摘要
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.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2017年第1期128-134,共7页
Chinese Veterinary Science
基金
国家自然科学基金项目(31502110)
关键词
猴头菇多糖
猪小肠上皮细胞
氧化应激
紧密连接
Hericium erinaceus polysaccharide
intestinal porcine enterocyte cells
oxidative stress
tight junction