Investigating the therapeutic potential of CB2 in modulation of inflammation in the mammary gland during lactation

研究 CB2 在调节哺乳期乳腺炎症方面的治疗潜力

• 批准号: 10514546
• 负责人: Juliana Perez Laspiur
• 金额: $ 28.27万
• 依托单位: UNIVERSITY OF PUERTO RICO AT AGUADILLA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10514546
• 关键词: 2-arachidonylglycerol; Agonist; Alzheimer' s Disease; Anti-Bacterial Agents; Anti-Inflammatory Agents; Antiinflammatory Effect; Antioxidants; Atherosclerosis; Bacterial Infections; Breast Diseases; Breast Epithelial Cells; Breast Feeding; CNR2 gene; Cannabinoids; Cannabis; Cattle; Cell Culture Techniques; Chronic; Collaborations; Country; Cyclic AMP; Disease; Down-Regulation; Endocannabinoids; Enzymes; Escherichia coli; Faculty; Gene Expression; Goals; Human; Human Milk; IL8 gene; Immune response; In Vitro; Infant Malnutrition; Infant Mortality; Infection; Inflammation; Inflammatory; Inflammatory Response; Innate Immune Response; Interleukin-1; Interleukin-6; Intestines; Knowledge; Lactation; Mammary gland; Mediating; Messenger RNA; Modeling; Monitor; Mothers; Multiple Sclerosis; NF-kappa B; Nerve Degeneration; Organ; Oxidants; PTGS2 gene; Peptides; Peripheral; Phagocytes; Plants; Postpartum Period; Process; Production; Property; Proteins; Publishing; Reactive Oxygen Species; Receptor Activation; Reporting; Rheumatoid Arthritis; Role; Signal Pathway; Signal Transduction; Staphylococcus aureus; Staphylococcus aureus infection; Symptoms; Syndrome; TNF gene; Testing; Therapeutic; Tissues; Up-Regulation; Woman; antioxidant enzyme; base; cell type; chronic inflammatory disease; cost effective treatment; cytokine; endogenous cannabinoid system; human model; immunoregulation; mammary; mastitis; milk production; phytocannabinoid; prevent; protein expression; recruit; synthetic cannabinoid; therapeutic target; transcription factor; treatment strategy

Mastitis is an inflammatory breast disease that develops mostly during the first 12 weeks’ post-partum in women and it is the major reason for early cessation of breastfeeding. The mechanisms of human mastitis are mostly unknown, while many studies have been published in bovine, showing a chronic inflammatory response in mammary tissue with high and extended production of pro-inflammatory cytokines (IL-1B, IL-6, IL-8, TNF-), known to be mediated via increases in the activity of NF-B. During chronic inflammation these cytokines, reactive oxygen species (ROS), and phagocytes can cause considerable damage to the mammary tissue and hence, decrease in milk production and cessation of breastfeeding. Interestingly, numerous previous studies have reported the beneficial and protective anti-inflammatory properties mediated by the activation of the cannabinoid type 2 (CB2) receptor with endogenous cannabinoids, phytocannabinoids, and synthetic agonists in human models of chronic inflammatory diseases. Furthermore, CB2 expression is found upregulated in these tissues undergoing chronic inflammatory responses. Activation of CB2 with these agonists has been previously shown to causes a decrease in pro-inflammatory cytokines (IL-1B, IL-6, IL-8, TNF-) and ROS production also mediated by a downregulation of NF-B. Indeed, downregulation of oxidative enzymes (iNOS) and upregulation of anti-oxidant enzymes (SOD) is also observed. These are the same components and signaling pathways known to be chronically upregulated during mastitis inflammation. The overall goal of this project is to provide evidence of the potential role of CB2 as a therapeutic target in the modulation of mammary tissue inflammation during mastitis infection. The endogenous cannabinoid and CB2 agonist 2-AG has been detected in human milk during lactation, providing evidence this organ may be under control of the endocannabinoid system. Our main hypothesis is that CB2 expression in the mammary gland is elevated during mastitis, and its anti-inflammatory properties can be activated by endogenous and synthetic agonists in vitro. This hypothesis will be tested through three specific aims in bovine mammary tissue: 1) to characterize the gene and/or protein expression profile of the CB2 receptor, documented molecules related to CB2 immunomodulation (NF-K, COX2, iNOS), and inflammatory cytokines (IL-1, IL-6, IL-8, TNF-) in healthy and mastitis bovine mammary tissue (mixed cell types) during lactation, 2) to test the anti-inflammatory effect of CB2 receptor activation via endogenous and synthetic agonists (2-AG and JWH133) in vitro using bovine primary mammary epithelial cell culture by monitoring changes in expression of these aforementioned molecules, and 3) characterize host response to CB2 activation during infection with S. aureus compared to E. Coli by monitoring changes in expression of these aforementioned molecules These studies are expected to significantly advance current understanding of CB2 anti-inflammatory potential during inflammation of peripheral organs.

乳腺炎是一种炎症性乳腺疾病，主要发生在产后前 12 周内 对于女性来说，这是提前停止母乳喂养的主要原因。人体的机制 乳腺炎大多是未知的，但许多研究已发表在牛身上，显示出慢性 乳腺组织的炎症反应，促炎物质大量且长期产生 细胞因子（IL-1B、IL-6、IL-8、TNF-α），已知通过 NF-β 活性增加来介导。期间 这些细胞因子、活性氧 (ROS) 和吞噬细胞可引起慢性炎症 对乳腺组织造成相当大的损害，从而导致产奶量减少和停止 哺乳。有趣的是，许多先前的研究报告了有益和保护性的 通过激活 2 型大麻素 (CB2) 受体介导的抗炎特性 慢性病人类模型中的内源性大麻素、植物大麻素和合成激动剂 炎症性疾病。此外，发现这些组织中的 CB2 表达上调 慢性炎症反应。先前已证明用这些激动剂激活 CB2 导致促炎细胞因子（IL-1B、IL-6、IL-8、TNF-α）和 ROS 产生减少 由 NF-κB 下调介导。事实上，氧化酶（iNOS）和 还观察到抗氧化酶（SOD）的上调。这些是相同的组件并且 已知在乳腺炎炎症期间长期上调的信号通路。总体目标 该项目的目的是提供 CB2 作为调节治疗靶点的潜在作用的证据 乳腺炎感染期间乳腺组织炎症。内源性大麻素和 CB2 哺乳期间在人乳中检测到激动剂 2-AG，这提供了该器官可能是 在内源性大麻素系统的控制下。我们的主要假设是 CB2 表达于 乳腺炎期间乳腺升高，其抗炎特性可以通过 体外内源性和合成激动剂。该假设将通过三个具体目标进行检验 在牛乳腺组织中：1) 表征 CB2 的基因和/或蛋白质表达谱 受体，已记录的与 CB2 免疫调节相关的分子（NF-K、COX2、iNOS），以及 健康牛乳腺组织和乳腺炎牛乳腺组织中的炎性细胞因子（IL-1、IL-6、IL-8、TNF-）（混合） 细胞类型）在哺乳期间，2）通过测试 CB2 受体激活的抗炎作用 使用牛初级乳腺进行体外内源性和合成激动剂（2-AG 和 JWH133） 通过监测上述分子表达的变化来进行上皮细胞培养，以及3) 与大肠杆菌相比，表征金黄色葡萄球菌感染期间宿主对 CB2 激活的反应 监测上述分子表达的变化这些研究预计将 显着推进了目前对 CB2 在炎症过程中抗炎潜力的理解 周围器官。