IL-10/JAK1/STAT3通路对SIS转归影响的体内实验研究

In vivo study on interleukin-10/JAK1/STAT3 pathway on degradation and regeneration of small intestinal submucosa

目的:探讨白细胞介素(interleukin,IL)10能否通过激活JAK1/STAT3通路抑制小肠黏膜下基质(small intestinal submucosa,SIS)收缩,为SIS更好地修复组织功能缺损寻找新思路。方法:大鼠背部植入2.0 cm×2.0 cm的4层SIS补片。60只SD大鼠随机分为4组(n=15):IL-10组、S3I-201(IL-10下游通路阻断剂)组、IL-10+S3I-201组和生理盐水对照组。术后1、4、8周每组各处死5只。通过毫米尺测量SIS面积后计算其收缩率。HE染色及免疫组织化学法检测SIS中信号转导及转录活化因子(signal transducers and activator of transcription,STAT)3、基质金属蛋白酶(matrix metalloproteinase,MMP)2、血管内皮生长因子(vascular endothelial growth factor,VEGF)和α平滑肌肌动蛋白(alphasmooth muscle actin,α-SMA)表达与含量。结果 :术后第1周各组SIS收缩率、STAT3、MMP2、VEGF及α-SMA表达无统计学差异(P>0.05)。与对照组比较,第4、8周IL-10可明显抑制SIS收缩、增加STAT3通路激活数、促进SIS降解及新生血管生成、抑制肌成纤维细胞生成(P<0.05)。S3I-201可对抗IL-10的以上作用(P<0.05)。经测量,术后8周IL-10组、S3I-201组和IL-10+S3I-201组收缩率分别为(39.8±1.0)%、(84.3±0.8)%、(69.8±0.6)%。量化计分,8周时3组STAT3含量分别为22.3±1.4、9.2±0.9、15.4±1.0;MMP2含量分别为25.0±1.0、9.4±0.8、15.0±0.6;VEGF含量分别为17.5±0.4、4.7±0.4、9.1±0.5;α-SMA含量分别为31.4±0.5、85.6±0.7、47.8±0.8。结论 :IL-10通过激活JAK1/STAT3通路,促进外源性胶原基质降解、新生血管生成及减少肌成纤维细胞生成,发挥抑制SIS修复组织收缩的作用。

Objective To investigate whether interleukin-10（IL-10） restrains the shrinkage of small intestinal submucosa（SIS） repair tissue by activating the JAK1/STAT3 pathway. Methods A four-ply SIS with 2.0 cm ×2.0 cm was implanted into the back of SD rat. Sixty rats were randomly divided into 4 groups（n=15）： IL-10 group, group of S3I-201 which is the inhibitor of downstream pathway of IL-10, IL-10＋S3I-201 group and the saline control group. Five rats each group were sacrificed at the 1st, 4th and 8th week after implantation. The shrinkage rate of SIS repair tissue was measured by millimeter. HE staining and immunohistochemistry were done to detect the expression of STAT3, MMP2, VEGF and α-SMA of SIS by quantitative score. Results There was no obvious difference of shrinkage rates and the scores of STAT3, MMP2, VEGF and α-SMA at the 1st week after implantation（P〉0.05）. IL-10 inhibited the shrinkage of SIS repair tissue and the formation of myofibroblast more significantly at the 4th and 8th weeks（P〈0.05） when compared with saline group. The increase in both the number of activated STAT3 pathway and the degradation of SIS with angiogenesis was found in IL-10 group also at the 4th and 8th weeks（P〈0.05）. All these change were not present while using S3I-201（P〈0.05）. The shrinkage rates of SIS repair tissue were（39.8 ±1.0）%,（84.3±0.8）%,（69.8± 0.6）% at 8th week after implantation in the IL-10 group, S3I-201 group and IL-10 ＋S3I-201 group respectively. The scores of STAT3 in these three groups were（22.3±1.4, 9.2±0.9, 15.4±1.0） and（25.0±1.0, 9.4±0.8, 15.0± 0.6） for MMP2,（17.5±0.4, 4.7±0.4,9.1±0.5） for VEGF and（31.4±0.5, 85.6±0.7, 47.8±0.8） for α-SMA. Conclusions IL-10 can play a role in restrain of the shrinkage of SIS repair tissue by activating the JAK1/STAT3 pathway which induces the degradation of exogenous colla-gen matrix and production of angiogenesis with decrease in myofibroblasts. Therefore, using IL-10 might reduce the shrinkage of SIS repair tissue.