猴头菇多糖对氧化应激状态下猪小肠上皮细胞紧密连接相关基因表达的影响

Effects of Hericium erinaceus polysaccharide on gene expression of tight junction in intestinal porcine enterocyte cells induced by oxidative stress

为考察猴头菇多糖(HEP)对氧化应激状态下猪小肠上皮细胞(IPEC-J2)的增殖和紧密连接(TJ)相关基因表达的影响,通过100 g/m L脂多糖(LPS)处理建立IPEC-J2细胞氧化应激模型,在预试验基础上选用125、250、500 g/m L不同质量浓度的HEP处理细胞2、4、8 h,用四甲基偶氮唑盐(MTT)法检测其对正常及氧化应激的IPEC-J2细胞增殖的影响;选取其中最佳的HEP浓度,依上述方法处理细胞,实时荧光定量PCR法检测TJ相关的Occludin、Claudin及ZO-1基因m RNA表达的变化情况。结果显示,不同的HEP浓度和作用时间对正常细胞增殖均有显著影响(P<0.05),对LPS诱导的细胞氧化应激均有缓解作用(P<0.05);HEP的最佳质量浓度为250 g/m L、最佳作用时间为4 h;250 g/m L的HEP作用4、8 h均能显著提高氧化应激状态下,IPEC-J2细胞Occludin、Claudin和ZO-1基因的m RNA表达量(P<0.05)。结果表明,HEP能够促进氧化应激状态下IPEC-J2细胞TJ相关基因的表达,缓解LPS诱导的氧化应激对IPEC-J2屏障功能的损伤。

This study aims to investigate the effects of Hericium erinaceus polysaccharide（HEP） on cell proliferation and gene expression of tight junction in intestinal porcine enterocyte cells （IPEC-J2 cells） induced by oxidative stress. IPEC-J2 cells oxidative stress model was established by LPS（100 μg/mL）. HEP concentration（125,250,500 μg/mL） and treating time（2,4,8 hours） were selected on the basis of preliminary experiment. Cell proliferation was measured by MTT method and expression of tight junction related genes（0ccludin,Claudin,ZO-l） were determined by real-time PCR method. Results showed that HEP with different concentration and treating time could improve normal cell prolifera- tion significantly（P〈0.05） and alleviate the oxidative stress induced by LPS（P〈0.05）. The optimal concen-tration and treating time for HEP were 250 μg/mL and 4 hours,respectively. In addition, HEP（with 250 μg/mL and 4 h or 8 h） could significantly increase the 0ccludin,Claudin and ZO-I gene expression in oxidative-stressedIPEC-J2 cells（P〈0.05）.In conclusion, HEP can promote the expression of tight junc- tion related genes in IPEC-J2 cells to alleviate the injure by oxidative stress in the intestinal epithelial barrier of pigs.