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Journal of Central South University Medical Sciences logoLink to Journal of Central South University Medical Sciences
. 2023 Nov 28;48(11):1650–1658. [Article in Chinese] doi: 10.11817/j.issn.1672-7347.2023.230285

经皮冠状动脉介入治疗术后支架内再狭窄患者血浆Rap1A水平变化及其机制（英文）

Changes of plasma Rap1A levels in patients with in-stent restenosis after percutaneous coronary intervention and the underlying mechanisms

GAO Haodong 1,2, KUANG Yuanyuan 1, LIU Yubo 1, ZHANG Yinzhuang 2, WANG Ping 1, MA Qilin 1,✉
Editor: 陈 丽文
PMCID: PMC10929945  PMID: 38432855

Abstract

Objective

Percutaneous coronary intervention (PCI) is one of the most important treatments for coronary artery disease (CAD). However, in-stent restenosis (ISR) after PCI is a serious complication without effective measures for prevention and treatment. This study aims to investigate the Ras-related protein 1A (Rap1A) level in ISR patients and in the tumor necrosis factor-α (TNF-α)-induced inflammatory injury model of human umbilical vein endothelial cells (HUVECs), to explore the role of Rap1A in regulating TNF-α-induced inflammation in HUVECs and to provide a new potential target for ISR prevention and treatment.

Methods

A total of 60 CAD patients, who underwent PCI between December 2020 and July 2022 from the Department of Cardiovascular Medicine of Xiangya Hospital, Central South University, and re-examined coronary angiography (CAG) 1 year after the operation, were included. After admission, 27 patients were diagnosed with ISR and 33 patients were diagnosed with non-in-stent restenosis (non-ISR) according to the CAG. Clinical data were collected, and the plasma Rap1A level was determined by enzyme linked immunosorbent assay (ELISA). In cell experiments, an inflammatory injury model was established with TNF-α treatment (10 ng/mL, 24 h) in HUVECs. The mRNA and protein expression levels of Rap1A, interlukin-6 (IL-6), and vascular cell adhesion molecule-1 (VCAM-1) were measured by real-time reverse transcription PCR and Western blotting. Small interfering RNA (siRNA) was used to explore the role of Rap1A in regulating TNF-α-induced inflammation in HUVECs.

Results

Compared with the non-ISR patients, a higher proportion of ISR patients had a history of smoking (P=0.005) and diabetes (P=0.028), and higher levels of glycosylated hemoglobin (HbA1c) (P=0.012), low-density lipoprotein cholesterol (LDL-c) (P=0.014), and hypersensitive C-reactive protein (hs-CRP) (P=0.027). The remaining projects did not show significant differences (all P>0.05). The plasma level of Rap1A in the ISR group was significantly higher than that in the non-ISR group [942.14 (873.28 to 1 133.81) μg/mL vs 886.93 (812.61 to 930.98) μg/mL; P=0.004]. Diabetes, LDL-c, and Rap1A were risk factors for ISR by univariate logistic regression analysis (all P<0.05). The mRNA and protein expression levels of inflammatory factors IL-6 and VCAM-1 were increased in HUVECs after 10 ng/mL TNF-α treatment for 24 h compared with the control group (all P<0.05), while the mRNA and protein levels of Rap1A were increased (both P<0.05). After inhibition of Rap1A in HUVECs, the mRNA and protein expression levels of IL-6 and VCAM-1 were significantly decreased (all P<0.05).

Conclusion

The plasma Rap1A level was significantly elevated in patients with ISR, suggesting that Rap1A may be a potential biomarker for predicting ISR. In the TNF-α- induced HUVECs inflammatory injury model, the expression level of Rap1A was increased. The level of TNF-α-induced endothelial cell inflammation was decreased after inhibition of Rap1A expression, suggesting that Rap1A may be a potential target for the treatment of endothelial cell inflammation in ISR.

