摘要
目的探讨不同组合的合生元对CCl_(4)所致急性化学性肝损伤小鼠的肝脏保护作用,及其机制是否与肠—肝轴有关。方法将72只SPF级ICR雄性小鼠随机分12组,每组6只。将含量为5%、15%和20%的麦麸膳食纤维饲料分别记为A1、A2、A3,灌胃剂量为10、20、30 mL/kg的益生菌酸奶分别记为B1、B2、B3,对A1B1组、A1B2组、A1B3组、A2B1组、A2B2组、A2B3组、A3B1组、A3B2组、A3B3组分别进行不同组成合生元干预,阳性对照组、模型组和空白对照组给予普通饲料,阳性对照组同时灌胃复方鳖甲软肝片溶液;30 d后,除空白对照组外的其余各组小鼠灌胃给予1%的CCl_(4)油溶液5 mL/kg,空白对照组灌胃给予等体积玉米油溶液。24 h后,全部小鼠摘眼球取血并分离血清,采用ELISA法检测血清超氧化物歧化酶(SOD)、丙二醛(MDA)水平;取肝脏、结肠、回肠,计算肝脏指数和结肠指数,同时对肝脏和回肠进行HE染色后观察病理改变;收集A1B1组、阳性对照组、模型组和空白对照组小鼠粪便,采用16S rDNA高通量测序法检测肠道菌群情况(Chao、Ace、Shannon、Simpson指数及门水平、属水平上的肠道菌群丰度)。结果模型组肝脏指数及血清MDA水平均高于空白对照组(P均<0.05)。A1B1、A1B2、A2B1和A3B1组小鼠肝脏指数均低于模型组,A1B1、A2B1组小鼠肝脏指数均低于阳性对照组(P均<0.05)。各组小鼠结肠指数比较差异无统计学意义(P>0.05)。A2B1、A2B3组小鼠血清SOD水平均低于模型组、阳性对照组,A1B1、A1B2、A2B3、A3B1和A3B2组小鼠血清MDA水平均低于模型组(P均<0.05);除A1B2组外,其余合生元组小鼠血清MDA水平均高于阳性对照组(P均<0.05)。肝脏病理观察结果:空白对照组小鼠肝组织细胞排列紧密且形态正常,细胞结构清晰且完整,未见炎症细胞浸润和坏死细胞;模型组小鼠肝索、肝小叶结构严重紊乱,细胞肿胀、细胞内存在大量未代谢的CCl_(4);阳性对照组肝细胞形态正常,仅有少量未代谢的CCl_(4);与模型组比较,A1B1组肝小叶结构完整、仅有少量炎症细胞浸润,A2B2组、A3B3组肝小叶严重受损、有大量炎症细胞浸润,A3B1组和A3B3组可见大量未代谢的CCl_(4)。回肠病理观察结果:空白对照组小鼠回肠绒毛较长,绒毛结构完整,肠内皮结构较厚;模型组小鼠回肠结构破坏,肠腔内可见大量黑色物质;阳性对照组绒毛结构较为完整,肠腔内有少量黑色物质;与空白对照组相比,其他合生元组均存在绒毛结构破坏,以A1B1组小鼠绒毛结构完整度最高。模型组小鼠Chao、Ace、Shannon指数均低于空白对照组及A1B1组,Simpson指数高于空白对照组及A1B1组;阳性对照组Shannon指数高于模型组,Simpson指数低于模型组(P均<0.05)。门水平上:与空白对照组比较,模型组小鼠拟杆菌门和变形菌门相对丰度升高,而厚壁菌门相对丰度降低;与模型组比较,A1B1组小鼠厚壁菌门和变形菌门相对丰度均升高,拟杆菌门相对丰度均降低;与阳性对照组比较,A1B1组疣微菌门相对丰度降低(P均<0.05)。属水平上:模型组有10种菌属的相对丰度低于空白对照组,A1B1组7种菌属的相对丰度高于模型组(P均<0.05)。结论合生元对CCl_(4)所致急性化学性肝损伤小鼠具有一定的肝脏保护作用,以5%麦麸膳食纤维联合10 mL/kg益生菌酸奶的合生元组合的肝脏保护效果更好,其机制可能与调节肠—肝轴有关。
Objective To explore the liver protective effect of different combinations of synbiotics on mice with CCl_(4)-induced acute chemical liver injury,and to explore whether the mechanism is related to the gut-liver axis.Methods Seventy-two SPF grade ICR male mice were randomly divided into 12 groups,with six in each.The wheat bran dietary fiber of 5%,15%,and 20%were recorded as A1,A2 and A3,respectively,probiotic yogurt with gastric infusion doses of 10,20,and 30 mL/kg were recorded as B1,B2 and B3,respectively.Mice in the A1B1,A1B2,A1B3,A2B1,A2B2,A2B3,A3B1,A3B2 and A3B3 groups underwent synbiotics intervention of different combinations.Mice in the positive control group,model group and blank control group were given common feed,and mice in the positive control group were simultaneously treated with compound turtle shell soft liver tablet solution.After 30 d,mice of each group except the blank control group were given 5 mL/kg of 1%CCl_(4) oil solution,and mice in the blank control group were given equal volume of corn oil solution.After 24 h,the eyeballs of all mice were removed to collect blood and the serum was isolated.Serum su-peroxide dismutase(SOD)and malondialdehyde(MDA)levels were measured by ELISA.The liver,colon and ileum were taken to calculate the liver index and colon index,and the pathological changes were observed after HE staining of the liver and ileum;the feces of A1B1 group,positive control group,model group and blank control group were collected,and their intestinal flora were analyzed by 16S rDNA high-throughput sequencing(Chao,Ace,Shannon,Simpson indices and abundance of gut microbiota at the phylum and genus levels).Results The liver index and serum MDA level were high-er in the model group than in the blank control group(both P<0.05).The liver indices of mice in the A1B1,A1B2,A2B1 and A3B1 groups were lower than