摘要
研究无乳链球菌(GBS)对奶牛乳腺成纤维细胞(BMFB)转化生长因子β_1(TGF-β_1)及其受体TβRⅠ表达的影响。6代和7代BMFB,在6、12、24、48h时间点,分别用无血清培养液和10~5、10~6、10~8CFU/mL的不同浓度的热灭活GBS作用于BMFB细胞。RT-qPCR法检测TGF-β_1及TβRⅠmRNA的表达,Western blot法检测TGF-β_1和TβRⅠ蛋白的表达。与对照组相比,TGF-β_1mRNA的表达量明显升高,于24h达到最大值(P<0.05);TGF-β_1蛋白的表达于12h达到最大值,24h蛋白表达量开始明显降低(P<0.05)。TβRⅠmRNA的表达量较对照组的升高,在48h达到峰值(P<0.05);TβRⅠ蛋白表达量在12h低于6h,24h达到峰值,之后又下降(P<0.05)。说明热灭活无乳链球菌对BMFB TGF-β_1及TβRⅠmRNA和蛋白的表达有促进作用。
In order to investigate the effects of different concentrations of heat-inactivated Streptococcus agalactiae on the expressions of transforming growth factor β1 （TGF-β1） and its receptor TβR I in bovine mammary fibroblasts （BMFB）, 6 or 7 generations of cultured in vitro BMFBs were exposed to the serumfree medium and the heat-inactivated GBS at different concentrations （10 5, 10 6, 10 8CFU/mL）,respectively,and the total RNA and proteins were extracted from the cells after 6,12,24 and 48 hour, respectively. The mRNA expressions of TGF-β1 and TβR I were detected by real-time fluorescence quantitative PCR, The protein expressions of TGF-β1 and TβR I were detected by Western-blot.Compared with the control group,the mRNA expression of TGF-β1 was significantly elevated when BMFB were exposed to GBS, and reached the maximal levels at 24 h（P〈0.05）.The protein expression of TGF-β1 reached the maximal levels at 12 h,and then the expression went down at 24 h（P〈0.05）.The expression of TβR I mRNA was higher than that of the control group,and reached the maximal levels at 48 h （P〈0.05）.The expression of TβR I protein at 12 h was lower than that at 6 h,and reached the maximal levels at 48 h,and then the expression went down （P 〈0.05）. The preliminary study confirmed that heat-inactivated Streptococcus agalactiae could promote the expressions of TGF-β1 and TβR I mRNA and protein in BMFBs.
出处
《动物医学进展》
北大核心
2017年第8期7-13,共7页
Progress In Veterinary Medicine
基金
国家自然科学基金项目(31460642)
