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12周内 这是妇女过早停止母乳喂养的主要原因。人类的机制 乳腺炎大多是未知的，而许多研究已经发表在牛，显示慢性 具有高和延长促炎性物质产生的乳腺组织中的炎性反应 细胞因子（IL-1 B、IL-6、IL-8、TNF-α），已知通过NF-κ B B活性的增加介导。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，因此，产奶量减少， 母乳喂养。有趣的是，许多以前的研究报告了有益的和保护性的 通过激活大麻素2型（CB 2）受体介导的抗炎特性， 内源性大麻素、植物大麻素和合成激动剂在人类慢性 炎症性疾病。此外，发现CB 2表达在这些组织中上调， 慢性炎症反应。先前已经显示用这些激动剂激活CB 2， 导致促炎细胞因子（IL-1B、IL-6、IL-8、TNF-α）的减少，并且ROS的产生也 由NF-κ B B下调介导。事实上，氧化酶（iNOS）的下调和 还观察到抗氧化酶（SOD）的上调。这些是相同的组件， 已知在乳腺炎炎症期间慢性上调的信号通路。总目标 该项目的目的是提供证据证明CB 2在调节中作为治疗靶点的潜在作用。 在乳腺炎感染期间乳腺组织炎症。内源性大麻素和CB 2 在哺乳期的母乳中检测到激动剂2-AG，这提供了该器官可能 在内源性大麻素系统的控制下我们的主要假设是，CB 2表达在 乳腺炎期间乳腺升高，其抗炎特性可通过 内源性和合成的激动剂。这一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CB 2的基因和/或蛋白质表达谱 受体，记录的与CB 2免疫调节相关的分子（NF-κ B、COX 2、iNOS），以及 健康和乳腺炎牛乳腺组织（混合）中的炎性细胞因子（IL-1 β、IL-6、IL-8、TNF-α） 细胞类型），2）测试CB 2受体活化的抗炎作用， 内源性和合成激动剂（2-AG和JWH 133）体外使用牛原代乳腺 通过监测这些上述分子的表达变化进行上皮细胞培养，和3） 表征在S.金黄色葡萄球菌与E.杆菌 监测上述分子表达的变化这些研究预计将 显著推进了目前对CB 2在炎症过程中抗炎潜力理解， 外围器官