雌激素对奶牛乳腺上皮细胞增殖及抗氧化性的影响  

Effects of Estrogen on Proliferation and Antioxidation in Bovine(Bos taurus) Mammary Epithelial Cells

作　　者：陈静[1] 王根林[1] 李莲[1] 于静[1] 赵士杰[1] 蒋小强[1] 王斐[1] 

机构地区：[1]南京农业大学动物科技学院,南京210095

出　　处：《农业生物技术学报》2013年第2期216-222,共7页Journal of Agricultural Biotechnology

基　　金："十二五"国家科技计划课题(No.2011BAD28B02);江苏省农业三新工程课题(No.SX2011-189)

摘　　要：雌激素在哺乳动物生理中期过程发挥重要作用。本研究旨在探讨不同浓度的雌二醇(estradiol,E2)对体外培养奶牛(Bos taurus)乳腺上皮细胞增殖、抗氧化性的影响。实验分溶剂对照组(CK)和0.5、1、5、10、100nmol/LE2处理组(Ⅰ～Ⅴ),培养奶牛乳腺上皮细胞4、8和12h后,MTT法检测细胞增殖情况;在培养12h后,比色法检测胞外乳酸脱氢酶(LDH)、胞内超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量;荧光定量RT-PCR法检测细胞中HSP70(热休克蛋白70)mRNA的表达量。结果表明:(1)与CK相比,Ⅰ～Ⅲ组细胞存活率升高(P<0.05);Ⅳ、Ⅴ组细胞存活率降低。(2)Ⅰ～Ⅳ组胞外LDH活性降低,胞内HSP70mRNA表达量、SOD活性高于对照组(P<0.05),脂质化反应减弱;Ⅴ组胞外LDH活性升高(P<0.05),细胞的HSP70mRNA表达量、SOD活性低于对照组(P<0.05),脂质化反应增强。本研究结论:低浓度(0.5、1.0和5.0nmol/L)E2能够促进奶牛乳腺上皮细胞的增殖,诱导乳腺上皮细胞中HSP70mRNA表达,进而通过调节超氧化物歧化酶(SOD)活性发挥抗氧化作用,长期处于高浓度(100nmol/L)E2环境下,对乳腺细胞增殖及抗氧化性则起抑制作用并引起细胞损伤。本研究为应用外源性雌激素提高牛乳腺组织抗氧化性提供了一定理论依据。Estrogen plays an important role for mammalian in interim physiology period. The objective of this study was to reserch the effects of estradiol （E2） on cells proliferation and antioxidation in vitro bovine（Bos taurus） mammary epithelial cells （BMEC）. Experimental groups were divided into I - V, which were treated with E2 at 0.5, 1, 5, 10 and 100 nmol/L, respectively, the control group （CK） received the vehicle only. After culturing for 4, 8 and 12 h, the viability of mammary epithelial cells was assessed by MTT assay. The 12 h culture medium and cells were used to determin the content of malonaldehyde （MDA）, the activities of Lactate dehydrogenase （LDH） and superoxide dismutase （SOD） by colorimetric method. HSP70（heat shock protein 70） mRNA expression were assayed by Real-time fluorescent quantitative PCR. The results showed： （1） Compared with the control group, I -Ⅲ E2-treated groups showed markedly increased cell viabilities （P〈0.05）. IV and V E2-treated groups decreased cell viabilities. （2） I -IV E2-treated groups showed a decrease in LDH activity in culture medium, HSP70 mRNA expression and SOD activity were significantly higher than those in the control group（P〈0.05） but lipid reaction was reduced. V Ez-treated group showed a significant increase in LDH activity in culture medium （P〈0.05）, a decrease of HSP70 mRNA expression and SOD activity （P〈0.05） but lipid reaction was enhanced. The conclusion of this study： Low concentrations （0.5, 1.0 and 5.0 nmol/L） Ez could promote the proliferation of mammary epithelial cells, induced expression of HSP70 mRNA, which might play antioxidant role by increasing intracellular SOD activity. High concentration （100 nmol/L） of E2 could attenuate the proliferation and antioxidation of mammary epithelial cells, causing cell damage with long-term stimulation. This study provided some theoretical basis that exogenous estrogens can increase the antioxidant activity of bovine mammary tissue.

关 键 词：雌二醇 奶牛乳腺上皮细胞 增殖 抗氧化 热休克蛋白70(HSP70) 

分 类 号：S827.1[农业科学—畜牧学]