IL-10、TGF-β1在小鼠深静脉血栓中的时序性变化  

Time-Dependent Sequential Changes of IL-10 and TGF-β1 in Mice with Deep Vein Thrombosis

作　　者：吴娟娟 黄俊杰[1,2,3] 张煜 卓嘉英 陈刚 杨舒涵 赵蕴琦 范琰琰[1,2,3] WU Juan-juan;HUANG Jun-jie;ZHANG Yu;ZHUO Jia-ying;CHEN Gang;YANG Shuhan;ZHAO Yun-qi;FAN Yan-yan(Department of Forensic Medicine,Wenzhou Medical University,Wenzhou 325035,Zhejiang Province,China;Forensic Center,Wenzhou Medical University,Wenzhou 325035,Zhejiang Province,China;Institute of Forensic Science,Wenzhou Medical University,Wenzhou 325035,Zhejiang Province,China)

机构地区：[1]温州医科大学法医学系,浙江温州325035 [2]温州医科大学司法鉴定中心,浙江温州325035 [3]温州医科大学司法鉴定科学技术研究所,浙江温州325035

出　　处：《法医学杂志》2024年第2期179-185,共7页Journal of Forensic Medicine

基　　金：浙江省自然科学基金资助项目(LY20H150006)。

摘　　要：目的检测小鼠深静脉血栓发展过程中白细胞介素-10(interleukin-10,IL-10)和转化生长因子-β1(transforming growth factor-β1,TGF-β1)的表达变化,探讨其在血栓形成时间推断中的应用价值。方法对实验组小鼠建立下腔静脉结扎模型,分别于术后1 d、3 d、5 d、7 d、10 d、14 d、21 d过量麻醉处死小鼠,在结扎点的下方提取形成血栓的下腔静脉段。对照组小鼠不进行结扎,提取与实验组相同部位的正常下腔静脉段作为对照样本。应用免疫组织化学染色、蛋白质印迹法和real-time qPCR检测IL-10和TGF-β1在血栓形成期间的表达变化。结果免疫组织化学染色结果显示,IL-10主要表达于血栓中的单核细胞,TGF-β1主要表达于血栓中的单核细胞和成纤维样细胞。蛋白质印迹法以及real-time qPCR检测结果显示,各实验组IL-10和TGF-β1的表达水平均高于对照组。IL-10的mRNA及蛋白水平分别于术后7 d和10 d达到峰值,术后7 d的mRNA表达量是对照组的(4.72±0.15)倍,术后10 d的蛋白表达量是对照组的(7.15±0.28)倍。TGF-β1的mRNA及蛋白水平分别于术后10 d和14 d达到峰值,术后10 d的mRNA表达量是对照组的(2.58±0.14)倍,术后14 d的蛋白表达量是对照组的(4.34±0.19)倍。结论深静脉血栓演变期间IL-10和TGF-β1呈现时序性表达变化,IL-10和TGF-β1的表达水平可以为血栓形成时间推断提供参考依据。Objective To detect the expression changes of interleukin-10(IL-10)and transforming growth factor-β1(TGF-β1)during the development of deep vein thrombosis in mice,and to explore the application value of them in thrombus age estimation.Methods The mice in the experimental group were subjected to ligation of inferior vena cava.The mice were sacrificed by excessive anesthesia at 1 d,3 d,5 d,7 d,10 d,14 d and 21 d after ligation,respectively.The inferior vena cava segment with thrombosis was extracted below the ligation point.The mice in the control group were not ligated,and the inferior vena cava segment at the same position as the experimental group was extracted.The expression changes of IL-10 and TGF-β1 were detected by immunohistochemistry(IHC),Western blotting and real-time qPCR.Results IHC results revealed that IL-10 was mainly expressed in monocytes in thrombosis and TGF-β1 was mainly expressed in monocytes and fibroblast-like cells in thrombosis.Western blotting and real-time qPCR showed that the relative expression levels of IL-10 and TGF-β1 in each experimental group were higher than those in the control group.The mRNA and protein levels of IL-10 reached the peak at 7 d and 10 d after ligation,respectively.The mRNA expression level at 7 d after ligation was 4.72±0.15 times that of the control group,and the protein expression level at 10 d after ligation was 7.15±0.28 times that of the control group.The mRNA and protein levels of TGF-β1 reached the peak at 10 d and 14 d after ligation,respectively.The mRNA expression level at 10 d after ligation was 2.58±0.14 times that of the control group,and the protein expression level at 14 d after ligation was 4.34±0.19 times that of the control group.Conclusion The expressions of IL-10 and TGF-β1 during the evolution of deep vein thrombosis present time-dependent sequential changes,and the expression levels of IL-10 and TGF-β1 can provide a reference basis for thrombus age estimation.

关 键 词：法医病理学 深静脉血栓形成 白细胞介素-10 转化生长因子-Β1 时序性表达 小鼠 

分 类 号：R89[医药卫生—法医学] DF795.1[医药卫生—临床医学] D919.1[政治法律—诉讼法学]