山豆根多糖对IPEC-J2细胞氧化损伤的保护作用  

Protective effects of Sophora subprostrate polysaccharide against oxidative dam-age in IPEC-J2 cells

作　　者：许双 杨琨兆 郭鑫 陈怡沁 严思茵 何政科 苏利娟 马琪 董世起 曹立亭[1,2] 杜红旭 XU Shuang;YANG Kunzhao;GUO Xin;CHEN Yiqin;YAN Siyin;HE Zhengke;SU Lijuan;MA Qi;DONG Shiqi;CAO Liting;DU Hongxu(College of Veterinary Medicine,Southwest University,Rongchang,Chongqing 402460,China;Institute of Traditional Chinese Veterinary Medicine,Southwest University,Rongchang,Chongqing 402460,China)

机构地区：[1]西南大学动物医学院,重庆荣昌402460 [2]西南大学中兽医药研究所,重庆荣昌402460

出　　处：《中国兽医学报》2025年第2期330-340,共11页Chinese Journal of Veterinary Science

基　　金：重庆市自然科学基金资助项目(CSTB2022NSCQ-MSX0470);西南大学博士基金资助项目(SWU120002);国家生猪技术创新中心先导科技资助项目(NCTIP-XD/B12);重庆市现代农业产业技术体系创新团队资助项目(CQMAITS202312)。

摘　　要：旨在探究山豆根多糖(Sophora subprostrate polysaccharide, SSP)对叔丁基过氧化氢(tert-butyl hydroperoxide, TBHP)诱导的猪小肠上皮细胞(IPEC-J2)氧化损伤的保护作用及可能机制。通过MTT法确定TBHP的最佳造模剂量和SSP的安全浓度范围。试验分为5组:空白组(CC组)、模型组(TBHP组)、SSPL组、SSPM组和SSPH组,观察细胞形态并测定细胞存活率和LDH释放量,利用DCFH-DA染色检测细胞内ROS的含量,通过生化检测试剂盒检测细胞中的MDA的含量以及抗氧化指标的变化,利用转录组技术分析SSP缓解IPEC-J2细胞氧化损伤的可能机制。结果显示,625μmol/L TBHP作用2 h,可以显著降低IPEC-J2细胞活性,显著提高LDH的释放量(P<0.05),显著抑制CAT、SOD和GPX活性(P<0.05),明显提高MDA和ROS的含量(P<0.05)。与模型组相比,SSP处理后,细胞内ROS的含量显著降低(P<0.05),同时CAT、SOD和GPX活力显著地提高(P<0.05),MDA含量和LDH释放量显著降低(P<0.05),且呈剂量依赖性。转录组分析发现,TBHP处理明显改变了IPEC-J2细胞的转录谱,而SSP处理则可以一定程度恢复受损细胞的转录谱变化,同时GO与KEGG富集分析表明,CC与TBHP组之间差异表达基因主要显著富集在氧化磷酸化、核糖体等通路上。而SSP与TBHP组之间的差异表达基因主要富集在氧化磷酸化、凋亡与乙醛酸和二羧酸酯代谢等通路。结果表明,TBHP可能是通过氧化磷酸化和影响甘氨酸,丝氨酸和苏氨酸代谢等代谢通路,干扰IPEC-J2细胞正常氧化呼吸链从而引起肠上皮细胞氧化受损。而SSP处理可能通过作用于氧化磷酸化、凋亡和溶酶体等信号通路,恢复肠上皮细胞氧化磷酸化过程,缓解溶酶体损伤,减少细胞的凋亡来减轻肠上皮细胞氧化损伤。这一发现为SSP在缓解猪肠道氧化损伤的临床应用中提供了理论依据。The objective of this study was to evaluate the protective effect and possible related mechanisms of Sophora subprostrate polysaccharide(SSP)on intestinal epithelial cell injury in-duced by Tert-Butyl hydroperoxide(TBHP).The optimal dose of TBHP and the safe concentra-tion range of SSP were determined using the MTT method.In this study,IPEC-J2 cells were divid-ed into five groups:the control group,the model group,the SSPL group,the SSPM group and the SSPH group,and the cell morphology,cell survival rate and LDH release rate were observed and measured.The content of intracellular reactive ROS was observed and determined by DCFH-DA staining.The content of MDA in the supernatant and the antioxidant index of cells were determined by the reagent kit.Transcriptome technology was employed to analyze the potential mechanisms by which SSP mitigates oxidative damage in IPEC-J2 cells.The results showed that treatment with 625μmol/L TBHP for 2 h significantly reduced the activity of IPEC-J2 cells,markedly increased LDH release(P<0.05),inhibited CAT superoxide SOD and glutathione GPX activities(P<0.05),and significantly elevated MDA and ROS levels(P<0.05).Compared to the model group,after SSP treatment,intracellular ROS levels were significantly reduced(P<0.05),while CAT,SOD,and GPX activities were significantly increased(P<0.05),and MDA content and LDH re-lease were significantly decreased(P<0.05)in a dose-dependent manner.Transcriptome analysis revealed that TBHP treatment significantly altered the transcriptional profiles of IPEC-J2 cells,while SSP treatment could restore the transcriptional profiles of the damaged cells to a certain ex-tent.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)indicated that the differentially expressed genes between the CC and TBHP groups were significantly enriched in oxidative phosphorylation,ribosome,and other pathways.Meanwhile,the differentially expressed genes between the SSP and TBHP groups were mainly enriched in oxidative phosphorylation,ap-optosis,glyoxylate and di

关 键 词：山豆根多糖 叔丁基过氧化氢 IPEC-J2细胞 氧化应激 

分 类 号：S831.5[农业科学—畜牧学]