酿酒酵母培养物对内毒素诱导的鹅肠上皮细胞炎性损伤的缓解作用  

Alleviating Effects of Yeast Culture on Remission of Inflammatory Injury Induced by Lipopolysaccharide in Goose Intestinal Epithelial Cells

作　　者：闫乐艳[1] 邵春荣[1] 李悦 闫俊书[1] 奚雨萌 YAN Leyan;SHAO Chunrong;LI Yue;YAN Junshu;XI Yumeng(Key Laboratory of Crop and Livestock Integrated Farming,Ministry of Agriculture,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)

机构地区：[1]江苏省农业科学院畜牧研究所农村农业部种养结合重点实验室,南京210014

出　　处：《动物营养学报》2022年第11期7392-7401,共10页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金项目(31902190);江苏省农业科技自主创新资金项目[CX(21)2013]。

摘　　要：本试验旨在研究酿酒酵母培养物(YC)对内毒素(LPS)诱导的原代鹅肠上皮细胞炎性损伤的缓解作用。试验利用混合酶消化法成功获得原代鹅肠上皮细胞,通过1μmol/L LPS处理建立肠上皮细胞炎性损伤模型;并分别选取浓度为100、300和500 mg/L的YC溶液预处理细胞,分析YC对LPS诱发的炎性损伤的缓解作用。试验采用CCK-8法测定细胞活性,实时荧光定量PCR检测炎性因子及核转录因子-κB(NF-κB)信号通路基因表达,荧光探针法分析细胞活性氧(ROS)浓度,并通过测定抗氧化酶活性评价细胞氧化应激状态。结果表明:1μmol/L LPS可诱导肠上皮细胞白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和白细胞介素-8(IL-8)基因相对表达量显著上调(P<0.05),同时显著或极显著上调Toll样受体2A(TLR2A)(P<0.01)、Toll样受体4(TLR4)(P<0.01)、髓样分化因子88(MyD88)(P<0.05)、NF-κB(P<0.01)等NF-κB信号通路基因相对表达量;而使用300和500 mg/L浓度的YC预处理可抑制LPS诱导的炎症反应,减少ROS生成。同时,300 mg/L的YC预处理还可显著缓解LPS引起的细胞超氧化物歧化酶(SOD)活性下降(P<0.05),降低细胞丙二醛(MDA)和一氧化氮(NO)含量(P<0.05)。由此可见,YC可调节鹅肠上皮免疫平衡,缓解炎性损伤,还可通过提高肠上皮细胞对氧自由基的清除能力及抗氧化能力,保护肠上皮细胞的正常功能。The present study aimed to demonstrate the alleviating effects of yeast culture(YC)on inflammatory injury induced by lipopolysaccharide(LPS)in goose intestinal epithelial cells.The primary intestinal epithelial cells of goose were cultured in vitro and established the inflammatory injury model of intestinal epithelial cells with LPS.Then,the cells were pretreated with YC at concentrations of 100,300 and 500 mg/L to analyze the alleviating effects of YC on LPS induced inflammatory injury.The cell viability was determined by CCK-8,the gene expressions of cytokines and the nuclear factor-κB(NF-κB)signaling pathway was detected by RT-PCR,the concentration of reactive oxygen species(ROS)was determined by fluorescence probe techniques,and the oxidative stress status was evaluated by measuring antioxidant enzyme activities.The results showed as follows:LPS upregulated the relative expression levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),interleukin-8(IL-8)(P<0.05),Toll-like receptor 2A(TLR2A)(P<0.01),toll-like receptor 4(TLR4)(P<0.01),myeloid differentiation factor 88(MyD88)(P<0.05)and NF-κB(P<0.01)gens in intestinal epithelial cells.Then,the YC pretreatment at the 300 and 500 mg/L levels could inhibit LPS induced inflammatory response and oxidative stress,reducing the ROS production and maintaining the cellular functions.The YC pretreatment at the 300 mg/L level also increased the activity of superoxide dismutase(SOD)(P < 0.05)which was effected by LPS,and also decreased and the contents of malondialdehyde(MDA)(P<0.05)and nitric oxide(NO)(P<0.05).In conclusion,these results suggest that YC can regulate the immune balance of goose intestinal epithelial cells,inhibit inflammatory reactions,and improve the scavenging ability of oxygen free radicals,protecting intestinal epithelial cells from inflammatory injury and oxidative stress.[Chinese Journal of Animal Nutrition,2022,34(11):7392-7401]

关 键 词：酿酒酵母培养物 内毒素 鹅肠上皮细胞 炎性损伤 抗氧化功能 

分 类 号：S835[农业科学—畜牧学]