Gratz S, Wu Q K, El-Nezami H, Juvonen R O, Mykkänen H, Turner P C
Department of Clinical Nutrition, University of Kuopio, P.O. Box 1627, 70211 Kuopio, Finland.
Appl Environ Microbiol. 2007 Jun;73(12):3958-64. doi: 10.1128/AEM.02944-06. Epub 2007 Apr 20.
The probiotic Lactobacillus rhamnosus GG is able to bind the potent hepatocarcinogen aflatoxin B1 (AFB1) and thus potentially restrict its rapid absorption from the intestine. In this study we investigated the potential of GG to reduce AFB1 availability in vitro in Caco-2 cells adapted to express cytochrome P-450 (CYP) 3A4, such that both transport and toxicity could be assessed. Caco-2 cells were grown as confluent monolayers on transmembrane filters for 21 days prior to all studies. AFB1 levels in culture medium were measured by high-performance liquid chromatography. In CYP 3A4-induced monolayers, AFB1 transport from the apical to the basolateral chamber was reduced from 11.1%+/-1.9% to 6.4%+/-2.5% (P=0.019) and to 3.3%+/-1.8% (P=0.002) within the first hour in monolayers coincubated with GG (1x10(10) and 5x10(10) CFU/ml, respectively). GG (1x10(10) and 5x10(10) CFU/ml) bound 40.1%+/-8.3% and 61.0%+/-6.0% of added AFB1 after 1 h, respectively. AFB1 caused significant reductions of 30.1% (P=0.01), 49.4% (P=0.004), and 64.4% (P<0.001) in transepithelial resistance after 24, 48, and 72 h, respectively. Coincubation with 1x10(10) CFU/ml GG after 24 h protected against AFB1-induced reductions in transepithelial resistance at both 24 h (P=0.002) and 48 h (P=0.04). DNA fragmentation was apparent in cells treated only with AFB1 cells but not in cells coincubated with either 1x10(10) or 5x10(10) CFU/ml GG. GG reduced AFB1 uptake and protected against both membrane and DNA damage in the Caco-2 model. These data are suggestive of a beneficial role of GG against dietary exposure to aflatoxin.
益生菌鼠李糖乳杆菌GG能够结合强效肝癌致癌物黄曲霉毒素B1(AFB1),从而有可能限制其从肠道的快速吸收。在本研究中,我们调查了GG在体外对适应表达细胞色素P-450(CYP)3A4的Caco-2细胞中AFB1可用性的影响,以便同时评估转运和毒性。在所有研究之前,Caco-2细胞在跨膜滤器上生长为汇合单层21天。通过高效液相色谱法测量培养基中的AFB1水平。在CYP 3A4诱导的单层中,与GG(分别为1x10(10)和5x10(10) CFU/ml)共同孵育的单层中,AFB1从顶侧室到基底外侧室的转运在第一小时内从11.1%±1.9%降至6.4%±2.5%(P = 0.019),并降至3.3%±1.8%(P = 0.002)。1小时后,GG(1x10(10)和5x10(10) CFU/ml)分别结合了40.1%±8.3%和61.0%±6.0%的添加AFB1。AFB1分别在24、48和72小时后导致跨上皮电阻显著降低30.1%(P = 0.01)、49.4%(P = 0.004)和64.4%(P < 0.001)。在24小时后与1x10(10) CFU/ml GG共同孵育可在24小时(P = 0.002)和48小时(P = 0.04)时防止AFB1诱导的跨上皮电阻降低。仅用AFB1处理的细胞中可见DNA片段化,但与1x10(10)或5x10(10) CFU/ml GG共同孵育的细胞中未见。GG在Caco-2模型中减少了AFB1的摄取,并防止了膜和DNA损伤。这些数据表明GG对饮食中黄曲霉毒素暴露具有有益作用。