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作 者:迟世凯 孙春玲[1] 李华涛[1] 丛霞[1] 姜忠玲[1] 王新[1] 曹荣峰[1] 田文儒[1]
机构地区:[1]青岛农业大学动物科技学院,山东青岛266109
出 处:《中国兽医杂志》2015年第6期64-67,共4页Chinese Journal of Veterinary Medicine
基 金:国家自然科学基金(31172378);山东省自然科学基金(ZR2010CM028);山东省农业技术体系创新团队(SDAIT-12-011-03)
摘 要:为了探讨不同浓度黄芩苷对热应激条件下猪肾小管上皮(LLC-PK1)细胞HSP70表达的影响,筛选出黄芩苷诱导热应激条件下细胞HSP70表达的最佳浓度。将培养的LLC-PK1细胞随机分为7个组,Ⅰ组为37℃常温对照组,Ⅱ组为42℃单纯热应激组,Ⅲ、Ⅳ、Ⅴ、Ⅵ组和Ⅶ组分别为42℃热应激并用不同浓度黄芩苷(0.01~100μg/m L)处理组,运用实时荧光定量PCR和Western Blot检测HSP70 m RNA及蛋白的表达。结果显示,42℃单纯热应激能显著诱导LLCPK1细胞HSP70的表达;一定浓度范围内的黄芩苷(0.1~10μg/m L)处理的LLC-PK1细胞HSP70表达显著高于42℃单纯热应激;黄芩苷(0.01~100μg/m L)能显著上调热应激条件下LLC-PK1细胞HSP70的表达,且1μg/m L的黄芩苷上调效果最显著。The objective of the experiment was to explore if the baicalin of different concentrations contributed to HSP70 ex- pression of heat-stressed LLC-PK1 cells and to select the optimal concentration of baicalin for inducing HSPT0 expression. The readily cultured LLC-PKI ceils were divided into seven groups in a random fashion. Cells in group I were served as the control and cultured at 37 ℃ cells in group Ⅱ were heat-stressed at 42 ℃ for 1 h. Group Ⅲ, IV, V, VI and VII were cultured with the addition of different concentrations of baiealin (varying from 0.01 p^g/mL to 100 p^g/mL)at 42 ℃ for 1 h, respectively. The HSP70 mRNA and protein expression were determined by using real-time PCR and western Blot. The experimental results revealed that heat stress at 42 ℃ significantly induce the HSPT0 expression of LLC-PK1 cells. HSP expression of LLC-PK1 cells treated with ba- icalin within certain concentrations (0.1 μg/mL to 10 μg/mL)was significantly elevated than that of the heat-stressed group Ⅱ at 42 ℃.Baicalin(0.01 μg/mL to 100 μg/mL) significantly enhanced HSP70 expression in the cells subjected to heat stress and addi- tion of 1 μg/mL baicalin showed the best effect.
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