D-天冬氨酸对溃疡性结肠炎小鼠肠道细胞坏死关键信号分子及肠道微生物组成的影响  

Effects of D⁃Aspartate on Key Signaling Molecules for Intestinal Cell Necrosis and Intestinal Microbiota Composition of Ulcerative Colitis Mice

作　　者：黄乐 何羿文 刘思危 王晨昱 陈铭哲 范高升 冯舒锾 印遇龙[1,2,3] 李铁军 何流琴[1,3] HUANG Le;HE Yiwen;LIU Siwei;WANG Chenyu;CHEN Mingzhe;FAN Gaosheng;FENG Shuhuan;YIN Yulong;LI Tiejun;HE Liuqin(Hunan Provincial Key Laboratory of Animal Nutritional Physiology and Metabolic Process,National Engineering Laboratory for Pollution Control and Waste Utilization in Livestock and Poultry Production,Institute of Subtropical Agriculture,Chinese Academy of Sciences,Changsha 410125,China;College of Advanced Agricultural Sciences,University of Chinese Academy of Science,Beijing 100049,China;Hunan Provincial Key Laboratory of Animal Intestinal Function and Regulation,College of Life Sciences,Hunan Normal University,Changsha 410081,China)

机构地区：[1]中国科学院亚热带农业生态研究所,畜禽养殖污染控制与资源化技术国家工程实验室,动物营养生理与代谢过程湖南省重点实验室,长沙410125 [2]中国科学院大学现代农业科学学院,北京100049 [3]湖南师范大学生命科学学院,动物肠道功能与调控湖南省重点实验室,长沙410081

出　　处：《动物营养学报》2024年第10期6710-6722,共13页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：广西省自然科学面上项目(2022JJA130333);国家自然科学基金(32172755,3210099);国家自然科学区域创新联合基金重点支持项目(U23A20233);湖南省科技创新领军人才项目(2023RC1054);湖南省重点研发项目(2022NK2023)。

摘　　要：本试验旨在探讨D-天冬氨酸(D-Asp)对葡聚糖硫酸钠(DSS)诱导的溃疡性结肠炎(UC)小鼠肠道细胞坏死关键信号分子及肠道微生物组成的影响。试验选取40只C57BL/6J雄性小鼠,初始体重为(20.98±0.14)g,随机分为5组:对照组(CON组)、葡聚糖硫酸钠组(DSS组)、低浓度D-天冬氨酸组(L-Dasp组)、中浓度D-天冬氨酸组(M-Dasp组)、高浓度D-天冬氨酸组(H-Dasp组)。每组8个重复,每个重复1只小鼠。试验前7 d,CON组小鼠饮用蒸馏水,其他各组小鼠饮用含3.5%DSS的蒸馏水;从第8天开始,CON组和DSS组小鼠直肠灌注100μL生理盐水,L-Dasp组、M-Dasp组和H-Dasp组直肠灌注100μL浓度分别为0.15、2.30和7.70 mmol/L的D-Asp溶液,每隔1 d灌注1次,共灌注5次。结果表明:1)与CON组相比,DSS组结肠细胞凋亡率显著升高(P<0.05),线粒体出现明显肿胀,呈空泡状,且数量减少(P>0.05)。与DSS组相比,L-Dasp、M-Dasp和H-Dasp组的结肠细胞凋亡率均显著降低(P<0.05),结肠细胞线粒体数量均显著升高(P<0.05)。2)与CON组相比,DSS组的结肠Toll样受体4(TLR4)和白细胞介素-1β(IL-1β)mRNA相对表达量显著升高(P<0.05);与DSS组相比,M-Dasp组的结肠TLR4 mRNA相对表达量显著降低(P<0.05)。各组之间结肠白细胞介素-1α(IL-1α)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)含量无显著差异(P>0.05)。3)与CON组相比,DSS组的结肠受体相互作用蛋白激酶1(RIPK1)蛋白相对表达量显著升高(P<0.05);与DSS组相比,L-Dasp、M-Dasp和H-Dasp组的结肠RIPK1蛋白相对表达量显著降低(P<0.05),H-Dasp组的结肠受体相互作用蛋白激酶3(RIPK3)蛋白相对表达量显著降低(P<0.05)。4)与CON组相比,DSS组的结肠放线菌门(Actinobacteria)相对丰度显著升高(P<0.05);与DSS组相比,L-Dasp组、M-Dasp组和H-Dasp组的结肠放线菌门相对丰度显著降低(P<0.05)。与CON组相比,DSS组的结肠另枝菌属(Alistipes)相对丰度显著降低(P<0.05),结肠芽孢杆菌属(Bacillus)和糖多孢菌This study aimed to investigate the effects of D-aspartate(D-Asp)on the key signal molecules for intestinal cell necrosis and intestinal microbiota composition of ulcerative colitis(UC)mice induced by dextran sodium sulfate(DSS).Forty C57BL/6J male mice with similar body weight of(20.98±0.14)g were ran-domly selected and divided into 5 groups:control group(CON group),dextran sodium sulfate group(DSS group),low concentration D-Asp group(L-Dasp group),medium concentration D-Asp group(M-Dasp group)and high concentration D-Asp group(H-Dasp group).Each group included 8 replicates with 1 mouse in each replicate.Seven days before the experiment,mice in the CON group were given distilled water,while mice in other groups were given distilled water containing 3.5%DSS.From day 8,mice in CON and DSS groups were rectal perfused with 100μL of saline,while mice in L-Dasp,M-Dasp and H-Dasp groups were rectal perfused with solution containing 0.15,2.30 and 7.70 mmol/L D-Asp,respectively,perfused once ev-ery other day,and 5 times of perfusion were performed.The results showed as follows:1)compared with the CON group,the apoptosis rate of colonic cells of DSS group was significantly increased(P<0.05),the mito-chondria were observed swelling,vacuolated,and the number was reduced(P>0.05).Compared with the DSS group,the apoptosis rate of colonic cells of L-Dasp,M-Dasp and H-Dasp groups was significantly de-creased(P<0.05),and the number of mitochondria in colon cells was significantly increased(P<0.05).2)Compared with the CON group,the mRNA relative expression levels of Toll-like receptor 4(TLR4)and inter-leukin-1β(IL-1β)in colon of DSS group were significantly increased(P<0.05);compared with the DSS group,the mRNA relative expression level of TLR4 in colon of M-Dasp group was significantly decreased(P<0.05).There were no significant differences in contents of nterleukin-1α(IL-1α),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in colon among all groups(P>0.05).3)Compared with the CON group,the protein relative expression

关 键 词：D-天冬氨酸 溃疡性结肠炎 细胞凋亡 肠道微生物 受体相互作用蛋白信号通路 

分 类 号：S811[农业科学—畜牧学]