血必净注射液对ANP大鼠肠道菌群及肠黏膜屏障功能的影响

Effect of Xuebijing injection on intestinal flora and intestinal mucosal barrier function in ANP rats

目的探讨血必净注射液对ANP大鼠肠道菌群及肠黏膜屏障功能的影响。方法40只SD大鼠随机分为空白组、假手术组、ANP组和血必净治疗组(每组10只),空白组不作任何处理,假手术组翻动十二指肠后关腹,ANP组和治疗组用4.5%牛磺胆酸钠溶液胆胰管逆行注射建模,治疗组在建模后经鼠尾静脉注射血必净注射液(3 mL/kg)。24 h后处死大鼠并采样,ELISA法测血AMS、CRP、LPS、TNF-α、IL-6、IL-1β、DAO和D-乳酸等指标,粪菌样本行16SrRNA高通量测序分析,实时定量PCR法检测5种细菌数量,病理检测胰腺和回肠组织,比较各组大鼠的指标。结果①ANP组大鼠血AMS升高,CRP、LPS、TNF-α、IL-6、IL-1β、DAO、D-乳酸水平以及胰腺、小肠病理评分均高于空白组和假手术组(P<0.001);②治疗组AMS低于ANP组,血必净可降低上述各种血清指标水平和胰腺、小肠病理评分(P<0.001);③肠道菌群微生态分析显示,血必净可改善ANP大鼠粪菌的丰富度和多样性,缩小与空白组、假手术菌种种类的差异,增加厚壁菌门菌量;治疗组乳酸杆菌、双歧杆菌和普拉梭菌的菌量高于ANP组,肠球菌和大肠埃希的菌量低于ANP组(P<0.001)。结论血必净可增加ANP大鼠肠道菌群的丰富度和多样性,增加有益菌的含量,减少内毒素和促炎因子释放,改善肠黏膜屏障功能。

Objective To investigate the effect of Xuebijing injection on intestinal flora and intestinal mucosal barrier function in ANP rats.Methods 40 SD rats were randomly divided into blank group,sham operation group,ANP group and Xuebijing treatment group(10 in each group).The sham operation group closed the abdomen after turning the duodenum.The ANP model was established by retrograde injection of 4.5%sodium taurocholate solution into the biliopancreatic duct.Xuebijing injection(3mL/kg)was injected into the tail vein of the rats in the treatment group.24 hours later,the rats were sacrificed and sampled.AMS,CRP,LPS,TNF-,il-6,il-1,DAO and d-lactic acid were measured by ELISA.The fecal bacteria samples were analyzed by 16SrRNA sequencing technique.Real-time quantitative PCR was used to detect the populations of 5 bacteria in fecal sample.The pathology of pancreas and ileum were examined,and the indexes of rats in each group were compared.Results①In ANP group,AMS was increased,levels of CRP,LPS,TNF-,il-6,il-1,DAO,d-lactic acid,pancreatic and intestinal pathology scores were higher than those in the blank group and the sham group(P<0.001).②In treatment group,AMS was lower than ANP group,and Xuebijing could reduce the levels of the above factors and scores of pancreatic and intestinal pathology(P<0.001).③The microecological results of intestinal flora showed that Xuebijing treatment could improve the richness and diversity of fecal bacteria,reduce the difference between Xuebijing group and blank group and sham operation group,and increase the quantity of firmicutes.The amount of Lactobacillus,Bifidobacteria and Clostridium prasei in the Xuebijing group was higher than that in ANP group,while the amount of enterococci and Escherichia coli was lower than that in the ANP group(P<0.001).Conclusion Xuebijing can increase the richness and diversity of intestinal flora,increase the content of beneficial bacteria,reduce the release of endotoxin and pro-inflammatory factors,and improve the intestinal mucosal barrier function in ANP rats.