中华烧伤杂志
中華燒傷雜誌
중화소상잡지
16
2011年
1期
49-53
,共5页
冯永强%王德昌%王坤%冷向锋%肖虎%郭丹凤
馮永彊%王德昌%王坤%冷嚮鋒%肖虎%郭丹鳳
풍영강%왕덕창%왕곤%랭향봉%초호%곽단봉
烧伤%肠%淋巴%细菌移位%内毒素类
燒傷%腸%淋巴%細菌移位%內毒素類
소상%장%림파%세균이위%내독소류
Burns%Intestines%Lymph%Bacterial translocation%Endotoxins
目的 了解淋巴途径在烫伤大鼠肠道细菌移位中的作用.方法 制备羰花青荧光染料CM-DIL标记的大肠埃希菌菌液(数量级109CFU/L).将60只成年雄性Wistar大鼠按照信封法随机分为烫伤组、假伤组,每组30只.2组大鼠经胃灌注已制备的标记菌液0.5 mL后,烫伤组致30%TBSA深Ⅱ度烫伤,伤后立即行液体复苏;假伤组大鼠皮肤经25℃水浴10 s模拟烫伤并同法复苏.采集2组大鼠伤后2、24、72 h(每时相点10只)的肠系膜淋巴结(MLN)、肝脏、肠系膜淋巴液(MLF)、肝静脉血标本.分别采用荧光示踪法和细菌培养法检测细菌移位情况;用鲎试剂显色基质法定量测定上述4种标本中内毒素含量,并计算MLF和肝静脉血的内毒素携带量.对实验数据行t检验或单因素方差分析.结果 (1)荧光示踪法检测显示活菌呈短棒状,液体标本中可见单体或两三个联体的可移动活菌;死菌呈不规则碎片状.假伤组伤后2 h有少量标记菌,24 h数量达高峰;烫伤组伤后2 h标记菌较多,24、72 h仍处于较高水平.烫伤组各种标本比较,MLN内标记菌最多,伤后24 h达高峰[(5872±1976)×103 CFU/g],明显多于假伤组[(216±110)×103 CFU/g,t=30.129,P=0.000],72 h有所减少但仍多于假伤组(t=4.323,P=0.000);血液内标记菌最少.(2)细菌培养法检测:假伤组120份标本中,29份细菌培养呈阳性占24.2%;烫伤组120份标本中,72份培养阳性占60.0%.2组大鼠除血液标本在各时相点均未检出活菌外,其余3种标本伤后2 h或24 h可培养出活菌.烫伤组MLN和肝脏较其余2种组织检出更多活菌;伤后24 h,该组MLN、肝脏以及MLF标本中细菌数量均明显多于假伤组(t值分别为4.353、4.354、4.965,P值均等于0.000).(3)内毒素含量:烫伤组大鼠伤后各时相点4种标本的内毒素含量均高于假伤组,其中肝脏、MLF内毒素含量伤后2 h即达高峰.此时相点烫伤组4种组织间内毒素含量比较,差异有统计学意义(F=258.47,P=0.000),其中肝脏、MLN、MLF内毒素含量较高,且明显高于假伤组(t值分别为43.378、43.123、22.423,P值均等于0.000);MLF内毒素含量约为血液中含量的9倍.(4)烫伤组伤后各时相点MLF、血液内毒素携带量均高于假伤组.结论 用CM-DIL标记细菌能够较全面反映细菌移位情况,淋巴途径在细菌移位中发挥重要作用.
