安石榴苷改善小鼠细菌性肠炎及调节肠道菌群的实验研究  

Experimental study on improvement of bacterial enteritis and regulation of intestinal flora in mice by punicalagin

作　　者：黄淑芸 许萍 洪宗元[1] 钟树志[1] HUANG Shu-yun;XU Ping;HONG Zong-yuan;ZHONG Shu-zhi(Department of Histology and Embryology,Wannan Medical College,Wuhu 241002,China)

机构地区：[1]皖南医学院组织胚胎学教研室,安徽芜湖241002

出　　处：《中草药》2022年第10期3044-3052,共9页Chinese Traditional and Herbal Drugs

基　　金：安徽省高校自然科学重大项目(KJ2020ZD56);大学生科研资助金项目(WK2020S59,WK2021XS61)。

摘　　要：目的研究安石榴苷对细菌性肠炎的改善作用及对肠道菌群的影响。方法45只BALB/c小鼠采用致病性大肠杆菌O 101建立细菌性肠炎模型,给予安石榴苷或头孢克肟干预7 d,给药过程中观察各组小鼠的体征并计算每天的疾病活动指数(disease activity index,DAI)与体质量变化率;给药结束后取肠组织及肠道内容物,苏木素-伊红(HE)染色法观察小鼠肠组织的病理改变;免疫组化法检测小鼠小肠组织黏膜中闭锁连接蛋白-1(zonula occludens-1,ZO-1)和紧密连接蛋白(Occludin)的表达;测定小鼠肠组织髓过氧化物酶(myeloperoxidase,MPO)活性;qRT-PCR法检测小鼠肠壁炎症相关因子核苷酸结合寡聚化结构域蛋白2(nucleotide binding oligomerization domain 2,NOD2)、核因子-κB p65(nuclear factor-κB p65,NF-κB p65)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白细胞介素-6(interleukin-6,IL-6)mRNA的表达;ELISA法检测小鼠血清中脂多糖(lipopolysaccharide,LPS)、TNF-α和IL-6水平;16S rDNA高通量测序法检测小鼠肠道菌群多样性和丰度。结果与模型组比较,安石榴苷组小鼠DAI评分显著降低(P<0.01);小肠黏膜充血、水肿明显减轻;小肠上皮中ZO-1和Occludin蛋白表达显著增多(P<0.01);肠壁MPO活性显著降低(P<0.01);肠壁NOD2、NF-κB p65、TNF-α和IL-6 mRNA表达水平显著降低(P<0.01);血清LPS、TNF-α和IL-6水平显著减少(P<0.01);肠道厚壁菌门(Firmicutes)、毛螺菌科(Lachnospiraceae)和Clostridiales-unclassified相对丰度显著升高(P<0.01),变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、Muribaculaceae和脱硫弧菌科(Desulfovibrionaceae)菌群相对丰度显著降低(P<0.05、0.01)。结论安石榴苷能够改善致病性大肠杆菌O 101诱导的肠炎,减轻小鼠肠壁的炎症反应,保护肠黏膜屏障,其机制可能与调节肠道菌群有关。Objective To study the improvement effect of punicalagin on bacterial enteritis and its influence on intestinal flora.Methods Forty-five BALB/c mice were ig pathogenic Escherichia coli O 101 to establish a bacterial enteritis model,and were given punicalagin or cefixime for 7 d of intervention.During the administration,signs of mice in each group were observed,daily disease activity index(DAI)and body weight change rate were calculated;Intestinal tissue and intestinal contents were collected after administration,and hematoxylin-eosin(HE)staining was used to observe the pathological changes of intestinal tissue in mice;Expressions of tight junction protein zonula occludens-1(ZO-1)and Occludin in small intestine were detected by immunohistochemistry;Activity of myeloperoxidase(MPO)in intestinal tissue of mice was measured;Inflammation related factors nucleotide binding oligomerization domain 2(NOD2),nuclear factor-κB p65(NF-κB p65),tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)mRNA expressions in intestinal were detected by qRT-PCR;Lipopolysaccharide(LPS),TNF-α,IL-6 levels in serum were determined by ELISA;Diversity and abundance of intestinal flora in mice were detected by 16S rDNA high throughput sequencing.Results Compared with model group,DAI score of mice in punicalagin group was significantly decreased(P<0.01);Congestion and edema of small intestinal mucosa were significantly alleviated;Expressions of small intestinal epithelial tight junction associated proteins ZO-1 and Occludin were significantly increased(P<0.01);MPO activity in intestinal wall was significantly reduced(P<0.01);NOD2,NF-κB p65,TNF-αand IL-6 mRNA expressions in intestinal wall were decreased significantly(P<0.01);LPS,TNF-αand IL-6 levels in serum were significantly decreased(P<0.01);Relative abundances of Firmicutes,Lachnospiraceae and Clostridiales-unclassified were significantly increased(P<0.01),relative abundances of Proteobacteria,Bacteroidetes,Muribaculaceae and Desulfovibrionaceae were significantly decreased(P<0.05,0.

关 键 词：安石榴苷 细菌性肠炎 肠道菌群 肠黏膜屏障 炎症反应 

分 类 号：R285.5[医药卫生—中药学]