设为首页
加入收藏
求医中医药网
www.qe.cn
站内搜索
目的观察中药清胰汤(CM)及双歧杆菌合剂(BM)对犬急性坏死性胰腺炎(ANP)肠道细菌移位(BT)的影响.
方法杂种犬31只，分对照组(n=7)、ANP组(n=8)、中药治疗组(CM，n=8)和双歧杆菌合剂治疗组(BM，n=8). 对照组犬仅行剖腹术；经主胰管注入5%牛磺胆酸钠(0.5mL/kg)和胰蛋白酶(3000u/kg)复制ANP模型，CM组和BM组犬术后每日分别经胃管灌服中药清胰汤及双歧杆菌合剂.
结果与ANP组比较，CM及BM组肠粘膜损害明显减轻；肠粘膜菌群中大肠杆菌、类杆菌数量减少，双歧杆菌、乳杆菌数量显著增加(P＜0.05)，肠道微生态趋于平衡；CM组脏器细菌移位率减少50%，BM组减少37.5%，两组移位细菌数量减少10～40倍；两组血培养阳性率均由100%降为37.5%；两组血中内毒素水平下降1～2倍，淀粉酶水平下降2～3倍，磷脂酶A2活性亦显著下降，并均于d7后接近正常；肠通透性下降.
结论中药清胰汤及双歧杆菌合剂具有减轻ANP后肠粘膜损害，调节肠道菌群微生态平衡、保护肠屏障功能，从而减少BT致肠源性感染的作用.
中国图书馆分类号R576
Effect of Chinese medicine “Qing Yi Tang” and bifidobacterium mixture on intestinal bacterial translocation following acute necrotizing pancreatitis
WU Cheng-Tang1, LI Zhan-Liang2, HUANG Xiang-Cheng1 and ZHANG Zhi-Ling3
1Department of General Surgery, Nanfang Hospital, First Military Medical University, Guangzhou 510515, Guangdong Province, China
2304th Hospital of People's Liberation Army, Beijing 100037, China
3Surgical Faculty of Traditional Chinese Medicine Department, First Military Medical University, Guangzhou 510515, Guangdong Province, China
Subject headingspancreatitis/therapy; bacterial translocation; Qing Yi Tang; bifidobacterium mixture
Abstract
AIMTo observe the effect of Chinese medicine “Qing Yi Tang ” and bifidobacterium mixture on intestinal bacterial translocation (BT) in acute necrotizing pancreatitis (ANP).
METHODSThe dogs were randomly divided into four groups. Control group (n=7), received laparotomy only; ANP group (n=8), ANP model was induced by injection of 0.5mL/kg 5% sodium taurocholate with 3000U/kg trypsin into the pancreatic duct; Chinese medicine group (CM, n=8) and bifidobacterium mixture group (BM, n=8), In CM and BM groups, dogs were fed every day with “Qing Yi Tang” and bifidobacterium mixture (containing bifidobacteria 109cfu/g, lactobacilli 108cfu/g and enterococci 108cfu/g), respectively.
RESULTSIn CM and BM groups, histologic and ultrastructural damages in pancreas and ileal mucosa were much milder than those of ANP group. In ANP group, there was a significant increase of E.coli and bacteroids and a significant decrease of bifidobacteria, lactobacilli and enterococci in the intestinal mucosa and cecal content, while in CM and BM groups, these changes were alleviated significantly (P＜0.05). As compared with those of ANP group, the BT rate was reduced by 50%in CM group and 37.5%in BM group, and the counts of translocated bacteria were decreased 10-40times as well. Intestinal permeability and the levels of LPS, AMY and PLA2 were decreased 2-3 times significantly in CM and BM groups (P＜0.05).
CONCLUSIONBoth Chinese medicinal formula “Qing Yi Tang” and bifidobacterium mixture showed their protective effects on gut barrier function by alleviating the damage of intestinal mucosa and microecologic disturbance following acute pancreatitis. As a result, the chances of BT and enterogenic infection declined. These two preparations might be promising in the prophylaxis and treatment of infection complicating ANP.
0引言
急性坏死性胰腺炎(ANP)后感染发生率高达40%，是导致死亡的主要因素［1］. 研究证实，ANP后继发的胰腺感染主要是肠道细菌移位(BT)所致［2］，而细菌移位的发生又取决于肠粘膜屏障的功能状态. 目前临床上尚无特效方法用于保护肠屏障功能、阻止ANP后肠道细菌移位的发生. 本研究观察了中药清胰汤和肠道微生态调理剂(双歧杆菌合剂)对ANP犬的肠屏障功能和肠道细菌移位的影响，结果报道如下.