目的 瞭解淋巴途徑在燙傷大鼠腸道細菌移位中的作用.方法 製備羰花青熒光染料CM-DIL標記的大腸埃希菌菌液(數量級109CFU/L).將60隻成年雄性Wistar大鼠按照信封法隨機分為燙傷組、假傷組,每組30隻.2組大鼠經胃灌註已製備的標記菌液0.5 mL後,燙傷組緻30%TBSA深Ⅱ度燙傷,傷後立即行液體複囌;假傷組大鼠皮膚經25℃水浴10 s模擬燙傷併同法複囌.採集2組大鼠傷後2、24、72 h(每時相點10隻)的腸繫膜淋巴結(MLN)、肝髒、腸繫膜淋巴液(MLF)、肝靜脈血標本.分彆採用熒光示蹤法和細菌培養法檢測細菌移位情況;用鱟試劑顯色基質法定量測定上述4種標本中內毒素含量,併計算MLF和肝靜脈血的內毒素攜帶量.對實驗數據行t檢驗或單因素方差分析.結果 (1)熒光示蹤法檢測顯示活菌呈短棒狀,液體標本中可見單體或兩三箇聯體的可移動活菌;死菌呈不規則碎片狀.假傷組傷後2 h有少量標記菌,24 h數量達高峰;燙傷組傷後2 h標記菌較多,24、72 h仍處于較高水平.燙傷組各種標本比較,MLN內標記菌最多,傷後24 h達高峰[(5872±1976)×103 CFU/g],明顯多于假傷組[(216±110)×103 CFU/g,t=30.129,P=0.000],72 h有所減少但仍多于假傷組(t=4.323,P=0.000);血液內標記菌最少.(2)細菌培養法檢測:假傷組120份標本中,29份細菌培養呈暘性佔24.2%;燙傷組120份標本中,72份培養暘性佔60.0%.2組大鼠除血液標本在各時相點均未檢齣活菌外,其餘3種標本傷後2 h或24 h可培養齣活菌.燙傷組MLN和肝髒較其餘2種組織檢齣更多活菌;傷後24 h,該組MLN、肝髒以及MLF標本中細菌數量均明顯多于假傷組(t值分彆為4.353、4.354、4.965,P值均等于0.000).(3)內毒素含量:燙傷組大鼠傷後各時相點4種標本的內毒素含量均高于假傷組,其中肝髒、MLF內毒素含量傷後2 h即達高峰.此時相點燙傷組4種組織間內毒素含量比較,差異有統計學意義(F=258.47,P=0.000),其中肝髒、MLN、MLF內毒素含量較高,且明顯高于假傷組(t值分彆為43.378、43.123、22.423,P值均等于0.000);MLF內毒素含量約為血液中含量的9倍.(4)燙傷組傷後各時相點MLF、血液內毒素攜帶量均高于假傷組.結論 用CM-DIL標記細菌能夠較全麵反映細菌移位情況,淋巴途徑在細菌移位中髮揮重要作用.
목적 료해림파도경재탕상대서장도세균이위중적작용.방법 제비탄화청형광염료CM-DIL표기적대장애희균균액(수량급109CFU/L).장60지성년웅성Wistar대서안조신봉법수궤분위탕상조、가상조,매조30지.2조대서경위관주이제비적표기균액0.5 mL후,탕상조치30%TBSA심Ⅱ도탕상,상후립즉행액체복소;가상조대서피부경25℃수욕10 s모의탕상병동법복소.채집2조대서상후2、24、72 h(매시상점10지)적장계막림파결(MLN)、간장、장계막림파액(MLF)、간정맥혈표본.분별채용형광시종법화세균배양법검측세균이위정황;용후시제현색기질법정량측정상술4충표본중내독소함량,병계산MLF화간정맥혈적내독소휴대량.대실험수거행t검험혹단인소방차분석.결과 (1)형광시종법검측현시활균정단봉상,액체표본중가견단체혹량삼개련체적가이동활균;사균정불규칙쇄편상.가상조상후2 h유소량표기균,24 h수량체고봉;탕상조상후2 h표기균교다,24、72 h잉처우교고수평.탕상조각충표본비교,MLN내표기균최다,상후24 h체고봉[(5872±1976)×103 CFU/g],명현다우가상조[(216±110)×103 CFU/g,t=30.129,P=0.000],72 h유소감소단잉다우가상조(t=4.323,P=0.000);혈액내표기균최소.(2)세균배양법검측:가상조120빈표본중,29빈세균배양정양성점24.2%;탕상조120빈표본중,72빈배양양성점60.0%.2조대서제혈액표본재각시상점균미검출활균외,기여3충표본상후2 h혹24 h가배양출활균.탕상조MLN화간장교기여2충조직검출경다활균;상후24 h,해조MLN、간장이급MLF표본중세균수량균명현다우가상조(t치분별위4.353、4.354、4.965,P치균등우0.000).(3)내독소함량:탕상조대서상후각시상점4충표본적내독소함량균고우가상조,기중간장、MLF내독소함량상후2 h즉체고봉.차시상점탕상조4충조직간내독소함량비교,차이유통계학의의(F=258.47,P=0.000),기중간장、MLN、MLF내독소함량교고,차명현고우가상조(t치분별위43.378、43.123、22.423,P치균등우0.000);MLF내독소함량약위혈액중함량적9배.(4)탕상조상후각시상점MLF、혈액내독소휴대량균고우가상조.결론 용CM-DIL표기세균능구교전면반영세균이위정황,림파도경재세균이위중발휘중요작용.
