基础医学与临床 ›› 2019, Vol. 39 ›› Issue (10): 1474-1482.

• 研究论文 • 上一篇    下一篇

不同方式移植骨髓间充质干细胞治疗急性重症胰腺炎大鼠的疗效对比

傅明杰1,赵赫1,李志涛1,卢安卫1,蔡洪流1,董汉光2   

  1. 1. 浙江大学医学院附属第一医院
    2. 山东大学齐鲁医院(青岛) 普外科
  • 收稿日期:2018-11-12 修回日期:2019-01-27 出版日期:2019-10-05 发布日期:2019-09-25
  • 通讯作者: 傅明杰 E-mail:1516074@zju.edu.cn

Comparison of different methods of transplantation of bone marrow mesenchymal stem cells for acute severe pancreatitis in rats

  • Received:2018-11-12 Revised:2019-01-27 Online:2019-10-05 Published:2019-09-25
  • Contact: Ming-Jie Fu E-mail:1516074@zju.edu.cn

摘要: 目的 探索骨髓间充质干细胞(bMSCs)治疗大鼠急性重症胰腺炎(SAP)模型的高效移植方式。方法 将大鼠随机分为对照组(control)、急性重症胰腺炎组(SAP)(胰胆管内逆行注射5%硫磺胆酸钠)。SAP模型制作成功6 h后,分为尾静脉骨髓间充质干细胞移植组(tail vein)、腹腔注射移植组(intraperitoneal)和尾静脉-腹腔注射联合移植组(tail vein + intraperitoneal),每组18只。在移植后第24、48和72 h分别安乐死6只大鼠,收集胰腺组织及血清,使用ELISA试剂盒测定血清中抗炎因子IL-10及炎性因子IL-1β、TNF-α和IL-6的水平;HE染色后光学显微镜下观察胰腺组织病理和第72 h后各组大鼠小肠黏膜及肺泡组织病理改变;冰冻切片在荧光显微镜下观察DAPI标记的bMSCs在胰腺组织的分布。结果 SAP组胰腺组织有大量出血、水肿、炎性反应及坏死;与对照组相比, 3种不同方式bMSCs移植组胰腺组织的出血、水肿、炎性反应及坏死明显减少(P <0.05),IL-1β、TNF-α和IL-6水平显著降低,IL-10水平显著升高(P <0.05);尾静脉+腹腔联合移植组较单纯尾静脉或腹腔移植组胰腺组织的出血、水肿、炎性反应及坏死进一步减少(P <0.05),IL-1β、TNF-α和IL-6水平进一步降低(P <0.05),IL-10的水平进一步升高(P <0.05);DAPI标记的bMSCs在移植组的胰腺组织内均有分布,且在尾静脉+腹腔联合移植组观察到更多的荧光分布;72 h后可见移植组小肠黏膜及肺泡组织完整性较SAP组明显好转,中性粒细胞浸润及出血情况较SAP组明显减轻,相比于腹腔和单纯尾静脉移植,联合移植组情况进一步改善。结论 在大鼠SAP模型中,bMSCs 通过尾静脉+腹腔联合移植能显著抑制炎性反应,减少SAP相关性胰腺、小肠黏膜及肺损伤,其效果显著优于单纯尾静脉移植和腹腔注射移植。

关键词: 关键词:急性重症胰腺炎, 骨髓间充质干细胞, 干细胞移植。

Abstract: Objective To explore the efficient transplantation of bone marrow mesenchymal stem cells (bMSCs) in the treatment of acute severe pancreatitis (SAP) in rats. Methods Rats were randomly divided into control group and acute severe pancreatitis group (by reverse injection of 5% sodium thiocholate in the pancreatic duct). Six hours after the SAP model was successfully produced, it was divided into bone marrow mesenchymal stem cell tail vein transplantation group, intraperitoneal injection transplantation group and tail vein-peritoneal injection combined transplantation group, with 18 rats in each group. Six rats were sacrificed at 24, 48 and 72 h after transplantation, pancreatic tissue and serum were collected, and serum anti-inflammatory factor IL-10 and inflammatory factors IL-1β, TNF-α and IL-6 were measured using Elisa kit. After HE staining, the pathological changes of pancreatic tissue and the intestinal mucosa and alveolar tissue of rats in each group were observed under light microscope. The distribution of DAPI-labeled bMSCs in pancreatic tissue was observed under frozen microscope. Results There were a lot of hemorrhage, edema, inflammatory reaction and necrosis in SAP group. Compared with the control group, hemorrhage, edema, inflammatory reaction and necrosis of pancreatic tissue were significantly decreased in different ways of bMSCs transplantation group(P <0.05), IL-1β, TNF. -α and IL-6 levels were significantly decreased, IL-10 levels were significantly increased (P <0.05);Compared with tail vein transplantation group and intraperitoneal injection transplantation group, hemorrhage, edema, inflammatory response and necrosis were further reduced in the vein + intraperitoneal combined transplantation group (P <0.05), IL -1β, TNF-α and IL-6 levels were further decreased (P <0.05), IL-10 levels were further increased (P <0.05); DAPI-labeled bMSCs were distributed in the pancreatic tissue of the transplanted group, and More fluorescence distribution was observed in the vein + intraperitoneal transplantation group. After 72 hours, the integrity of intestinal mucosa and alveolar tissue in the transplanted group were significantly improved(compared with the SAP group). The neutrophil infiltration and hemorrhage were significantly reduced compared with the SAP group, Compared with intraperitoneal transplantation and simple tail vein transplantation, the combined transplantation group was further improved. Conclusions bMSCs can significantly inhibit inflammatory response and reduce SAP related pancreatic injury, lung injury and small intestinal mucosa injury through tail vein + intraperitoneal transplantation in SD rats of SAP model, and the effect is significantly better than simple tail vein transplantation or intraperitoneal injection.

Key words: Key words: acute severe pancreatitis, bone marrow mesenchymal stem cells, stem cell transplantation.