Keywords: in-stent restenosis, Ras-related protein 1A, endothelial cell, inflammatory injury

冠状动脉疾病(coronary artery disease，CAD)严重威胁人类生命健康，1990至2017年间，全球增加的冠心病死亡病例中，中国约占38.2%[1]。经皮冠状动脉介入治疗(percutaneous coronary intervention，PCI)是CAD最有效的治疗方法之一，可改善患者的预后[2]。目前，PCI术后支架内再狭窄(in-stent restenosis，ISR)是影响冠心病行PCI术患者长期预后的重要因素，其患病率约占所有冠心病行PCI支架植入术患者的10%，带来严重的医疗负担[3-5]。因此，明确ISR的危险因素及发生机制，并进行早期预测及干预具有重要的临床意义。

PCI术后6个月至1年内发生的血管内膜新生是 ISR进展的主要机制之一。血管内皮细胞是构成血管内膜的重要组成部分，其主要病理生理机制包括介导炎症反应、调节血管张力、抗凝促纤溶、调节体液稳态及屏障功能等，其中介导炎症反应十分重要[6-7]。炎症反应是动脉粥样硬化及PCI术后ISR的重要病理过程，血管内皮细胞功能紊乱是引起动脉粥样硬化及冠心病PCI术后ISR的启动因素[8-9]。Ras相关蛋白1A(Ras-related protein 1A，Rap1A)是小分子G蛋白Ras超家族的成员，定位于细胞膜、细胞质及核周区域，在血管、心室、胃肠黏膜、淋巴结等部位广泛表达，具有多种生物学功能[10]。Rap1A是内皮细胞整合素活化所必需的分子，参与调节整合素介导的白细胞、血小板、卵巢癌细胞等各种细胞的黏附作用[11]。Rap1A通过黏附于肿瘤内皮细胞，促进髓系细胞招募并浸润到肿瘤微环境，参与肿瘤炎症反应及血管生成[12]，而血管内皮细胞炎症损伤在ISR的发生、发展中有至关重要的作用。目前Rap1A与PCI术后ISR的关系及其在血管内皮细胞中的作用机制鲜见报道。因此，本研究拟通过比较ISR患者与non-ISR患者血浆中Rap1A水平，探索两者之间的相关性。同时，通过构建TNF-α诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells，HUVECs)炎症损伤模型来探索Rap1A是否介导内皮细胞炎症。

1. 对象与方法

1.1. 对象

收集2020年12月至2022年7月因冠心病于中南大学湘雅医院心血管内科住院，且既往行PCI支架植入术的患者。纳入标准：1)根据《稳定性冠心病基层诊疗指南(2020年)》[13]，符合冠心病的诊断标准，并且行PCI术植入药物洗脱支架，术后1年复查冠状动脉造影(coronary arteriography，CAG)的患者；2)术后1年口服阿司匹林联合氯吡格雷或替格瑞洛行双联抗血小板和他汀类药物行调脂治疗的患者。排除标准：1)根据《稳定性冠心病基层诊疗指南(2020年》，应选择冠状动脉搭桥手术(coronary artery bypass graft，CABG)，但因患者坚持要求而行PCI的患者；2)首次PCI后30 d内发生急性心肌梗死、支架血管急性或亚急性闭塞等主要负性心血管事件的患者；3)植入支架发生断裂者；4)合并其他心血管疾病，如扩张型心肌病、心脏瓣膜病、先天性心脏病等的患者；5)合并肝肾功能不全、恶性肿瘤、风湿免疫性疾病、甲状腺疾病、妊娠或其他影响炎症水平的疾病如肺部感染或泌尿系感染等的患者。本研究采取ISR的影像学定义，即随访期间CAG冠状动脉支架内全程和/或支架两端5 mm以内的血管直径狭窄率≥50%的病变。本研究通过中南大学湘雅医院临床医学伦理委员会审核(审批号：2021101108)，患者均签署知情同意书。

1.2. 标本收集及处理

记录所有研究对象于入院后的一般资料，包括人口学资料(年龄、性别)、合并症(高血压、糖尿病)、生活方式(吸烟、饮酒)、体格检查(心率、收缩压和舒张压)等。于术前清晨采集患者空腹外周静脉血送医院检验科，采用全自动生化仪测定高密度脂蛋白胆固醇(high-density lipoprotein cholesterol，HDL-c)、胆固醇(total cholesterol，TC)、三酰甘油(triglyceride，TG)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol，LDL-c)、谷草转氨酶、肌酸激酶同工酶、糖化血红蛋白(glycosylated hemoglobin，HbA1c)、肌酐、尿酸、超敏C反应蛋白(high sensitive C-reactive protein，hs-CRP)。于医院心脏彩超室完善心脏彩色多普勒超声检查并记录左室射血分数(left ventricular ejection fraction，LVEF)。CAG结果由2名资深心血管介入医师判读。

患者入院24 h内，从其肘静脉采集空腹外周血3 mL置于EDTAK2抗凝采血管中，1 h内以3 000 r/min于4 ℃离心机内进行离心，时间为10 min，用移液枪小心收集上清液体并分装至离心管内，标记好信息，放入-80 ℃冰箱保存备用。使用及转移标本时，避免反复冻融。应用酶联免疫吸附试验 (enzyme linked immunosorbent assay，ELISA)(上海凡科维有限公司产品)检测血浆Rap1A水平。

