为研究无乳链球菌对奶牛乳腺成纤维细胞(BMFB)瘦素(Leptin)及其长型受体(OB-Rb)表达的影响,本研究将不同浓度热灭活的无乳链球菌(0、105 c fu/mL、106 cfu/mL和108 cfu/mL)作用于BMFB,分别于不同作用时间采用荧光定量PCR (qPCR)方法检测Leptin及OB-Rb mRNA转录水平,western blot方法检测Leptin及OB-Rb蛋白的表达.各组总体变化规律结果显示,无乳链球菌作用BMFB后,除6h、12 h 106 cfu/mL和108 cfu/mL作用浓度外,其余作用时间实验组Leptin mRNA的转录水平及蛋白表达量与对照组相比显著升高(p＜0.05);各作用时间实验组OB-Rb mRNA的转录水平及蛋白表达量与对照组相比极显著升高(p＜0.01).上述研究结果表明,热灭活的无乳链球菌能够促进BMFB的Leptin及OB-Rb mRNA的转录水平和蛋白的表达,为奶牛乳腺纤维化的防治提供依据.%In order to explore the effect of Streptococcus agalactiae on the expression of Leptin and OB-Rb in bovine mammary fibroblasts (BMFB),the BMFB cultured in vitro were exposed to the heat-inactivated S.agalactiae at 105 cfu/mL,106 cfu/mL,108 cfu/mL,respectively.The total RNA and protein were extracted from the cells in different time (6 h,12 h,24 h,48 h,72 h),and the mRNA transcriptional level and the protein expression of leptin and OB-Rb in BMFB were detected by qPCR and western blot.The results showed that the mRNA transcriptional level and protein expressions of Leptin in BMFB treated by S.agalactiae in experimented group were higher than that in control group (p-0.05) except for that in BMFB treated with 106 and 108 cfu/mL in 6 h and 12 h with 106 cfu/mL and 108 cfu/mL,the OB-Rb mRNA transcriptional level and protein expressions in expermental group were significantly higher than that in control group (p＜0.01) at each time.The above results indicated that heat-inactivated S.agalactiae could promote the mRNA transtription and the expression of Leptin and OB-Rb in BMFB,which provided the basic for preventwn and trentment of manimarg gland fibrosis in dainy cow.
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