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. Results ①In 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).③ The 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.

目的 探讨血必净注射液对SAP大鼠TLR4信号通路介导肠黏膜屏障功能障碍的影响。方法 24只SD大鼠随机分成空白组(n=8)、对照组(n=8)和治疗组(n=8)。对照组和治疗组用4.5%牛磺胆酸钠溶液胆胰管逆行注射制备SAP模型,空白组采用等量生理盐水逆行注射。治疗组在造模3 h后经鼠尾静脉注射血必净注射液(3 mL/kg)。三组大鼠造模后观察24 h,然后处死取胰腺和小肠组织送病理检查,采用荧光RT-PCR技术检测TLR4和NF-κB表达水平,采用ELSIA法检测血清TNF-α、IL-6、淀粉酶(AMS)及二胺氧化酶(DAO)水平,比较三组大鼠各项指标。结果 对照组和治疗组小肠组织TLR4和NF-κB表达以及血清TNF-α、IL-6、AMS及DAO水平均高于空白组(P>0.05),治疗组小肠组织TLR4和NF-κB表达以及血清TNF-α、IL-6、AMS及DAO水平低于对照组(P＜0.05)。结论 血必净注射液通过干预SAP大鼠TLR4信号通路,降低小肠组织TLR4和NF-κB的表达,减轻小肠组织的炎症反应,对肠黏膜屏障具有一定的保护作用。

Objective To investigate the effect on intestinal mucosal barrier dysfunction (IBF) of Xuebijing injection mediated by Toll-like receptor 4 (TLR4) signal pathway in rats of severe acute pancreatitis (SAP). Methods 24 Sprague Dawley (SD) rats were randomly divided into Sham group (n=8), control group (n=8) and treatment group (n=8). The SAP model was established by retrograde injection of 4.5% sodium taurocholate into the biliopancreatic duct in control group and treatment group, while control group was injected with the same amount of saline. In treatment group, Xuebijing injection (3 mL/kg) was injected via tail vein after 3h of modeling. All rats were monitored and sacrificed after 24 hours of modeling. Samples of pancreas and intestine were collected for pathologic determination. A fluorescent RT-PCR was used to determine the expression of TLR4 and NF-κB of small intestine. The serum levels of TNF-α, IL-6, amylase (AMS) and diamine oxidase (DAO) were measured by using ELISA. All parameters of three groups were compared. Results The expression of TLR4 and NF-κB of small intestine in control group and treatment group were higher than it in control group (P<0.05), as well as the serum levels of TNF-α, IL-6, AMS and DAO (P<0.05). The expression of TLR4 and NF-κB of small intestine in treatment group were lower than it in control group (P<0.05), as well as the serum levels of TNF-α, IL-6, AMS and DAO (P<0.05). Conclusion Xuebijing injection may not only reduce the expression of TLR4 and NF-κB of small intestine, but also alleviate the inflammation reaction of small intestine by interfering with TLR4 signal pathway, which may have a protective effect on intestinal mucosal barrier in SAP rats.

目的 探究血必净联合乌司他丁治疗重症急性胰腺炎的疗效及对血清细胞因子水平的影响。方法 选择2016年2月—2017年3月我院收治的重症急性胰腺炎患者88例,随机将其为两组,各44例。对照组采用乌司他丁治疗,在此基础上给予观察组血必净治疗,比较2组临床疗效、血清细胞因子及药物不良反应。结果 观察组总有效率较对照组高,TNF-α、IL-6、hs-CRP水平较对照组低,差异有统计学意义(P＜0.05);观察组不良反应率略低于对照组,但差异无统计学意义(P＞0.05)。结论 血必净联合乌司他丁可提高重症急性胰腺炎临床疗效,减轻炎性反应,且不增加不良反应率,安全性较高。