摘要
为探讨乳酸菌主要代谢产物乳酸抑制肿瘤坏死因子α(TNF-α)诱导的仔猪空肠上皮细胞凋亡的作用以及保护肠黏膜屏障的部分机制,分别使用TNF-α(200μg/L)、乳酸(100μL/L)、TNF-α(200μg/L)+乳酸(100μL/L)处理仔猪空肠上皮细胞(IPEC-J2)组,并设空白对照。用MTT法检测各组的细胞活力;用ELISA方法检测各组caspase-3的蛋白表达量和蛋白活性。结果表明:TNF-α组的细胞存活率极显著低于对照组(P<0.01),乳酸+TNF-α组可显著升高细胞存活率(P<0.05)。乳酸+TNF-α组caspase-3蛋白量和活性显著低于TNF-α组(P<0.05)。乳酸可以抑制TNF-α诱导的IPEC-J2细胞损伤,抑制TNF-α诱导的caspase-3蛋白表达量和蛋白活性的升高,提示乳酸菌主要代谢产物乳酸可通过抑制上皮细胞凋亡来保护肠上皮细胞,进而保护肠黏膜屏障。
The purpose of this study was to determine whether lactic acid, the main product of lactobacillus,could inhibit the apoptosis of IPEC-J2 induced by TNF-alpha, and conduct an initial study on the specificmechanism of lactic acid in protecting the intestinal barrier. Four groups were created： TNF- alpha group,treated with TNF- alpha with the concentration of 200 μg/L; LAC group, treated with lactic acid with theconcentration of 100 μL/L; LAC ＋ TNF- alpha group, treated with TNF- alpha with the concentration of200 μg/L and lactic acid with the concentration of 100 μL/L; control group, treated with maintenance medium.Cell vitality of each group was measured by MTT, proportion of caspase-3 and the vitality of caspase-3 in thetotal protein were detected by ELISA after 12 hours. The results showed that the cell vitality of TNF-alphagroup was significantly lower than that of the control group（P〈0.01）, while cell vitality of LAC＋TNF-alphagroup was significantly higher than that of TNF-alpha group（P〈0.05）. The proportion of caspase-3 and itsvitality in the total protein of the LAC＋TNF-alpha group were lower than those of TNF-alpha group（P〈0.05）.Thus it came to a conclusion that the lactic acid could inhibit the cellular damage induced by TNF-alpha as well as down- regulate the expression and vitality of caspase- 3. The results implied that lactic acid mightprotect the intestinal epithelium through inhibiting the apoptosis of the epithelial cells.
出处
《中国农学通报》
2015年第11期54-58,共5页
Chinese Agricultural Science Bulletin
基金
2014年度国家现代农业产业技术体系"北京生猪产业创新团队岗位专家"(2033213008)
作者简介
第一作者简介：陆相嵩，男，1992年出生，山东聊城人，硕士，研究方向为肠道微生态。通讯地址：102206北京市昌平区回龙观镇北农路7号北京农学院，E-mail：luxiangsong@126．com。
通讯作者：任晓明，男，1958年出生，教授，博士，研究方向为猪病防控与为生态学。通讯地址：102206北京市昌平区回龙观镇北农路7号北京农学院，Tel：010-80795532，E-mail：13910224882@163．com。
