Juan-Juan Wu, Jun-Jie Huang, Yu Zhang, Jia-Ying Zhuo, Gang Chen, Shu-Han Yang, Yun-Qi Zhao, Yan-Yan Fan
{"title":"深静脉血栓小鼠体内 IL-10 和 TGF-β1 的时间依赖性序列变化","authors":"Juan-Juan Wu, Jun-Jie Huang, Yu Zhang, Jia-Ying Zhuo, Gang Chen, Shu-Han Yang, Yun-Qi Zhao, Yan-Yan Fan","doi":"10.12116/j.issn.1004-5619.2023.430506","DOIUrl":null,"url":null,"abstract":"<p><strong>Objectives: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>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.</p>","PeriodicalId":12317,"journal":{"name":"法医学杂志","volume":"40 2","pages":"179-185"},"PeriodicalIF":0.0000,"publicationDate":"2024-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Time-Dependent Sequential Changes of IL-10 and TGF-β1 in Mice with Deep Vein Thrombosis.\",\"authors\":\"Juan-Juan Wu, Jun-Jie Huang, Yu Zhang, Jia-Ying Zhuo, Gang Chen, Shu-Han Yang, Yun-Qi Zhao, Yan-Yan Fan\",\"doi\":\"10.12116/j.issn.1004-5619.2023.430506\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objectives: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>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.</p>\",\"PeriodicalId\":12317,\"journal\":{\"name\":\"法医学杂志\",\"volume\":\"40 2\",\"pages\":\"179-185\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-04-25\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"法医学杂志\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.12116/j.issn.1004-5619.2023.430506\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"Medicine\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"法医学杂志","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.12116/j.issn.1004-5619.2023.430506","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Medicine","Score":null,"Total":0}
引用次数: 0
摘要
研究目的检测小鼠深静脉血栓形成过程中白细胞介素-10（IL-10）和转化生长因子-β1（TGF-β1）的表达变化，探讨其在血栓年龄估计中的应用价值：方法：实验组小鼠均接受下腔静脉结扎手术。方法：实验组小鼠分别于结扎后 1 d、3 d、5 d、7 d、10 d、14 d 和 21 d 过度麻醉处死。在结扎点下方提取有血栓形成的下腔静脉段。对照组小鼠不结扎，提取与实验组相同位置的下腔静脉段。通过免疫组化（IHC）、Western 印迹和实时 qPCR 检测 IL-10 和 TGF-β1 的表达变化：结果：IHC 结果显示，血栓形成时 IL-10 主要在单核细胞中表达，TGF-β1 主要在单核细胞和成纤维细胞中表达。Western 印迹和实时 qPCR 显示，各实验组 IL-10 和 TGF-β1 的相对表达水平均高于对照组。IL-10 的 mRNA 和蛋白水平分别在结扎后 7 d 和 10 d 达到高峰。结扎后 7 d 的 mRNA 表达水平是对照组的 4.72±0.15 倍，结扎后 10 d 的蛋白表达水平是对照组的 7.15±0.28 倍。TGF-β1 的 mRNA 和蛋白水平分别在结扎后 10 d 和 14 d 达到峰值。结扎后 10 d 的 mRNA 表达水平是对照组的 2.58±0.14 倍，结扎后 14 d 的蛋白表达水平是对照组的 4.34±0.19 倍：结论：在深静脉血栓形成过程中，IL-10和TGF-β1的表达呈时间依赖性顺序变化，IL-10和TGF-β1的表达水平可为血栓年龄估计提供参考依据。
Time-Dependent Sequential Changes of IL-10 and TGF-β1 in Mice with Deep Vein Thrombosis.
Objectives: 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.
Conclusions: 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.
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