谷氨酰胺对高温应激奶牛乳腺上皮细胞凋亡及Bcl-2、HSP70表达的影响  

Effects of Glutamine on Apoptosis and Expression of Bcl-2 and HSP70 in Bovine Mammary Epithelial Cells against Heat Stress

作　　者：张响英[1,2] 唐现文[1] ZHANG Xiangying;TANG Xianwen(Jiangsu Agri-animal Husbandry Vocational College,Taizhou 225300,China;Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]江苏农牧科技职业学院,江苏泰州225300 [2]南京农业大学,江苏南京210095

出　　处：《河南农业科学》2018年第10期111-115,共5页Journal of Henan Agricultural Sciences

基　　金：国家科技支撑计划项目(2012BAD12B10)

摘　　要：为探讨谷氨酰胺预处理抗高温应激损伤的作用,以体外培养的奶牛乳腺上皮细胞为模型,取对数生长期的奶牛乳腺上皮细胞,分为对照组、谷氨酰胺组、高温组、谷氨酰胺+高温组,采用MTT法检测细胞存活率、Annexin V/PI双染法检测细胞凋亡、荧光定量PCR检测Bcl-2基因和HSP70基因的表达水平、比色法分析Caspase-3活性。结果显示,奶牛乳腺上皮细胞经谷氨酰胺预处理后,HSP70 mRNA表达量极显著增加(P <0. 01),提高了热应激状态下细胞的热耐受性和存活力;与高温组相比,谷氨酰胺+高温处理组Bcl-2 mRNA表达量在热恢复0、6、12 h分别提高3. 23倍(P <0. 05)、2. 49倍(P <0. 05)、1. 89倍(P> 0. 05),Caspase-3活性分别下降68. 04%(P <0. 01)、40. 89%(P <0. 05)、52. 06%(P <0. 01)。综上,谷氨酰胺预处理促进了奶牛乳腺上皮细胞Bcl-2和HSP70表达,缓解了高温应激引起的Caspase-3活性上升,抑制了热应激所致的奶牛乳腺上皮细胞凋亡。To explore the effects of glutamine pretreatment on the injury with hyperthermia stress,bovine mammary epithelial cells were cultured in vitro.The logarithmic growth phase cells were divided into control group,glutamine group,hyperthermia group and glutamine+hyperthermia group.Cell survival rate was detected by MTT method,cell apoptosis was measured by Annexin V/PI double staining method,the expression level of Bcl-2 and HSP 70 were tested by FQ-PCR and Caspase-3 activity was detected by colorimetric assay.The results showed that the expression levels of HSP 70 was significantly increased by glutamine pretreatment(P<0.01),the heat tolerance and viability of the mammary cells against heat stress were improved.Compared with hyperthermia group,the expression leves of Bcl-2 mRNA in glutamine+hyperthermia group were 3.23 times(P<0.05),2.49 times(P<0.05)and 1.89 times(P>0.05)in the heat recovery of 0,6,and 12 h respectively,and Caspase-3 activities were decreased by 68.04%(P<0.01),40.89%(P<0.05)and 52.06%(P<0.01)respectively.This suggested that glutamine could enhance the expression levels of Bcl-2 and HSP 70 in bovine mammary epithelial cell,relieve the increase of Caspase-3 activity and inhibit the apoptosis induced by heat stress.

关 键 词：奶牛乳腺上皮细胞 谷氨酰胺 热应激 HSP 70基因 Bcl-2基因 CASPASE-3活性 

分 类 号：S823[农业科学—畜牧学] Q256[农业科学—畜牧兽医]