己糖胺生物合成通路在血管内皮炎症中的调控作用研究

doi:10.12114/j.issn.1007-9572.2024.0057

摘要/Abstract

摘要： 背景动脉粥样硬化（AS）是心血管病主要的病理基础，以血管内皮炎症为主要特征，因而靶向炎症相关机制是防治AS的关键。目的研究己糖胺生物合成通路（HBP）对黏附分子的影响及其在血管内皮炎症中的调控作用。方法 2022年8—12月，将24只SPF级C57BL/6雌性小鼠按体质量以随机区组设计方法分为对照组、6-重氮-5-氧代-L-正亮氨酸（DON）组、高脂饮食（HFD）组、HFD+DON组。各组小鼠给予相应干预措施15周后，收集小鼠的血清及主动脉组织。使用生化试剂盒检测干预前后小鼠的血脂水平，采用HE染色法检测主动脉根部的病理变化，并通过免疫荧光染色、ELISA和蛋白质印迹法检测细胞间黏附分子1（ICAM-1）和血管细胞黏附分子1（VCAM-1）的表达水平。结果干预15周后，与对照组相比，HFD组LDL-C、TC水平显著性上升，而HDL-C显著降低（P<0.05）；HFD组与HFD+DON组之间的血脂水平无变化（P>0.05）。HE染色结果显示，HFD组血管内膜增厚，血管平滑肌形态异常，结构紊乱，并可见大量的泡沫细胞。HFD+DON组的小鼠平滑肌细胞排列整齐，内皮细胞层连续，泡沫细胞数量明显减少，细胞间隙基本正常。免疫荧光染色、ELISA和蛋白质印迹法结果均显示，与HFD组相比，HFD+DON组的ICAM-1、VCAM-1蛋白表达下调。结论抑制HBP具有下调黏附分子ICAM-1、VCAM-1表达，改善血管内皮炎症的作用。

关键词: 血管内皮炎症, 己糖胺生物合成通路, 细胞间黏附分子1, 血管细胞黏附分子1, 6-重氮-5-氧代-L-正亮氨酸, 小鼠

Abstract:

Background

Atherosclerosis (AS) is the main pathological basis of cardiovascular disease and is characterized by vascular endothelial inflammation, thus targeting inflammation-related mechanisms is the key to prevention and treatment of AS.

Objective

To investigate the effect of the hexosamine biosynthesis pathway (HBP) on adhesion molecules and its regulatory role in vascular endothelial inflammation.

Methods

From August to December 2022, 24 SPF grade C57BL/6 female mice were divided into control group, DON group, HFD group, HFD+DON group according to randomized block design method using body weight stratification. Serum and aortic tissue from the mice were collected after 15 weeks of corresponding intervention measures in each group of mice. The lipid levels of mice were detected using biochemical kits before and after intervention, pathological changes in the aortic root were detected by HE staining, and the expression levels of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were detected by immunofluorescence staining, ELISA and Western blot.

Results

After 15 weeks of intervention, compared with the control group, the levels of LDL-C and TC were increased significantly in the HFD group, while HDL-C was reduced significantly (P<0.05) ; There was no change in the lipid levels between the HFD group and the HFD+DON group (P>0.05). HE staining results showed that the vascular intima was thickened, the morphology of vascular smooth muscle was abnormal, the structure was disorganized, and a large number of foam cells were seen in HFD group. The smooth muscle cells of mice were neatly aligned, the endothelial cell layer was continuous, the number of foam cells was reduced significantly, and the cell gap was normal basically in the HFD+DON group. The results of immunofluorescence staining, ELISA and Western blot showed that the expression of ICAM-1 and VCAM-1 was down-regulated in the HFD+DON group compared with the HFD group.

Conclusion

Inhibition of HBP can down-regulate the expression of ICAM-1 and VCAM-1, and play a role in improving vascular endothelial inflammation.

