脓毒症大鼠模型肝素结合蛋白含量变化与凝血功能紊乱和血管内皮损伤的关系

doi:10.12300/j.issn.1674-5817.2025.182

摘要/Abstract

摘要：

目的通过观察脓毒症状态下SD大鼠肝素结合蛋白（heparin-binding protein，HBP）、凝血功能指标及血管内皮损伤标志物的动态变化，分析各指标间的关联特征，阐明HBP在脓毒症相关性凝血紊乱中的调控作用。方法取18只8～12周龄SPF级雄性SD大鼠，采用盲肠结扎穿孔术建立脓毒症模型，于建模前（0 h）及建模后3 h、6 h、9 h进行心脏采血，利用血栓弹力图（thromboelastography，TEG）技术（含肝素酶杯）检测普通杯和肝素酶杯凝血反应时间（R值）之差（ΔR），以评估凝血功能及内源性肝素效应。采用ELISA法测定血浆中HBP、硫酸乙酰肝素（heparan sulfate，HS）、多配体蛋白聚糖-1（syndecan-1，SDC-1）及血栓调节蛋白（thrombomodulin，TM）水平。将ΔR≥2 min的标本归为ΔR≥2 min组，将ΔR<2 min的标本归为ΔR<2 min组。运用相关性和接受者操作特征（receiver operating characteristic，ROC）曲线分析、评估HBP浓度与各指标间的关系及其对ΔR值的预测效能。结果与0 h相比，ΔR、HS、SDC-1和HBP水平在建模后显著升高（P<0.01）。ΔR≥2 min组大鼠的HBP、SDC-1和HS水平显著高于ΔR<2 min组（P<0.05或P<0.01）。HBP与ΔR、HS呈显著正相关（P<0.05）；ΔR与HS呈显著正相关（r=0.799，P<0.01），与SDC-1呈弱正相关（r=0.289，P<0.05）。HS、HBP和SDC-1对ΔR≥2 min具有显著预测价值（P<0.01）。结论脓毒症早期，糖萼降解释放HS类抗凝物质可引起凝血紊乱，而HBP可反映该病理进程，在脓毒症相关凝血紊乱的发生发展中发挥重要调控作用。

关键词: 脓毒症, 肝素结合蛋白, 硫酸乙酰肝素, 血栓弹力图, 内皮损伤, SD大鼠

Abstract:

Objective To observe the dynamic changes in heparin-binding protein (HBP), coagulation function indicators, and vascular endothelial injury markers in SD rats with sepsis, analyze the associations among these indicators, and elucidate the regulatory role of HBP in sepsis-associated coagulation disorders. Methods Eighteen specific pathogen-free (SPF) male SD rats aged 8–12 weeks were selected to establish a sepsis model using cecal ligation and puncture (CLP). Blood samples were collected from the heart at 0 hours before modeling and at 3, 6, and 9 hours post-modeling. Thromboelastography (TEG) with standard and heparinase cups was used to determine the difference in reaction time (R value) between the two cups (ΔR), thereby evaluating coagulation function and the endogenous heparin effect. Plasma levels of HBP, heparan sulfate (HS), syndecan-1 (SDC-1), and thrombomodulin (TM) were measured by enzyme-linked immunosorbent assay (ELISA). Samples with ΔR≥2 min were grouped as the ΔR≥2 min group, while those with ΔR<2 min were grouped as the ΔR<2 min group. Correlation and receiver operating characteristic (ROC) curve analyses were performed to evaluate the relationship between HBP and various indicators, as well as its predictive efficacy for ΔR values. Results ΔR, HS, SDC-1 and HBP significantly increased after modeling compared with 0 hours (P<0.01). The HS, SDC-1, and HBP levels in the ΔR≥2 min group were significantly higher than those in the ΔR<2 min group (P<0.05 or P<0.01). HBP showed a significant positive correlation with ΔR and HS (P<0.05), while ΔR was significantly positively correlated with HS (r=0.799, P<0.01) and weakly positively correlated with SDC-1 (r=0.289, P<0.05). HS, HBP, and SDC-1 demonstrated significant predictive value for ΔR≥2 min (P<0.01). Conclusion In the early stages of sepsis, the release of HS-like anticoagulant substances from the glycocalyx following its degradation can lead to coagulation disorders, while HBP can reflect the pathological process and play an important regulatory role in the occurrence and development of sepsis-related coagulation disorders.

