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The response surface methodology (RSM) based on the Box\u2013Behnken design (BBD) was used to optimize the deproteinization conditions as follows: TCA: n\u2010butanol\u00a0=\u00a01: 5.1, polysaccharide solution: (TCA\u2013n\u2010butanol)\u00a0=\u00a01: 2.8, and shook for 33\u2009min. LJP could promote the thymus and spleen indexes of cyclophosphamide (CTX)\u2010induced immune\u2010deficient mice. Besides, the contents of cytokine interleukin\u20102 (IL\u20102) and hemolysin in mice serum were augmented after treatment with LJP. Based on serum metabolomics analysis, a total of 14 metabolites (VIP\u2009&gt;1.0, FC\u2009&gt;2 or FC\u2009&lt;0.5, and <jats:italic>p<\/jats:italic> value\u2009&lt;\u2009.05) were selected as the potential biological biomarkers related to the LJP for treating CTX\u2010induced mice. After the pathway enrichment analysis, these metabolites were mainly involved in the relevant pathways of arginine biosynthesis, Citrate cycle, and other metabolic pathways. Network pharmacology further showed that there were 57 key targeting proteins in the intersection of the potential biological biomarkers and immunodeficiency\u2010related targeting proteins according to protein\u2013protein interactions analysis (PPI). The biological function analysis indicated that the potential biological processes were mainly associated with tricarboxylic acid (TCA) cycle, phospholipid metabolic process, metabolic process, and so on. In conclusion, serum metabolomics combined with network pharmacology could be helpful to clarify the immunomodulatory mechanism of LJP and provide a literature basis for further clinical research on the therapeutic mechanism of LJP.<\/jats:p>","DOI":"10.1002\/fsn3.3067","type":"journal-article","created":{"date-parts":[[2022,9,25]],"date-time":"2022-09-25T18:06:15Z","timestamp":1664129175000},"page":"364-378","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Optimization of protein removal process of <i>Lonicera japonica<\/i> polysaccharide and its immunomodulatory mechanism in cyclophosphamide\u2010induced mice by metabolomics and network pharmacology"],"prefix":"10.1002","volume":"11","author":[{"given":"Jie","family":"Ding","sequence":"first","affiliation":[{"name":"School of Pharmaceutical Sciences Shandong University of Traditional Chinese Medicine  Jinan 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