摘要
目的探讨嗜酸乳杆菌代谢产物乳酸(LA)对内毒素(LPS)介导的NF-κB p65入核、转录及环氧化酶-2(COX-2)转录的抑制作用。方法以体外培养的大鼠肠黏膜微血管内皮细胞(RIMMVECs)为材料,设对照组、LPS组、LA预处理组和NF-κB特异性阻断剂吡咯烷二硫代氨基甲酸盐(PDTC)预处理组4个实验组。LPS作用30min后,Western blotting检测不同实验组细胞核、细胞质内NF-κB p65蛋白水平,LPS作用9h后,实时荧光定量PCR法检测NF-κB p65、COX-2 mRNA水平。结果 LPS作用30min后,各实验组细胞NF-κB p65蛋白总量差异不显著,但LA和PDTC预处理组细胞核内NF-κB p65比例显著低于LPS组;LPS作用9h后,LA和PDTC预处理组细胞NF-κB p65和COX-2 mRNA水平显著低于LPS组。结论乳酸具有类似NF-κB阻断剂的作用,可以抑制LPS介导的NF-κB p65入核、转录,抑制NF-κB下游靶基因如COX-2的转录,从而发挥抗炎作用。
Objective To investigate whether lactic acid（LA） ,which was extracted from the culture supernatant of Laetobacillus,could inhibit lipopolysaccharide （LPS）-induced translocation of NF-κB p65 from cytoplasm to nucleus and transcription of NF-κB p65 and cyclooxygenase 2 （ COX-2）. Methods Rat intestinal mucosa microvascular endothelial cells（RIMMVEs） were cultured in vitro and were divided into 4 groups ：control group, LPS group,LA pretreated group and NF-κB inhibitor （pyrrolidinecarbodithioate, PDTC） pretreated group. The protein of NF-κB p65 in cytoplasm and in nucleus was detected by Western blotting analysis after RIMMVEs were treated with LPS for 30 minutes. The mRNA of NF-κB p65 and COX-2 was detected by real time quantity-PCR after RIMMVEs were treated with LPS for 9 hours. Results In a short time（ treated with LPS for 30 minutes） ,the protein expression of NF-κB p65 had no notable difference but the ratios of p65 in nuclei of LA and PDTC pretreated group were notably lower than that of LPS group. After a longer time（treated with LPS for 9 hours） ,the mRNA expression of NF-κB p65 and COX-2 were notably lower than LPS group. Conclusion LA has the effects like NF-KB inhibitor, notably inhibiting LPS-induced translocation of NF-κB p65 from cytoplasm to nucleus and transcription of NF-κB p65 and COX-2. By this way,LA inhibits inflammatory response.
出处
《解剖学报》
CAS
CSCD
北大核心
2013年第2期204-209,共6页
Acta Anatomica Sinica
基金
北京市自然科学基金资助项目(6072007)
北京市教委资助项目(KM200910020002)
