严重烫伤大鼠早期胰腺外分泌功能变化的机制研究  

Study on the mechanism of early pancreatic exocrine function changes in severely scalded rats

作　　者：罗鹏 张博涵 孙佳辰 李志生 刘馨竹 杨键秋 申传安[1] Luo Peng;Zhang Bohan;Sun Jiachen;Li Zhisheng;Liu Xinzhu;Yang Jianqiu;Shen Chuan'an(Department of Burns and Plastic Surgery,the Fourth Medical Center of PLA General Hospital,Beijing 100048,China)

机构地区：[1]解放军总医院第四医学中心烧伤整形医学部,北京100048

出　　处：《中华烧伤与创面修复杂志》2023年第5期424-433,共10页Chinese Journal of Burns And Wounds

基　　金：国家自然科学基金面上项目(82072169,82272279);军队后勤科研项目保健专项(22BJZ35);军队后勤科研重大项目(ALB18J001)。

摘　　要：目的探讨严重烫伤大鼠早期胰腺外分泌功能变化的机制。方法采用实验研究方法。取80只7~8周龄雄性SD大鼠,按随机数字表法分为单纯假伤组(8只)、假伤+胆囊收缩素八肽(CCK8)组(8只)、严重烫伤+CCK8组(32只)和特重烫伤+CCK8组(32只),分别作相应处理。2组假伤大鼠伤后即刻及2组烫伤大鼠伤后1、2、3、7 d,采用改进的胰管穿刺置管法动态收集大鼠胆胰混合液(PBJ),记录1 h内分泌PBJ的体积,采用酶联免疫吸附测定(ELISA)法检测并计算PBJ中胰脂肪酶、α-淀粉酶和胰蛋白酶的含量(样本数为8);收集股静脉血,采用标准比色法检测血清中胰脂肪酶和α-淀粉酶的浓度,反映其活力(样本数为8);提取胰腺组织,采用ELISA法检测胰腺组织中白细胞介素1β(IL-1β)和IL-6水平(样本数为8),采用免疫荧光法检测胰腺组织中缺氧诱导因子1α(HIF-1α)的表达,采用苏木精-伊红染色观察胰腺组织病理学变化,采用改良Schmidt法评估(仅2组烫伤大鼠)胰腺组织损伤的严重程度(样本数为6),采用透射电子显微镜观察胰腺组织腺泡细胞超微结构。对数据行析因设计方差分析、Tukey检验、独立样本t检验和LSD检验。结果与单纯假伤组大鼠伤后即刻胰腺组织1 h内的PBJ分泌体积(0.740±0.030)mL相比,假伤+CCK8组的(0.823±0.033)mL显著升高(t=4.92,P<0.05)。与假伤+CCK8组伤后即刻相比,严重烫伤+CCK8组[(0.681±0.024)、(0.608±0.056)、(0.525±0.025)、(0.720±0.044)mL]、特重烫伤+CCK8组[(0.540±0.025)、(0.406±0.021)、(0.475±0.036)、(0.690±0.018)mL]大鼠伤后1、2、3、7 d胰腺组织1 h内的PBJ分泌体积均显著下降(P<0.05)。与严重烫伤+CCK8组相比,特重烫伤+CCK8组大鼠伤后1、2 d胰腺组织1 h内的PBJ分泌体积均显著下降(P<0.05)。相较于单纯假伤组伤后即刻,假伤+CCK8组大鼠伤后即刻PBJ中胰脂肪酶、α-淀粉酶和胰蛋白酶含量均显著升高(t值分别为4.56、3.30、4.99,P<0.05)。与假伤+CObjective To explore the mechanism of early pancreatic exocrine function changes in severely scalded rats.Methods The experimental research methods was used.Eighty male Sprague-Dawley rats aged 7-8 weeks were divided into simple sham injury group(n=8),sham injury+cholecystokinin octapeptide(CCK8)group(n=8),severe scald+CCK8 group(n=32),and extremely severe scald+CCK8 group(n=32)by the random number table,which were treated accordingly.Immediately after injury of rats in the 2 sham injury groups and 1,2,3,and 7 days after injury of rats in the 2 scald groups,the improved methods including pancreatic duct puncture and catheterization were used to dynamically collect the pancreatic-bile juice(PBJ)of rats.The PBJ secretory volume within 1 h was recorded,and the content of pancreatic lipase,α-amylase,and trypsin in PBJ was detected by enzyme-linked immunosorbent assay(ELISA),and the number of samples was 8.The femoral venous blood was collected,and the concentrations of pancreatic lipase andα-amylase in serum were detected by standard colorimetry to reflect their activity(n=8).The pancreatic tissue was extracted,and the levels of interleukin-1β(IL-1β)and IL-6 in pancreatic tissue were detected by ELISA(n=8),the expression of hypoxia-inducible factor 1α(HIF-1α)in pancreatic tissue was detected by immunofluorescence method,and the histopathological changes in pancreatic tissue were observed by hematoxylin-eosin staining,the severity of pancreatic tissue injury in the 2 scald groups was evaluated by modified Schmidt method(n=6),and the ultrastructure of acinar cells in pancreatic tissue was observed by transmission electron microscopy.Data were statistically analyzed with analysis of variance for factorial design,Tukey test,independent sample t test,and least significant difference test.Results Compared with the PBJ secretory volume(0.740±0.030)mL in the pancreatic tissue of rats in simple sham injury group within 1 h immediately after injury,the(0.823±0.033)mL in sham injury+CCK8 group was significantly increase

关 键 词：烧伤 胰腺外分泌功能不全 胰脂肪酶 Α淀粉酶 胆囊收缩素 

分 类 号：R644[医药卫生—外科学]