\u03baB pathway; that is to say, WEAA enhanced sheep\u2019s immune indicators by upregulating TLR4\/NF-\u03baB pathway genes, thereby coordinately regulating humoral and innate immunity, thereby improving the immune indices of sheep. This study provided compelling experimental support for the prospective utilization of WEAA as a functional feed supplement in intensive meat-type sheep production systems.<\/jats:p>","DOI":"10.3390\/ani16010059","type":"journal-article","created":{"date-parts":[[2025,12,24]],"date-time":"2025-12-24T16:41:19Z","timestamp":1766594479000},"page":"59","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Immunomodulatory Effect of Artemisia annua L. Water Extract on Meat-Type Sheep via Activating TLR4\/NF-\u03baB Signaling Pathways"],"prefix":"10.3390","volume":"16","author":[{"given":"Gen","family":"Gang","sequence":"first","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruiheng","family":"Gao","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shiwei","family":"Guo","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu","family":"Xin","sequence":"additional","affiliation":[{"name":"National Center of Pratacultural Technology Innovation (Under Preparation), Hohhot 010060, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4623-3566","authenticated-orcid":false,"given":"Xiao","family":"Jin","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuanyuan","family":"Xing","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sumei","family":"Yan","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuanqing","family":"Xu","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8550-3509","authenticated-orcid":false,"given":"Binlin","family":"Shi","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Ding, W., Lu, Y., Xu, B., Chen, P., Li, A., Jian, F., Yu, G., and Huang, S. 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