1.3. 细胞培养及转染

HUVECs在含10% 胎牛血清的杜氏改良Eagle 培养基/营养混合物 F-12(Dulbecco’s modified eagle medium/nutrient mixture F-12，DMEM F-12)培养基，在37 ℃、5% CO2条件下培养。待细胞融合至50%~70%，使用10 ng/mL TNF-α(美国PeproTech公司产品)刺激24 h构建HUVECs炎症损伤模型。使用riboFECTTM CP转染试剂盒和siRNA(广州锐博生物技术有限公司)制备50 nmol/L的转染复合物，待细胞融合至50%~70%加入转染复合物处理72 h。Rap1A siRNA序列为5'-GCCAGAATTTAGCAAGACA-3'。使用蛋白质印迹法和实时反转录聚合酶链反应(real-time reverse transcription PCR，real-time RT-PCR)评估基因的敲除效率。siRNA NC为阴性对照。

1.4. RT-PCR

使用TRIzol试剂分离出细胞总RNA，cDNA反转录试剂盒(南京诺唯赞生物科技股份有限公司产品)反转录为cDNA。使用QuantStudio RT-PCR系统软件检测mRNA水平，通过2-ΔΔCt方法计算mRNA相对表达水平，β-actin作为内参，引物序列具体见表1。

表1.

引物序列

Table 1 Primer sequences

Gene names Forward Reverse
β-actin 5'-ATGTACGTTGCTATCCAGGC-3' 5'-CTCCTTAATGTCACGCACGAT-3'
Rap1A 5'-TCAGTCCACGTTTAACGACTTA-3' 5'-AGTTACACCACTGTCTTGCTAA-3'
IL-6 5'-CACTGGTCTTTTGGAGTTTGAG-3' 5'-GGACTTTTGTACTCATCTGCAC-3'
VCAM-1 5'-CAGGCTGGAGATAGACTTACTG-3' 5'-CCTCAATGACAGGAGTAAAGGT-3'
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Rap1A: Ras-related protein 1A; IL-6: Interleukin-6; VCAM-1: Vascular cell adhesion molecule-1.

1.5. 蛋白质印迹法

使用细胞裂解液提取细胞总蛋白，BCA蛋白检测试剂盒(上海碧云天生物技术有限公司产品)测定蛋白质浓度。将等量蛋白质进行电泳，随后转移到聚偏二氟乙烯(polyvinylidenefluoride，PVDF)膜上，将膜放入封闭液中进行封闭；分别加入Rap1A(武汉爱博泰克生物科技有限公司产品)、白细胞介素-6(interleukin-6，IL-6；成都正能生物技术有限责任公司产品)、血管细胞黏附分子-1(vascular cell adhesion molecule-1，VCAM-1；成都正能生物技术有限责任公司产品)、甘油醛-3-磷酸脱氢酶(glyceraldehyde-3-phosphate dehydrogenase，GAPDH)(美国PeproTech公司产品)一抗孵育过夜，洗膜后加入二抗孵育。加入显影剂后在化学发光成像系统中采集图像，使用Image J进行灰度分析。

1.6. 统计学处理

采用SPSS 21.0统计学软件进行数据分析。计数资料采用例数(%)进行描述，使用χ 2检验进行组间分布差异分析。正态分布的计量资料用均数±标准差( x¯ ±s)进行描述，并采用两独立样本t检验进行比较。非正态分布的计量资料用中位数(第1四分位数，第3四分位数)[M(P 25, P 75)]进行描述，并采用两独立样本的秩和检验进行比较。采用单因素logistic回归分析ISR的影响因素。在细胞实验中，采用双侧t检验来评估2组间差异；多组间比较采用单因素方差分析。P<0.05为差异具有统计学意义。

2. 结 果

2.1. 患者基线资料分析

本研究共纳入60例既往接受PCI治疗且1年后复查CAG的CAD患者，研究队列的基线资料包括临床资料和实验室指标(表2)。与non-ISR患者相比，ISR患者中有吸烟史(P=0.005)及合并糖尿病(P=0.028)者比例更高，HbA1c(P=0.012)、LDL-c(P=0.014)及hs-CRP(P=0.027)水平升高，余指标在2组间的差异无统计学意义(均P>0.05)。

表2.