Key words: Sepsis, Heparin-binding protein, Heparan sulfate, Thromboelastography, Endothelial injury, SD rats

中图分类号:

R631
R-332

田艳,赖冬,赖飞,等. 脓毒症大鼠模型肝素结合蛋白含量变化与凝血功能紊乱和血管内皮损伤的关系[J]. 实验动物与比较医学, 2026, 46(5): 700-706. DOI: 10.12300/j.issn.1674-5817.2025.182.

TIAN Yan,LAI Dong,LAI Fei,et al. Study on the Relationship Between Changes in Heparin-Binding Protein Levels and Coagulation Disorders and Vascular Endothelial Injury in a Rat Model of Sepsis[J]. Laboratory Animal and Comparative Medicine, 2026, 46(5): 700-706. DOI: 10.12300/j.issn.1674-5817.2025.182.

图/表 4

表1

建模后不同时间点大鼠HS、TM、SDC-1、ΔR与HBP的变化

Table 1

Changes of HS, TM, SDC-1, ΔR，and HBP of rats at different time points after modeling

指标 Index	建模后时间/h Post-modeling time/h
指标 Index	0	3	6	9
HS ρ/(ng·mL^-1)	6.67 (5.60, 11.26)	6.74 (5.58, 10.01)	15.62 (10.63, 21.56)^**	14.43 (8.37, 18.43)^**
TM ρ/(ng·mL^-1)	6.13 (4.43, 10.58)	6.18 (2.31, 10.19)	7.54 (4.46, 10.87)	5.02 (2.04, 6.33)
SDC-1 ρ/(ng·mL^-1)	14.42 (11.48, 18.99)	16.34 (12.13, 19.24)	23.89 (18.01, 26.70)^**	21.88 (15.10, 25.77)^**
ΔR t/min	0.35 (0.08, 0.90)	0.65 (0.38, 3.25)^*	3.55 (2.45, 5.00)^**	3.00 (2.00, 3.70)^**
HBP ρ/（pg·mL^-1）	152 (115, 226)	326 (262, 417)^**	387 (263, 493)^**	342 (222, 486)^**

表2

ΔR≥2 min组与ΔR<2 min组大鼠血浆中HS、TM、SDC-1、ΔR与HBP的ELISA检测结果

Table 2

ELISA results of HS, TM, SDC-1, ΔR， and HBP in the plasma of rats in the ΔR≥2 min group and the ΔR<2 min group

指标 Index	ΔR≥2 min组/n=26 ΔR≥2 min group/n=26	ΔR＜2 min组/n=46 ΔR＜2 min group/n=46	z	P
HS ρ/(ng·mL^-1)	18.43（5.73, 21.99)	8.07（5.75, 15.87)	-2.537	0.011
TM ρ/(ng·mL^-1)	6.23（4.08, 10.19)	5.70（2.04, 6.45)	-1.559	0.119
SDC-1 ρ/(ng·mL^-1)	25.88（23.10, 27.77)	16.73（13.11, 22.63)	-4.628	0.000
ΔR t/min	3.00（2.18, 3.70)	0.00（-0.23, 0.53)	-3.936	0.000
HBP ρ/（pg·mL^-1）	363（322，498）	122（111，204）	-6.875	0.000

图1 TM、SDC-1、HBP、ΔR与HS指标间的相关性分析结果Figure 1 Results of correlation analysis between TM, SDC-1, HBP, ΔR and HS

注：TM，血栓调节蛋白；SDC‑1，多配体聚糖-1；HBP，肝素结合蛋白；ΔR，凝血反应时间普通杯与肝素酶杯R值之差；HS，硫酸乙酰肝素。*P<0.05，**P<0.01。

Note： TM， thrombomodulin； SDC‑1， syndecan-1； HBP， heparin-binding protein； ΔR， difference in clotting reaction time （R value） between standard and heparinase cups； HS， heparan sulfate. *P<0.05， **P<0.01.

图2 HS、HBP和SDC-1预测ΔR≥2 min的ROC曲线

注：ROC，相关性和接受者操作特征；HBP，肝素结合蛋白；HS，硫酸乙酰肝素；SDC‑1，多配体聚糖-1。

Figure 2 ROC curves for predicting Δ R ≥ 2 min using HS， HBP， and SDC-1

Note： ROC， receiver operating characteristic； HBP， heparin-binding protein； HS， heparan sulfate； SDC‑1， syndecan-1.

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