Objective To investigate the role of lymphatics in bacterial translocation from intestine of rats with burn. Methods Escherichia coli (E. coli) labeled with chloromethylbenzamidodialkylcarbocyanine (CM-DIL) were prepared. Sixty adult male Wistar rats were randomly divided into scald group and sham injury group according to the envelope method, with 30 rats in each group. Rats in both groups were gavaged with 0. 5 mL fluid containing CM-DIL-labeled E. coli. Rats in scald group were inflicted with 30% TBSA deep partial-thickness scald (verified by pathological section) and resuscitated with fluid. Rats in sham injury group were sham injured by bathing in 25 ℃ water for 10 s(verified by pathological section)and also received with fluid infusion. Mesenteric lymph node (MLN), liver, mesenteric lymph fluid (MLF), and liver vein blood (LVB) were harvested at post injury hour (PIH) 2, 24, and 72. Bacteria translocation was detected with fluorescent tracing technique and bacteria culture. The endotoxin content in above-mentioned four kinds of specimens was quantitatively determined with chromogenic substrate limulus amebocyte lysate. The carrying capacity of endotoxin in MLF and LVB was calculated. Data were processed with t test or one-way analysis of variance. Results (1) Living bacteria were in short-stick form, and they were seen moving in single or in doubles or triples in sample fluid. Dead bacteria were in irregular aggregates. Labeled bacteria in small amount were detected in sham injury group, their number peaked at PIH 24. A large amount of labeled bacteria were detected in scald group at PIH 2, which peaked at PIH 24 and decreased at PIH 72. The largest amount of labeled bacteria were found in MLN in scald group as compared to those in the other samples, and the number peaked at PIH 24 [(5872 ± 1976) × 103 CFU/g], which was obviously higher than that [(216 ± 110) × 103 CFU/g, t =30. 129, P =0.000] in sham injury group. The number of bacteria decreased at PIH 72, but it was still significantly different from that in sham injury group ( t =4. 323, P =0.000). The number of bacteria in LVB was the smallest. (2) 29 (24.2%) samples out of the 120 samples in sham injury group were positive for bacteria. 72 (60.0%) samples out of the 120 samples in scald group were positive for bacteria. No alive bacterium was detected at any time point in LVB sample in both group; the other three samples were detected with alive bacteria since PIH 2. There were more alive bacteria detected in MLN and liver as compared with the other two kinds of samples in scald group. The amount of bacteria in MLN, liver, and MLF in scald group were higher than those in sham injury group(with t value respectively 4. 353, 4. 354, 4. 965, P values all equal to 0. 000). (3) The endotoxin level in each kind of sample at each time point was obviously higher in scald group than that in sham injury group, and it peaked at PIH 2 in liver and MLF. The difference of endotoxin level among 4 kinds of samples in scald group at PIH 2 was statistically significant(F = 258.47, P = 0. 000) , and the endotoxin level was higher in liver, MLN, and MLF. They were obviously higher than those in sham injury group(with t value respectively 43. 378, 43. 123, 22. 423, P values all equal to 0. 000). The endotoxin level in MLF was 9 times of that in LVB. (4) The carrying capacity of endotoxin in LVB and MLF at each time point in scald group was higher than that in sham injury group. Conclusions CM-DIL marked bacteria can reflect the microbial translocation condition. The lymphatic route is an important pathway for bacteria translocation.
