转化生长因子β1对奶牛乳腺组织细胞凋亡及HSP70蛋白表达的影响  

Effects of transforming growth factor-β_1 on apoptosis and HSP_(70) expression of mammary tissue of dairy cow

作　　者：狄和双[1] 杨媛媛[1] 王利刚[2] 崔群维[1] 程凯宁[1] 黄静燕[1] 王根林[1] 

机构地区：[1]南京农业大学动物科技学院,江苏南京210095 [2]江苏畜牧兽医职业技术学院,江苏泰州225300

出　　处：《南京农业大学学报》2009年第2期118-123,共6页Journal of Nanjing Agricultural University

基　　金：科技部奶业支撑计划项目(2006BAD04A12,2006BAD04A01);江苏省科技支撑计划项目(BE2008306-2)

摘　　要：应用乳酸脱氢酶（LDH）活力测定、透射电镜、免疫组化和Caspase-3活性分析等技术，检测转化生长因子β1（TGF—β1）对奶牛乳腺组织细胞活性、凋亡和HSP70蛋白表达的影响。用10ng·mL^-1TGF—β1处理乳腺组织48h，分别在不同时间点收集组织和培养液检测各指标，以0h不加TGF—β1作为对照组。结果显示，随TGF—β1作用时间的延长组织细胞活率逐渐降低，24h和48h组显著低于对照组（P〈0．05），LDH活性逐渐升高，6、12、24和48h组极显著高于对照组（P〈0．01）。处理12、24和48h组织DNA发生断裂；Caspase-3活性在24h达到高峰，并极显著高于对照组（P〈0．01）；上皮细胞逐渐由腺泡脱落，腺泡塌陷，但间质结构基本完整；超微结构观察显示，48h时上皮细胞呈现染色质凝集、线粒体肿胀等明显凋亡特征，而成纤维细胞胞浆致密、线粒体丰富，结构较完整、处理3～12h后组织过表达HSP70，且均显著高于对照组（P〈0．05）。结果提示：TGF—β1以促进上皮细胞凋亡的方式促进奶牛乳腺发生退化。The effects of transforming growth factor-β1 （TGF-β1 ） on activity, apoptosis and HSP70 expression of the mammary cells of dairy cow were investigated through multiple methods, including activity test of lactate dehydrogenase （LDH） , electron microscopy analysis, immunohistochemistry analysis and activity analysis of Caspase-3. Mammary tissue was treated with TGF-β1 of 10 ng· mL^-1 for 48 h, then the tissue and its cultures were collected at different time for further analysis. The group without TGF- β1 treatment at 0 h was defined to be control group. The results showed that the survival rate of mammary epithelial cells decreased gradually with the increasing of the time of TGF-β1 treatment, and the survival rates of 24 h and 48 h groups were significantly lower than that of control group （P 〈0.05）. The I,DH activities of 6 h, 12 h, 24 h and 48 h groups were significantly higher than that of control group （ P 〈 0. 01 ） ; the DNA of mammary epithelial eells was degradated in 12 h, 24 h, anti 48 h groups; the activity of Caspase-3 reached to a peak, which was significantly higher than that of control group （ P 〈 0. 01 ）. The acinus of epithelial cells appeared to break off and collapse with the increasing of treatment time, but the leydig structure keeped integrity. Ultrostructure showed that there were obvious apoptosis characteristics in epithelial cells treated for 48 h, such as chromatin condensation and mitochondrial swelling, but the fibroblast cells keeped their compact structure and abundant mitochondrium. HSP70 was overexpressed when treated with TGF-β1 for 3 - 12 h, which were significantly higher than that of control group （ P 〈 0. 05 ）. Conclusion ： TGF-β1 couht induce the apoptosis of the mammary epithelial cells, which might result in the acceleration of mammary involution in dairy cow.

关 键 词：转化生长因子β1(TGF—β1) 乳腺组织 细胞凋亡 HSP70 

分 类 号：S813.3[农业科学—畜牧学]