本文選題：乳腺炎 + 乳腺上皮細胞�。� 參考：《西北農(nóng)林科技大學》2017年碩士論文

【摘要】：奶牛乳腺炎是由于乳房損傷或病原微生物入侵所引起的機體代謝和生理狀態(tài)改變的炎癥反應(yīng),是嚴重影響奶牛業(yè)發(fā)展的最常見的疾病之一。在奶牛乳腺發(fā)生炎癥的早期,乳腺導管中的病原菌會首先侵入腺泡表層的乳腺上皮細胞,進而影響乳腺上皮細胞的生理特性,導致上皮細胞的炎性細胞因子異常分泌。乳腺上皮細胞異常分泌的炎性細胞因子通過內(nèi)環(huán)境影響周圍組織中基質(zhì)細胞的生理特性,使基質(zhì)細胞初步表現(xiàn)出炎癥反應(yīng)特征。成纖維細胞是構(gòu)成乳腺組織重要的基質(zhì)細胞,其生長狀態(tài)的改變會直接影響乳腺組織正常的結(jié)構(gòu)和功能,因此研究奶牛乳腺上皮細胞在炎性狀態(tài)下對基質(zhì)成纖維細胞生物特性和功能的功能影響對于奶牛乳腺炎的防治具有重要的意義。金黃色葡萄球菌和大腸桿菌是引起奶牛乳腺炎主要的兩種細菌,脂多糖(lipopolysaccharides,LPS)和脂磷壁酸(lipoteichoic acid,LTA)分別是這兩種導致奶牛乳腺炎的細菌主要毒力成分。本試驗以荷斯坦奶牛乳腺組織分離的原代乳腺上皮細胞作為研究對象,首先利用LPS和LTA對乳腺上皮細胞進行炎癥誘導,確定了LPS和LTA炎癥誘導的最適濃度和最適時間,建立了體外誘導的炎性奶牛乳腺上皮細胞模型;然后通過體外間接共培養(yǎng)模式,以LPS和LTA體外誘導的炎性乳腺上皮細胞與正常基質(zhì)成纖維細胞共培養(yǎng)的方法模擬炎性上皮細胞影響成纖維細胞過程,研究炎性乳腺上皮細胞對基質(zhì)成纖維細胞活性、增殖、遷移、凋亡、炎性細胞因子分泌等生物學特性的影響,同時對導致這些特性發(fā)生變化的分子機制進行探究。結(jié)果表明,LPS和LTA誘導的炎性上皮細胞均可促進了基質(zhì)成纖維細胞的激活,通過MAPK信號通路抑制成纖維細胞的增殖,通過WNT/β-catenin信號通路促進了成纖維細胞的遷移、同時增強成纖維細胞對TNF-α,CCL5,CXCL2,IL6,IL8等炎性相關(guān)因子的表達和分泌,誘導細胞外基質(zhì)蛋白Fibronection和Collagen I的沉積。這些結(jié)果表明奶牛乳腺上皮細胞在炎性狀態(tài)下,通過內(nèi)環(huán)境對基質(zhì)成纖維細胞生炎癥響應(yīng)。這一結(jié)果為進一步研究乳腺基質(zhì)成纖維細胞在奶牛乳腺炎反應(yīng)中的作用及奶牛乳腺炎的防治提供了一定的理論依據(jù)。
[Abstract]:Cow mastitis is an inflammatory response to metabolic and physiological changes caused by breast injury or invasion of pathogenic microorganisms. It is one of the most common diseases that seriously affect the development of dairy cattle industry. In the early stage of breast inflammation in dairy cows, the pathogenic bacteria in mammary duct will first invade the epithelial cells in the surface of the acinus, and then affect the physiological characteristics of the epithelial cells of the mammary gland, leading to the abnormal secretion of inflammatory cytokines in the epithelial cells. The abnormal secretion of inflammatory cytokines by mammary epithelial cells affects the physiological characteristics of stromal cells in the surrounding tissues through the internal environment and makes the stromal cells exhibit the characteristics of inflammatory reaction. Fibroblasts are the most important stromal cells in mammary gland. The change of their growth state will directly affect the normal structure and function of mammary gland. Therefore, it is of great significance to study the effects of dairy mammary epithelial cells on the biological characteristics and function of stromal fibroblasts in inflammatory state for the prevention and treatment of dairy cow mastitis. Staphylococcus aureus and Escherichia coli are two main bacteria causing dairy cow mastitis, lipopolysaccharide polysaccharide LPSs and lipoteichoic acididate LTAs are the main virulence components of these two bacteria. In this experiment, primary mammary epithelial cells isolated from breast tissue of Holstein dairy cattle were used as the research object. First, LPS and LTA were used to induce the inflammation of breast epithelial cells, and the optimal concentration and time of LPS and LTA inflammation induction were determined. The model of inflammatory dairy cow mammary epithelial cells induced in vitro was established, and the model of indirect co-culture in vitro was established. Inflammatory mammary epithelial cells induced by LPS and LTA were co-cultured with normal matrix fibroblasts in vitro to simulate the effect of inflammatory epithelial cells on the process of fibroblasts and to study the activity and proliferation of inflammatory mammary epithelial cells on stromal fibroblasts. The effects of migration, apoptosis, inflammatory cytokine secretion, and the molecular mechanisms that lead to the changes of these characteristics are also explored. The results showed that both lipopolysaccharide and LTA stimulated the activation of stromal fibroblasts, inhibited the proliferation of fibroblasts through MAPK signaling pathway, and promoted the migration of fibroblasts through WNT/ 尾 -catenin signaling pathway. At the same time, the expression and secretion of inflammatory related factors such as TNF- 偽 CCL5, CXCL2, IL6, IL-8 were enhanced in fibroblasts, and the deposition of extracellular matrix proteins (Fibronection and Collagen I) was induced. These results suggest that dairy mammary epithelial cells in inflammatory state, through the internal environment to matrix fibroblast inflammatory response. The results provide a theoretical basis for the further study of the role of mammary stromal fibroblasts in the response of dairy cow mastitis and the prevention and treatment of cow mastitis.
【學位授予單位】：西北農(nóng)林科技大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S858.23

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