1材料和方法
1.1材料杂种犬31只，体重15kg±2kg. 随机分为4组：对照组(Contr, n=7)，仅行剖腹手术；ANP组(8只)，剖腹后经主胰管内注入5%牛磺胆酸钠(0.5mL/kg)和胰蛋白酶(3000u/kg)，制作ANP模型，术后根据需要输液；中药治疗组(CM，8只)，每日经胃管灌服清胰汤20mL/kg，连用7d，清胰汤组成：大黄(后下)、南柴胡、白芍各24g，黄芩、胡黄连、元胡、木香、芒硝(冲服)各18g，用水煎成300mL；双歧杆菌合剂治疗组(BM，8只)，每天经胃管灌入双歧杆菌合剂(0.5g/kg，每g用生理盐水1mL溶解)，其组成：双歧杆菌(109CFU/g)，嗜酸乳杆菌(108CFU/g)，肠球菌(108CFU/g). d7活杀.
1.2观察指标
1.2.1肠通透性分别于手术后d1,d2,d4,d7各给犬灌服5%甘露醇和10%乳果糖混合液100mL，收集6h尿，采用改良Dumas法［3］，对尿中乳果糖和甘露醇作肟化和硅烷化处理后，气相色谱法测定其含量，以乳果糖/甘露醇比率(L/M)表示肠通透性大小［4］.
1.2.2肠道微生态分析处死后无菌操作剪取空、回、盲肠标本，立即置于无菌试管用生理盐水边漂洗，边通入高纯氮(99.99%)，反复漂洗3次，然后取出组织，匀浆，接种至E.coli、肠球菌、双歧杆菌、类杆菌和乳杆菌五种选择性平板，需氧菌37℃培养24h，厌氧菌37℃厌氧箱中培养48h，作细菌计数.
1.2.3血浆内毒素(LPS)，淀粉酶(AMY)和血清磷脂酶A2(PLA2)活性术后d1,d2,d4,d7采血，LPS采用鲎试剂基质显色定量法、AMY应用碘-淀粉法，PLA2应用放射免疫法进行测定.
1.2.4肠道细菌移位活杀后无菌操作剪取肝、胰、脾、肺、肾及肠系膜淋巴结(MLN)0.1g～0.2g，匀浆后接种于需氧菌、厌氧菌平板，培养，计数.
1.2.5病理观察取胰腺及回肠组织标本，作光镜及透射电镜观察.
1.2.6动物死亡率.
统计学处理数据作F检验，结果以±s表示.
2结果
2.1病理学结果光镜观察：ANP组胰腺腺泡有大片坏死，出血，大量炎性细胞浸润；肠绒毛顶端部分破损，粘膜上皮内有细菌侵入. CM组胰腺腺泡完整，间质充血水肿，炎性细胞浸润，无坏死，出血；肠粘膜上皮基本完整，未见细菌侵入. BM组胰腺腺泡有个别小坏死灶，间质亦见充血水肿，炎性细胞浸润；肠粘膜上皮尚完整，未见细菌侵入. 电镜观察：ANP组胰腺腺泡粗面内质网扩张，线粒体退变，结构模糊；肠上皮微绒毛缩短，部分脱落. CM组和BM组胰腺腺泡内质网仅轻度扩张，线粒体无明显肿胀；肠粘膜微绒毛尚整齐，胞内细胞器结构基本正常. 对照组犬胰腺、回肠粘膜组织结构光镜及电镜下均未见异常.
2.2脏器细菌移位发生率ANP组肠系膜淋巴结(MLN)培养全部阳性，其余脏器细菌培养结果：胰、肝(7/8)，肾、肺(6/8)，脾(4/8). 中药组细菌培养结果：MLN 4/8，胰、肺、脾(3/8)，肝、肾(2/8). 微生态组细菌培养结果：MLN 5/8，胰、肺、脾(3/8)，肝1/8、肾2/8. 与ANP组比较，中药组细菌移位发生率减少50%(P＜0.05)，微生态组减少37.5%(P＜0.05)，两组脏器移位细菌数量减少了10～40倍(P＜0.05或P＜0.01，表1).
表1各脏器移位细菌计数CFUlog/g,
