目的: 研究严重烫伤MODS小鼠胰岛β细胞功能下降与胰高血糖素样肽1/血糖（GLP-1/GLU）比值关系并探讨其机理,从改善肠道分泌GLP-1功能新角度,为防治严重创伤MODS胰岛β细胞功能下降提供实验依据。方法: 90只SPF级的昆明小鼠随机分为对照组（10只）、单纯烫伤组（30只）和MODS组（50只）,MODS组制作为MODS小鼠（30％体表面积III度烫伤+腹腔注射内毒素）,单纯烫伤组仅造成30％体表面积III度烫伤,对照组假烫伤。取对照组小鼠采血,单纯烫伤组及MODS组伤后1天和3天均分别随机抓取10只小鼠进行采血,伤后5天将仍然存活的小鼠全部采血。测定小鼠GLU、血浆胰岛素、GLP-1和二胺氧化酶（DAO）水平。采用稳态模式评估β细胞功能（HOMA-β）,计算并观察胰岛β细胞功能指数HOMA-β和血浆GLP-1/GLU比值变化,计算HOMA-β指数与血浆GLP-1/GLU比值相关系数。取小鼠回肠组织进行GLP-1免疫组化检测,得出肠道GLP-1的表达水平,计算并观察肠道GLP-1分泌相对血糖（肠道GLP-1/GLU）水平变化。计算小鼠血浆GLP-1/GLU与肠道GLP-1/GLU相关系数、肠道GLP-1/GLU与血浆DAO相关系数。对数据行单因素方差分析及Bonferroni检验、Kruskal-Wallis及Bonferroni检验、Spearman秩相关性分析。结果: 严重烫伤MODS小鼠伤后1天、3天、5天 的HOMA-β指数、血浆GLP-1/GLU比值以及肠道GLP-1/GLU比值均明显低于对照组及单纯烫伤组同一时间点的水平;小鼠HOMA-β指数与血浆GLP-1/GLU比值呈正相关（r=0.685,P<0.001）;小鼠血浆GLP-1/GLU与肠道GLP-1/GLU呈正相关（r=0.851,P<0.001）;小鼠肠道GLP-1/GLU比值与DAO呈负相关（r=-0.732,P<0.001）。结论: 严重烫伤MODS小鼠存在胰岛β细胞功能下降及血浆GLP-1/GLU比值降低,血浆GLP-1水平相对不足与胰岛β细胞功能下降有关,肠道损害导致GLP-1分泌相对不足。保护严重烫伤MODS小鼠肠道细胞,提高血液GLP-1/GLU比值,可作为改善胰岛β细胞功能下降的重要措施。
Abstract
Objective To study the correlation between the reduction of islet β-cell function and the ratio of glucagon-like peptide 1/glucose (GLP-1/GLU) and explore its mechanism in severely scalded MODS mice so as to provide experimental basis for the prevention and treatment of the reduction of islet β-cell function in severely scalded MODS from a new perspective of improving the function of GLP-1 secretion in intestine. Methods 90 Kunming mice of Specific Pathogen Free (SPF) grade were randomly divided into three groups: 10 mice as the control group, 30 mice as simple scald group and 50 mice as MODS group. The mice in MODS group were made into MODS mice with 30% total body surface area Ⅲ degree scald and intraperitoneal injection of LPS. The mice in simple scald group were only inflicted with 30% total body surface area Ⅲ degree scald, and mice in the control group were sham scalded. Blood samples of mice in the control group were collected. At 1 day and 3 days after injury, 10 mice were randomly captured respectively from simple scald group and MODS group, and blood samples were collected. At 5 days after injury, blood samples of all surviving mice in scald group and MODS group were collected. The levels of GLU, plasma insulin, GLP-1 and diamine oxidase (DAO) of mice were determined. Homeostasis model assessment of β-cell function (HOMA-β) was applied, and then the index of islet β-cell function (HOMA-β) and the ratio of plasma GLP-1/GLU were calculated and observed. The correlation coefficient between HOMA-β and plasma GLP-1/GLU was calculated. The ileum tissues of mice were collected to observe the expression of GLP-1 by immunohistochemistry, and then the levels of intestinal GLP-1 secretion relative to blood glucose (intestinal GLP-1/GLU) were calculated and observed. The correlation coefficient between plasma GLP-1/GLU and intestinal GLP-1/GLU, and the correlation coefficient between intestinal GLP-1/GLU and plasma DAO were calculated. Data were statistically analyzed with one-way analysis of variance and Bonferroni test, Kruskal-Wallis and Bonferroni test, and Spearman rank correlation analysis. Results HOMA-β, plasma GLP-1/GLU and intestinal GLP-1/GLU of severely scalded MODS mice at 1, 3 and 5 days after injury were significantly lower than those of mice in the control group and simple scald group at the same time point (P<0.05). HOMA-β was positively correlated with plasma GLP-1/GLU of mice (r=0.685, P<0.001). Plasma GLP-1/GLU was positively correlated with intestinal GLP-1/GLU (r=0.851, P<0.001). Intestinal GLP-1/GLU was negatively correlated with DAO (r=-0.732, P<0.001). Conclusion The islet β-cell function and the ratio of GLP-1/GLU were decreased in severely scalded MODS mice. Insufficiency of plasma GLP-1 relative to blood glucose was correlated with the reduction of islet β-cell function. Intestinal damage led to relatively insufficient GLP-1 secretion. Protecting intestinal cells to increase the ratio of plasma GLP-1/GLU in severely scalded MODS mice can be used as an important measure to improve the reduction of islet β-cell function.
关键词
严重烫伤 /
MODS /
胰岛β细胞功能下降 /
GLP-1/GLU /
炎症因子
Key words
severe scalds /
MODS /
reduction of islet β-cell function /
GLP-1/GLU /
inflammatory cytokines
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基金
长沙市中医医院（长沙市第八医院）院级科研项目（2022008）
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