肠三叶因子和黏蛋白对烧伤血清所致肠上皮细胞免疫功能变化的影响  

Effects of intestinal trefoil factor combined with mucin on immune function of burn serum treated intestinal epithelial cells

作　　者：吴修文[1] 王焕[1] 万千雪[1] 金星[1] 孙勇[1] 吴丹[1] 曹俊杰[1] 彭曦[1] 

机构地区：[1]第三军医大学西南医院全军烧伤研究所 创伤、烧伤与复合伤国家重点实验室,重庆400038

出　　处：《中华烧伤杂志》2011年第5期341-346,共6页Chinese Journal of Burns

基　　金：基金项目：国家自然科学基金（30971075）;“重大新药创制”科技重大专项（2009ZX09103-647）;创伤、烧伤与复合伤国家重点实验室自主研究课题（SKLZZ200807）

摘　　要：目的观察肠三叶因子（ITF）和黏蛋白对烧伤血清所致肠上皮细胞免疫功能变化的影响。方法（1）体外培养大鼠小肠上皮细胞株IEC-6，根据培养液添加物质不同，按照随机数字表法将细胞分为5组：①正常对照组，培养液中含10％（指体积分数，下同）小牛血清；②烧伤对照组，培养液含10％烧伤血清；③ITF＋烧伤血清组，培养液含10％烧伤血清及终浓度25ug／mLITF；④黏蛋白＋烧伤血清组，培养液含10％烧伤血清及终浓度250ug／mL黏蛋白；⑤ITF＋黏蛋白＋烧伤血清组，培养液含10％烧伤血清及终浓度25ug／mLITF、250ug／mL黏蛋白。向各组细胞加入上述培养液的同时，加入大肠杆菌菌液（1×10^8CFU／mL，200uL）。继续培养15min、30min、1h、2h、3h后，行瑞氏-吉姆萨染色，于显微镜下观察并统计黏附在细胞上的细菌数；采用锥虫蓝染色法观察并统计细胞存活率。每组每时相点样本数均为20。（2）将IEC-6细胞按照随机数字表法分为4组：烧伤对照组、ITF＋烧伤血清组、黏蛋白＋烧伤血清组、ITF＋黏蛋白＋烧伤血清组，分别同前加入相应的培养液（不加菌液）培养3、6、12、24、48h。采用放射免疫分析法测定各时相点培养上清液中TNF-a、IL-6和IL-8含量，每组每时相点样本数均为6。对实验数据行t检验。结果（1）烧伤对照组细胞各时相点细菌黏附数量较正常对照组明显增多（t值为2．947～8．149，P值均小于0．01）。与烧伤对照组比较，其余3个烧伤血清组在加菌后多数时相点细菌黏附的数量明显偏少（t值为-4．733～-2．180，P〈0．05或P〈0．01）。烧伤对照组细胞各时相点存活率与正常对照组比较均明显降低（t值为-4．126～-2．363，P值均小于0．05）。ITF＋烧伤血清组、黏蛋白＋烧伤血清组细胞存活率在部分时相点明显高于烧伤对照组（t值为2．120～3．423，P〈0．05或PObjective To observe the effect of intestinal trefoil factor （ITF） combined with mucin on immune function of intestinal epithelial cells （IEC） after being treated with burn rat serum. Methods The rat IEC-6 cell lines were divided into control group （ C, cultured in DEME medium containing 10% calf serum）, burn control group （BC, cultured in DEME medium containing 10% burn rat serum）, burn serum ＋ ITF group （B ＋ I, cultured in DEME medium containing 10% burn rat serum and 25 ug/mL ITF） , burn serum ＋ mucin group （ B ＋ M, cultured in DEME medium containing 10% burn rat serum and 250 ug/mL mucin） , and burn serum ＋ ITF ＋ mucin group （ B ＋ I ＋ M, cultured in DEME medium containing 10% burn rat serum, 25 ug/mL ITF, and 250 ug/mL mucin） according to the random number table. Meanwhile, 200 uL suspension of E. coli with density of 1 × 10^8 CFU/mL was added to each culture. At post culture minute （PCM） 15, 30 and post culture hour （PCH） 1, 2, 3, the number of bacteria adherent to IEC-6 was counted after Wright-Giemsa staining, and cell survival rate was calculated after trypan blue staining, with 20 samples in each group at each time point. （2） Other samples of IEC-6 cells without addition of E. coli were divided into BC, B ＋ I, B ＋ M, and B ＋ I ＋ M groups with the same treatment as above. The supernatant contents of IL-6, IL-8, and TNF-a were determined by radioimmunoassay at PCH 3,6, 12, 24, 48, with 6 samples in each group at each time point. Data were processed with t test. Results （ 1 ） Compared with that in C group, count of adherent bacteria to IEC-6 in BC group at each time point was significantly increased （ with t values from 2. 947 to 8. 149, P values all below 0.01 ）. Compared with those in BC group, the counts in B ＋ I, B ＋ M, B ＋ I ＋ M groups at the major time points were significantly decreased （ with t values from -4.733 to -2. 180,P 〈0.05 orP 〈0.01）. （2） Compared with that inC group, cell survival rate in BC group

关 键 词：烧伤 黏蛋白类 细菌黏附 肠三叶因子 肠上皮细胞 肠道免疫 

分 类 号：R644[医药卫生—外科学]