不同泌乳相关激素和生长因子对奶牛乳腺上皮细胞增殖的影响

摘要

本研究旨在探讨不同泌乳相关激素和生长因子对奶牛乳腺上皮细胞增殖的影响及其与细胞外基质主要成分层黏连蛋白的关系.将正常的荷斯坦泌乳期奶牛乳腺上皮细胞进行体外培养,在未包被或包被层黏连蛋白的条件下,以MTT法检测催乳素(PRL)、牛生长激素(GH)、类胰岛素生长因子-1(IGF-Ⅰ)、类胰岛素生长因子-2(IGF-Ⅱ)对细胞增殖作用的影响.在层黏连蛋白包被条件下,进行血清恢复的同时添加不同泌乳相关激素和生长因子,GH、IGF-Ⅰ有促进细胞增殖的作用(P＜0.05),PRL、IGF-Ⅱ有维持细胞存活的作用(P＜0.05)；无血清时,几种激素和生长因子单独添加均无明显促增殖效应(P＞0.05).无基质条件下,与血清联合使用时,PRL、GH、IGF-Ⅰ、IGF-Ⅱ均对细胞生长有不同程度促进作用(P＜0.05);无血清时,仅IGF-Ⅰ使细胞增殖速率显著加快(P＜0.05).层黏连蛋白作为培养基质对于体外培养的泌乳乳腺上皮细胞生长速度没有显著促进作用,但有利于PRL和IGF-Ⅱ发挥促存活作用.PRL、GH、IGF-I、IGF-Ⅱ对细胞增殖和存活有促进作用,但需要与血清中其他成分协同才能充分发挥作用,其中IGF-Ⅰ促增殖能力最强.%To explore the relationship between the influence of proliferation and laminin belong to extracellular matrix in mammary epithelial cell of dairy cow from different hormone and cytokine related lactation. Took the normal mammary glands tissue in lactation dairy cow and cultured the mammary epithelial cells in vitro, deteminate the influence of prolactin (PRL), bovine growth hormone (GH), insulin-like growth factor-1 (IGF- I ) , insulin-like growth factor-2 (IGF- Ⅱ) on the proliferation of mammary epithelial cells in dairy cow with MTT under laminin coating or not. The facilitation in proliferation with blood serum, differernt hormone and cytokine related lactation was found by GH and IGF- I (P<0. 05) under laminin coating, cell survival was maintained by PRL and IGF-Ⅱ (P<0. 05). Under the condition of un-blood serum, different hormone and cytokine related lactation haplo-addition has phanero-less proliferation (P>0. 05). Cell growth has different encouraged by PRL, GH, IGF- I and IGF-Ⅱ with laminin uncoating and blood serum (P<0. 05); the proliferation was higher up-setted by IGF-Ⅰ (P<0. 05). Laminin as culture matrix is encourage-less to the normal mammary epithelial cells in lactation dairy cow cultured in vitro, which is beneficial for auxo-survival by PRL and IGF- Ⅱ. The action of proliferation and survival were encouraged by PRL, GH, IGF-Ⅰ and IGF -Ⅱ , which needed blood serum. The proliferation ability of IGF- I is highest.