ISR组与non-ISR组一般临床资料对比

Table 2 Comparison of general data of patients between the ISR group and the non-ISR group

Groups n Male/[No.(%)] Age/year

Smoking/

[No.(%)]

Diabetes/

[No.(%)]

 Systolic blood pressure*/mmHg Diastolic blood pressure*/mmHg
ISR 27 17(62.96) 59.78±9.13 16(59.26) 21(77.78) 129.37±18.02 79.00±8.86
non-ISR 33 24(72.73) 65.97±9.70 3(9.09) 16(48.48) 130.76±20.78 75.18±11.88
χ 2/t/U 0.654 0.118 17.272 4.832 0.860 1.621
P 0.317 0.906 0.005 0.028 0.393 0.110
Groups Uric acid†/(μmol·L-1) Creatinine†/(μmol·L-1) HbA1c*/% CK-MB*/(U·L-1) hs-CRP†/(mg·L-1)
ISR 346.7(299.7, 403.5) 81.00(65.43, 93.93) 7.21±1.42 16.39±5.88 2.50(1.35, 3.25)
non-ISR 352.5(300.05, 406.4) 83.00(69.5, 91.28) 6.48±0.97 15.06±5.61 1.90(1.18, 2.29)
χ 2/t/U 447.000 212.000 2.592 0.429 594.500
P 0.982 0.599 0.012 0.669 0.027
Groups Total cholesterol*/(mmo·L-1) HDL-c*/(mmol·L-1) LDL-c*/(mmol·L-1) Triglyceride*/(mmol·L-1) LVEF*/%
ISR 4.22±1.45 0.97±0.23 3.29±1.46 1.64±0.76 49.54±16.76
non-ISR 3.16±0.75 1.04±0.25 2.12±0.64 1.84±1.55 57.64±12.10
χ 2/t/U 1.853 0.385 2.519 0.693 1.337
P 0.069 0.702 0.014 0.491 0.186
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*Data are expressed as mean±standard deviation ( x¯ ±s); †Data are expressed as M(P 25, P 75). 1 mmHg=0.133 kPa. ISR: In-stent restenosis; non-ISR: Non in-stent restenosis; HbA1c: Glycosylated hemoglobin; HDL-c: High-density lipoprotein cholesterol; LDL-c: Low-density lipoprotein cholesterol; hs-CRP: Hypersensitive C-reactive protein; CK-MB: Creatine kinase isoenzyme; LVEF: Left ventricular ejection fraction; n: number of cases.

2.2. ISR患者血浆Rap1A表达水平变化

ELISA法结果显示：ISR组的血浆Rap1A水平显著高于non-ISR组[942.14(873.28~1 133.81) μg/mL vs 886.93(812.61~930.98) μg/mL，P=0.004；图1]。

图1.

图1

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ISR组与non-ISR组血浆Rap1A散点图对比

Figure 1 Comparison of plasma Rap1A scatter plots between the ISR group and the non-ISR group

**P<0.01. ISR: In-stent restenosis; non-ISR: Non-in-stent restenosis.

2.3. 单因素logistic回归分析

单因素logistic回归分析显示：糖尿病、LDL-c、Rap1A均是ISR的危险因素(均P<0.05，表3)

表3.

单因素logistic回归分析

Table 3 Univariate logistic regression analysis

Variables OR (95% CI) P
Diabetes 3.50(1.12-10.96) 0.031
LDL-c/(mmol·L-1) 4.64(1.92-11.25) 0.001
Ln-transformed hs-CRP/(mg·L-1) 3.58(1.16-11.02) 0.026
Rap1A
T1 (<865.61 μg/mL) Reference
T2 (865.62-942.14 μg/mL) 1.56(0.44-5.76) 0.508
T3 (≥942.15 μg/mL) 4.33(1.15-16.32) 0.030
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Rap1A was converted to the grade variables T1, T2, and T3 according to the interquartile method. LDL-c: Low-density lipoprotein cholesterol; ln-transformed: Log-transformed; hs-CRP: Hypersensitive C-reactive protein.

2.4. Rap1A在HUVECs炎症损伤模型中的作用

2.4.1. Rap1A在TNF-α诱导的HUVECs炎症损伤模型中的表达

与对照组相比，10 ng/mL TNF-α诱导24 h后，HUVECs中炎症因子IL-6、VCAM-1的mRNA及蛋白质表达水平升高，同时Rap1A mRNA及蛋白质水平升高(均P<0.05，图2)。

图2.