Key words: Vascular endothelial inflammation, Hexosamine biosynthetic pathway, Intercellular adhesion molecule-1, Vascular cell adhesion molecule-1, 6-diazo-5-oxo-L-norleucine, Mice

中图分类号:

R 543

陈伊静,许琪,刘忠典,等. 己糖胺生物合成通路在血管内皮炎症中的调控作用研究[J]. 中国全科医学, 2025, 28(15): 1871-1877. DOI: 10.12114/j.issn.1007-9572.2024.0057.
CHEN Yijing,XU Qi,LIU Zhongdian, et al. The Regulatory Role of Hexosamine Biosynthesis Pathway in Vascular Endothelial Inflammation[J]. Chinese General Practice, 2025, 28(15): 1871-1877. DOI: 10.12114/j.issn.1007-9572.2024.0057.

图/表 7

表1 4组小鼠血脂水平比较（±s，mmol/L）

Table 1 Comparison of blood lipid levels in 4 groups of mice

组别	只数	LDL-C				TC				HDL-C
组别	只数	干预前	干预15周后	t_配对值	P值	干预前	干预15周后	t_配对值	P值	干预前	干预15周后	t_配对值	P值
对照组	6	0.096±0.002	0.624±0.183	7.101	0.000 9	0.149±0.011	2.421±0.617	7.528	0.001 7	3.183±0.489	1.979±0.358	6.036	0.001 8
DON组	6	0.099±0.005	0.583±0.157	7.487	0.000 7	0.158±0.006	2.493±0.286	26.787	<0.001	2.593±0.820	1.812±0.351	2.131	0.100 1
HFD组	6	0.095±0.002	2.522±0.747^ab	7.208	0.000 4	0.149±0.015	5.149±1.838^ab	5.419	0.005 6	2.929±0.929	0.283±0.190^ab	5.220	0.006 4
HFD+DON组	6	0.098±0.004	2.541±0.646^ab	9.231	0.000 3	0.151±0.010	5.181±1.151^ab	11.597	0.000 3	3.038±0.327	0.387±0.429^ab	12.715	0.000 2
F值		1.530	28.780			0.620	11.370			0.570	43.50
P值		0.235	<0.001			0.609	<0.001			0.640	<0.001

图1 4组小鼠主动脉根部HE染色（×100）注：A为对照组，B为DON组，C为HFD组，D为HFD+DON组；C图中红色箭头表示主动脉管腔内存在泡沫细胞。

Figure 1 HE staining of aortic roots in 4 groups of mice

图2 4组小鼠主动脉根部横截面ICAM-1的表达（×200）注：A为对照组，B为DON组，C为HFD组，D为HFD+DON组；红色高亮为ICAM-1染色阳性。

Figure 2 Expression of ICAM-1 in cross-section of aortic root in 4 groups of mice

图3 4组小鼠主动脉根部横截面VCAM-1的表达（×200）注：A为对照组，B为DON组，C为HFD组，D为HFD+DON组；红色高亮为VCAM-1染色阳性。

Figure 3 Expression of VCAM-1 in cross-section of aortic root in 4 groups of mice

表2 4组小鼠血清黏附分子ICAM-1、VCAM-1水平比较（±s，pg/mL）

Table 2 Comparison of serum adhesion molecules ICAM-1 and VCAM-1 in 4 groups of mice

组别	只数	ICAM-1	VCAM-1
对照组	6	1 778.618±221.922	1 701.446±451.229
DON组	6	1 337.773±302.284	1 786.358±390.094
HFD组	6	2 003.233±763.840^b	2 269.460±427.058^a
HFD+DON组	6	826.039±241.942^abc	1 326.899±242.623^bc
F值		7.760	5.069
P值		0.003	0.011

表3 4组小鼠主动脉组织ICAM-1、VCAM-1蛋白表达比较（±s）

Table 3 Comparison of protein expression of ICAM-1 and VCAM-1 in aortic tissues of the 4 groups of mice

组别	只数	ICAM-1	VCAM-1
对照组	3	1.000±0	1.000±0
DON组	3	1.087±0.428	0.759±0.066
HFD组	3	1.243±0.272	1.325±0.181^ab
HFD+DON组	3	1.158±0.335	0.900±0.105^c
F值		0.58	19.30
P值		0.639	<0.001

图4 4组小鼠主动脉组织ICAM-1、VCAM-1蛋白表达情况注：A为对照组，B为DON组，C为HFD组，D为HFD+DON组；ICAM-1=细胞间黏附分子1，VCAM-1=血管细胞黏附分子1。

Figure 4 Protein expression of ICAM-1 and VCAM-1 in aortic tissues of the 4 groups of mice

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