橙皮苷和迷迭香酸组合对脂多糖攻毒大鼠回肠形态、菌群结构以及炎症反应的影响

Effects of Hesperidin and Rosmarinic Acid Combination on Ileal Morphology,Microbiota Structure and Inflammatory Response of Lipopolysaccharide-Challenged Rats

本试验旨在探究橙皮苷和迷迭香酸组合对脂多糖(LPS)攻毒大鼠回肠形态、菌群结构以及炎症反应的影响。将32只21日龄的无特定病原体(SPF)级SD大鼠随机分为4组,分别为LPS组、Hes组、RA组、Hes×RA组,每组4个重复,每个重复2只大鼠(公母各占1/2)。各组大鼠均饲喂标准基础饲粮,Hes组大鼠每日灌胃300 mg/kg BW的橙皮苷,RA组大鼠每日灌胃20 mg/kg BW的迷迭香酸,Hes×RA组每日灌胃150 mg/kg BW橙皮苷+10 mg/kg BW迷迭香酸,LPS组每日灌胃等量的生理盐水。灌胃处理共进行14 d,在第13天,所有大鼠腹腔注射剂量为500μg/kg BW的LPS溶液,用于诱导急性肠道损伤模型。结果显示:与LPS组相比,Hes×RA组大鼠回肠绒毛高度和绒隐比显著提高(P<0.05)。高通量测序结果显示,与LPS组相比,Hes×RA组大鼠回肠食糜微生物的丰富度和多样性均没有显著变化(P>0.05);Hes×RA组大鼠回肠食糜中乳杆菌属(Lactobacillus)的相对丰度显著提高(P<0.05),放线菌门(Actinobacteria)、变形菌门(Proteobacteria)、棒状杆菌属(Corynebacterium)和链球菌属(Streptococcus)的相对丰度显著降低(P<0.05);Hes×RA组大鼠回肠食糜微生物代谢产物乙酸、丙酸和总短链脂肪酸的含量显著升高(P<0.05)。与LPS组相比,Hes×RA组大鼠回肠黏膜中髓过氧化物酶(MPO)活性以及白细胞介素-6(IL-6)含量显著降低(P<0.05),白细胞介素-10(IL-10)含量显著提高(P<0.05)。与LPS组相比,Hes×RA组大鼠回肠黏膜中封闭蛋白-1(Claudin-1)和闭锁蛋白(Occludin)的mRNA相对表达量显著提高(P<0.05),闭锁小带蛋白-1(ZO-1)的mRNA相对表达量有增加的趋势(P=0.064)。与LPS组相比,Hes×RA组大鼠回肠黏膜中白细胞介素-1受体相关激酶1(IRAK1)和肿瘤坏死因子受体相关蛋白6(TRAF6)的mRNA相对表达量显著降低(P<0.05)。综上可知,橙皮苷和迷迭香酸组合可以改善LPS攻毒引起的大鼠回肠形态损伤,通过调节菌群结构、降低炎症反应来提高大鼠肠道健康状况。

This study was conducted to explore the effects of hesperidin and rosmarinic acid combination on ileal morphology,microbiota structure and inflammation response of rats challenged with lipopolysaccharide(LPS).Thirty-two specific-pathogen-free(SPF)Sprague-Dawley(SD)rats aged 21 days were randomly divided into four groups:LPS group,Hes group,RA group,Hes×RA group,and each group consisted of four replicates with two rats per replicate,half male and female.All rats were fed a standard basal diet.Rats in Hes group were orally gavaged 300 mg/kg BW hesperidin daily,rats in RA group were orally gavaged 20 mg/kg BW rosmarinic acid daily,rats in Hes×RA group were orally gavaged 150 mg/kg BW hesperidin+10 mg/kg BW rosmarinic acid daily,and rats in LPS group were given equal volume of sterile saline.Gavage treatment was carried out for 14 days during the trial period.On the 13th day,all rats were injected intraperitoneally with 500μg/kg BW LPS solution to construct acute intestinal injury model.The results showed that the ileal villus height and villus length/crypt depth were significantly increased in Hes×RA group compared with LPS group(P<0.05).High-throughput sequencing results showed that the richness and diversity of ileal digesta microbiota were no significant changes in Hes×RA group compared with LPS group(P<0.05).Compared with LPS group,the relative abundance of Lactobacillus in ileal digesta was significantly increased(P<0.05),while the relative abundances of Actionbacteria,Proteobacteria,Corynebacterium and Streptococcus were significantly reduced in Hes×RA group(P<0.05).Compared with LPS group,the contents of acetate,propionate and total short chain fatty acids in ileal digesta were significantly increased in Hes×RA group(P<0.05).Compared with LPS group,the myeloperoxidase(MPO)activity and the interleukin 6(IL-6)content in ileal mucosa were significantly reduced(P<0.05),while the interleukin 10(IL-10)content was significantly increased in Hes×RA group(P<0.05).Compared with LPS group,the mRNA relative expression levels of Claudin-1 and Occludin were significantly increased in Hes×RA group(P<0.05);in addition,the mRNA relative expression level of zonula occludens-1(ZO-1)trended to increase in Hes×RA group(P=0.064).Compared with LPS group,the mRNA relative expression levels of interleukin-1 receptor associator kinase 1(IRAK1)and tumor necrosis factor receptor associated protein 6(TRAF6)were significantly reduced in Hes×RA group(P<0.05).In conclusion,the combination of hesperidin and rosmarinic acid can alleviate the ileal morphology damage of rats which is caused by LPS,and improve the intestinal health by modulating microbiota structure and reducing inflammation response.