图2

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Rap1A在TNF-α诱导的HUVECs炎症损伤模型中表达

Figure 2 Rap1A expression in TNF-α-induced inflammatory injury model of HUVECs

A: Rap1A, IL-6, and VCAM-1 protein expression levels after 10 ng/mL TNF-α treatment of HUVECs for 24 h; B: Rap1A, IL-6, and VCAM-1 mRNA expression levels after 10 ng/mL TNF-α treatment of HUVECs for 24 h. Data are expressed as mean±standard deviation. *P<0.05; **P<0.01; ***P<0.001; n=3. Rap1A: Ras-related protein 1A; IL-6: Interleukin-6; VCAM-1: Vascular cell adhesion molecule-1; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; HUVECs: Human umbilical vein endothelial cells; TNF-α: Tumor necrosis factor-α.

2.4.2. Rap1A siRNA转染效率验证

RT-PCR和蛋白质印迹法结果显示：与对照组及siNC组相比，siRNA组Rap1A mRNA及蛋白质表达水平均显著降低(均P<0.05，图3)。

图3.

图3

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Rap1A siRNA转染HUVECs后Rap1A蛋白及mRNA表达水平

Figure 3 Rap1A protein and mRNA expression after Rap1A siRNA transfection of HUVECs

Expression of Rap1A protein level (A) and mRNA level (B) after transfection with 50 nmol/L Rap1A siRNA in HUVECs for 48 h. Data are expressed as mean±standard deviation. **P<0.01, ***P<0.001; n=3. Rap1A: Ras-related protein 1A; HUVECs: Human umbilical vein endothelial cells; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.

2.4.3. 干扰Rap1A后TNF-α诱导的HUVECs炎症反应水平

在敲低Rap1A后，TNF-α诱导的IL-6、VCAM-1的mRNA及蛋白质表达水平均显著降低(均P<0.05，图4)。

图4.

图4

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Rap1A siRNA转染HUVECs后IL-6、VCAM-1蛋白质及mRNA表达水平

Figure 4 IL-6, VCAM-1 protein and mRNA expression after Rap1A siRNA transfection in HUVECs

IL-6, VCAM-1 protein and mRNA expression after transfection with 50 nmol/L Rap1A siRNA for 48 h and treatment with 10 ng/mL TNF-α for 24 h in HUVECs. Data are expressed as mean±standard deviation. *P<0.05; **P<0.01; n=3. Rap1A: Ras-related protein 1A; HUVECs: Human umbilical vein endothelial cells; IL-6: Interleukin-6; VCAM-1: Vascular cell adhesion molecule-1; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; TNF-α: Tumor necrosis factor-α.

3. 讨 论

随着冠心病患病率逐年增加，PCI手术量也随之上升，支架植入后发生的ISR始终是困扰手术医生及患者的常见并发症之一。ISR的发生可能与血管内皮细胞功能紊乱和新生动脉粥样硬化有关[14]，但其具体机制尚不明确。因此，目前仍缺乏针对PCI术后ISR的有效防治措施。现有关于冠心病植入DES药物洗脱支架(drug eluting stent，DES)后ISR发生及预测因素分析的临床研究[15]发现，与ISR有关的危险因素包括病变血管因素、患者因素及不可控的手术因素等。在这些因素中，患者因素主要包括合并代谢紊乱或存在炎症反应。支架是一种强致炎因子，在支架植入后可观察到炎症细胞聚集浸润[16]。当血管内皮损伤后，血液循环中的单核细胞被募集到冠脉损伤部位，造成纤维蛋白沉积、血小板活化，导致血栓形成，引起ISR；此外，炎症反应和血栓形成具有共同的信号转导途径，并且炎症反应促进凝血级联反应的激活、刺激血小板活化、引起血管内皮细胞功能紊乱和细胞外基质沉积，最终导致DES部位动脉延迟愈合、内皮细胞剥脱、新生内膜增厚和ISR的发生[14, 17-18]。本研究通过比较ISR组与non-ISR组患者的基线资料，发现与non-ISR组对比，糖尿病、HbA1c、LDL-c及hs-CRP在ISR组升高，这与既往研究[19]基本相符。炎症反应在ISR的发生、发展中起重要作用。本研究进一步通过单因素logistic回归分析发现，hs-CRP是ISR的危险因素。CRP由肝细胞合成，可参与人体免疫反应，在既往研究[20-21]中发现其可作为预测心脑血管事件的炎症标志物。

