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

doi:10.12116/j.issn.1004-5619.2023.430506

Abstract

Abstract:

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.

Key words: forensic pathology, deep venous thrombosis, interleukin-10 （IL-10）, transforming growth factor-β1 （TGF-β1）, time-dependent sequential expression, mice

CLC Number:

Juan-juan WU, Jun-jie HUANG, Yu ZHANG, Jia-ying ZHUO, Gang CHEN, Shu-han YANG, Yun-qi ZHAO, Yan-yan FAN. Time-Dependent Sequential Changes of IL-10 and TGF-β1 in Mice with Deep Vein Thrombosis[J]. Journal of Forensic Medicine, 2024, 40(2): 179-185.

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References 29

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基因	引物序列（5′→3′）
GAPDH	F：AGGTCGGTGTGAACGGATTTG
	R：GGGGTCGTTGATGGCAACA
IL-10	F：GCTGTCATCGATTTCTCCCCT
	R：GACACCTTGGTCTTGGAGCTTAT
TGF-β1	F：ACTGGAGTTGTACGGCAGTG
	R：GGGGCTGATCCCGTTGATT

基因	引物序列（5′→3′）
GAPDH	F：AGGTCGGTGTGAACGGATTTG
	R：GGGGTCGTTGATGGCAACA
IL-10	F：GCTGTCATCGATTTCTCCCCT
	R：GACACCTTGGTCTTGGAGCTTAT
TGF-β1	F：ACTGGAGTTGTACGGCAGTG
	R：GGGGCTGATCCCGTTGATT

时间点	IL-10	TGF-β1
术后1 d	16.4±2.7	2.2±1.3
术后3 d	23.3±1.8	17.3±5.0^1）
术后5 d	64.3±2.5^1）	54.0±1.6^1）
术后7 d	73.0±3.9^1）	74.9±1.9^1）
术后10 d	81.2±2.8^1）	79.1±2.1
术后14 d	81.8±2.9	85.8±1.7^1）
术后21 d	68.8±1.4^1）	69.8±0.4^1）

时间点	IL-10	TGF-β1
术后1 d	16.4±2.7	2.2±1.3
术后3 d	23.3±1.8	17.3±5.0^1）
术后5 d	64.3±2.5^1）	54.0±1.6^1）
术后7 d	73.0±3.9^1）	74.9±1.9^1）
术后10 d	81.2±2.8^1）	79.1±2.1
术后14 d	81.8±2.9	85.8±1.7^1）
术后21 d	68.8±1.4^1）	69.8±0.4^1）

组别	IL-10		TGF-β1
组别	蛋白	mRNA	蛋白	mRNA
对照	1.00±0.00	1.00±0.08	1.00±0.00	1.00±0.06
1 d	3.16±0.20^1）	2.17±0.16^1）	1.79±0.06^1）	1.17±0.10
3 d	4.48±0.15^1）2）	3.18±0.22^1）2）	2.47±0.16^1）2）	1.79±0.07^1）2）
5 d	5.19±0.13^1）2）	3.32±0.19^1）	3.06±0.10^1）2）	2.00±0.08^1）
7 d	5.63±0.15^1）	4.72±0.15^1）2）	3.28±0.13^1）	2.28±0.09^1）
10 d	7.15±0.28^1）2）	3.31±0.15^1）2）	3.77±0.24^1）2）	2.58±0.14^1）2）
14 d	6.62±0.18^1）2）	2.19±0.06^1）2）	4.34±0.19^1）2）	2.02±0.12^1）2）
21 d	4.48±0.24^1）2）	0.53±0.04^1）2）	1.67±0.17^1）2）	1.14±0.10^2）