those of the model group,and the liver indices of mice in the A1B1 and A2B1 groups were lower than those of the positive control group(all P<0.05).There was no significant difference in colon index of mice between groups(P>0.05).The serum SOD levels of the A2B1 and A2B3 groups were lower than those of the model and positive control groups,and the serum MDA levels of the A1B1,A1B2,A2B3,A3B1 and A3B2 groups were lower than those of the model group(all P<0.05);except for A1B2 group,the serum MDA levels in the synbiotics groups were higher than those of the positive control group(all P<0.05).Hepatic pathology:mice in the blank control had tightly packed cells with normal morphology,the cell structure was clear and complete,and no inflammatory cell infiltration and necrotic cells were observed;mice in the model group had severe disturbances of the hepatic cord and lobular structure of the liver with cell swelling and the presence of abundant unmetabolized CCl_(4) in the cells;mice in the positive control group had a normal hepatocyte morphology with only a few unmetabolized CCl_(4);compared with the model group,A1B1 group had in-tact hepatic lobule structure with only few inflammatory cells,A2B2 and A3B3 groups had severely damaged liver lobules and massive infiltration of inflammatory cells,and large amounts of unmetabolized CCl_(4) were seen in the A3B1 and A3B3 groups.Pathological observations of the ileum:intestinal villus in mice of the blank control group were long,with com-plete villus structure and thick intestinal endothelial structure;in the model group,a large amount of black material could be seen in the intestinal lumen;intestinal villus in the positive control group was complete with a small amount of black ma-terial in the intestinal lumen;compared with the blank control group,all other synbiotics groups had villus structure de-struction,with the highest integrity of villus structure in the A1B1 group.Chao,Ace,Shannon index were lower in the model group than in the blank control group and A1B1 group,Simpson index was higher than those of the blank control group and A1B1 group;Shannon index was higher in the positive control group than in the model group,and Simpson in-dex was lower than that of the model group(all P<0.05).At phylum level:compared with the blank control group,the rel-ative abundances of Bacteroidetes and Proteobacteria increased in the model group,while the relative abundance of Fir-micutes decreased;in the A1B1 group,the relative abundance of Bacteroidetes decreased;in the positive control group,the relative abundance of A1B1 group decreased(all P<0.05).At the genus level:the relative abundances of 10 species in the model group were lower than those of the blank control group,and the relative abundances of 7 species in the A1B1 group were higher than those of the model group(all P<0.05).Conclusion Synosbiotics have liver protective effect on mice with acute chemical liver injury.The liver protective effect of combination of 5%wheat bran dietary fiber and 10 mL/kg probiotic yogurt is better,and the mechanism may be related to the regulation of gut-liver axis.
作者
李艳艳
丁红
刘承尧
郭宇沨
LI Yanyan;DING Hong;LIU Chengyao;GUO Yufeng(School of Public Health,Xinjiang Medical University,Urumqi 830011,China)
出处
《山东医药》
CAS
2023年第11期37-42,共6页
Shandong Medical Journal
基金
新疆医科大学研究生科研创新项目(CXCY2022036)。