血管内皮细胞炎症损伤在ISR的发生、发展中起重要作用。TNF-α或其他炎症细胞因子可刺激血管内皮细胞，促进其分泌促炎因子(如IL-1、IL-6等)及细胞黏附分子(如VCAM-1、ICAM-1等)[22]。IL-6可以放大炎症反应，与微血管功能受损和心血管疾病的进展有关[23]。VCAM-1可以通过介导白细胞通过血管壁进入内皮，在内皮细胞炎症损伤中起关键作用[24]。Rap1A在细胞膜与细胞核之间起信号传递作用，可与RAF原癌基因丝苏氨酸蛋白激酶(RAF proto-oncogene serine/threonine-protein kinase，Raf-1)、Ral鸟嘌呤核苷酸解离刺激因子(Ral guanine nucleotide dissociation stimulator，RalGDs)等结合[25]。Rap1A通过黏附于肿瘤内皮细胞，促进髓系细胞招募并浸润到肿瘤微环境，参与肿瘤炎症反应及血管生成[12]。但目前Rap1A与血管内皮细胞之间的关系未见报道。本研究首次发现Rap1A在ISR组升高。进一步通过单因素logistic回归分析发现，Rap1A是ISR的危险因素。通过构建TNF-α诱导的HUVECs炎症损伤模型，发现在正常HUVECs中加入TNF-α(10 ng/mL)后炎症标志物IL-6、VCAM-1升高，同时Rap1A也明显升高，进一步通过干扰TNF-α诱导的HUVECs炎症损伤模型中的Rap1A表达，发现IL-6、VCAM-1随之低表达。这提示Rap1A可能参与HUVECs炎症反应的发生。其机制可能为Rap1A是内皮细胞整合素活化所必需的因子。首先，香叶基转移酶进行异戊烯化来翻译后修饰Rap1A并使其激活[26]。其次，Rap1Ac端含有丰富的正电荷残基(赖氨酸和精氨酸)，这些残基有利于募集到带负电荷且富含磷脂酰肌醇-4,5-二磷酸(phosphatidylinositol 4,5-bisphosphate，PIP2)和磷脂酰肌醇-3,4,5-三磷酸(phosphatidylinositol 3,4,5-trisphosphate，PIP3)等磷脂的细胞膜区，这些磷脂很可能通过调控由Rap1A等组成的整合素激活复合物的组装，参与调节整合素介导的炎症细胞、血小板等黏附过程[27-28]。整合素分布于内皮细胞及白细胞表面，能够介导白细胞与内皮细胞的黏附以及白细胞向内皮下迁移[29]。白细胞黏附于血管内皮，形成炎症反应，引起血管内皮细胞功能紊乱，从而导致ISR的发生[30-31]。因此，Rap1A可能是ISR中血管内皮功能紊乱的一个新的生物标志物及防治靶点。

本研究存在以下局限性：1)为单中心研究，样本例数偏少，故仅分析ISR与Rap1A相关性，后续将进一步扩大样本量，同时完善预后分析。2)仅从细胞水平分析Rap1A促进炎症的作用机制，未通过动物实验进行验证，后续将进一步完善。Rap1A与冠心病PCI术后ISR之间的关系需进一步深入研究。在ISR的发生、发展中，白细胞和其他类型的细胞也参与了内皮细胞功能紊乱的病理生理过程，可作为未来研究的重点。

综上所述，在冠心病PCI术后ISR患者中血浆Rap1A蛋白表达水平升高，是ISR的危险因素，提示Rap1A可能是ISR的潜在生物标志物。在TNF-α诱导的内皮细胞炎症损伤模型中，Rap1A的表达水平增高，而使用siRNA敲减内皮细胞Rap1A基因表达后，TNF-α诱导的内皮细胞炎症反应水平下降，提示Rap1A可能是治疗ISR内皮细胞炎症的潜在靶点。

基金资助

国家自然科学基金(81974026)；湖南省自然科学基金(2022JJ30069)。

This work was supported by the National Natural Science Foundation (81974026) and the Natural Science Foundation of Hunan Province (2022JJ30069), China.

利益冲突声明

作者声称无任何利益冲突。

作者贡献

高浩东 研究设计，数据分析，论文撰写与修改；匡圆圆、刘昱博、张银妆、王萍 数据分析、论文修改；马琦琳 研究设计，论文修改。所有作者阅读并同意最终的文本。

Footnotes

http://dx.chinadoi.cn/10.11817/j.issn.1672-7347.2023.230285

原文网址

http://xbyxb.csu.edu.cn/xbwk/fileup/PDF/2023111650.pdf

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