Abstract：
BACKGROUND: Various factors, including blood, inflammatory, infectious, and immune factors, can cause ischemic stroke. However, the primary cause is often the instability of cervical arteriosclerosis plaque. It is estimated that 18-25% of ischemic strokes are caused by the rupture of carotid plaque.1 Plaque stability is crucial in determining patient prognosis. Developing a highly accurate, non-invasive, or minimally invasive technique to assess carotid plaque stability is crucial for diagnosing and treating stroke.Previous research by our group has demonstrated that the expression levels of CHOP (C/EBP homologous protein) and GRP78 (glucose-regulated protein 78) are correlated with the stability of atherosclerotic plaques.2 OBJECT: This research assesses changes in GRP78 and CHOP expressions in human umbilical vein endothelial cells(HUVEC) following experiments within the hemodynamic influencing factors test system. Additionally, it includes conducting an empirical study on the impact of blood flow shear force on the stability of human carotid atherosclerotic plaques. The objective is to explore the implications of blood flow shear force on the stability of carotid atherosclerotic plaques.
METHODS: The hemodynamic influencing factors test bench system was configured with low (Group A, 4 dyns/cm²), medium (Group B, 8 dyns/cm²), and high shear force groups (Group C, 12 dyns/cm²). Relative expression levels of GRP78 and CHOP proteins in human umbilical vein endothelial cells were measured using Western blot analysis, and quantitative analysis of GRP78 and CHOP mRNA was conducted using RT-qPCR. Meanwhile, plaques from 60 carotid artery patients, retrieved via Carotid Endarterectomy (CEA), were classified into stable (S) and unstable (U) groups based on pathological criteria. Shear force at the carotid bifurcation was measured preoperatively using ultrasound. Western blot and RT-qPCR were used to analyze the relative expression levels of GRP78 and CHOP proteins and mRNA, respectively, in the plaque specimens from both groups.
RESULTS: Expression levels of GRP78, CHOP proteins, and their mRNAs were assessed in groups A, B, and C via Western blot and RT-qPCR. Results showed that in the low-shear group, all markers were elevated in group A compared to groups B and C. Statistical analysis revealed significantly lower shear forces at the carotid bifurcation in group U compared to group S. In group U plaques, GRP78 and CHOP expressions were significantly higher in group U than in group S.
CONCLUSIONS: Blood flow shear forces variably affect the expression of GRP78 and CHOP proteins, as well as their mRNA levels, in vascular endothelial cells. The lower the shear force and fluid flow rate, the higher the expression of GRP78 and CHOP, potentially leading to endoplasmic reticulum stress(ERS), which may destabilize the plaque.
METHODS: The hemodynamic influencing factors test bench system was configured with low (Group A, 4 dyns/cm²), medium (Group B, 8 dyns/cm²), and high shear force groups (Group C, 12 dyns/cm²). Relative expression levels of GRP78 and CHOP proteins in human umbilical vein endothelial cells were measured using Western blot analysis, and quantitative analysis of GRP78 and CHOP mRNA was conducted using RT-qPCR. Meanwhile, plaques from 60 carotid artery patients, retrieved via Carotid Endarterectomy (CEA), were classified into stable (S) and unstable (U) groups based on pathological criteria. Shear force at the carotid bifurcation was measured preoperatively using ultrasound. Western blot and RT-qPCR were used to analyze the relative expression levels of GRP78 and CHOP proteins and mRNA, respectively, in the plaque specimens from both groups.
RESULTS: Expression levels of GRP78, CHOP proteins, and their mRNAs were assessed in groups A, B, and C via Western blot and RT-qPCR. Results showed that in the low-shear group, all markers were elevated in group A compared to groups B and C. Statistical analysis revealed significantly lower shear forces at the carotid bifurcation in group U compared to group S. In group U plaques, GRP78 and CHOP expressions were significantly higher in group U than in group S.
CONCLUSIONS: Blood flow shear forces variably affect the expression of GRP78 and CHOP proteins, as well as their mRNA levels, in vascular endothelial cells. The lower the shear force and fluid flow rate, the higher the expression of GRP78 and CHOP, potentially leading to endoplasmic reticulum stress(ERS), which may destabilize the plaque.
摘要：
背景：各种因素，包括血,炎症，传染性,和免疫因素，会导致缺血性中风。然而,主要原因通常是颈动脉硬化斑块的不稳定。据估计，18-25％的缺血性中风是由颈动脉斑块破裂引起的。1斑块稳定性对确定患者预后至关重要。开发一个高度准确的，非侵入性，或评估颈动脉斑块稳定性的微创技术对于诊断和治疗中风至关重要。我们小组先前的研究表明，CHOP（C/EBP同源蛋白）和GRP78（葡萄糖调节蛋白78）的表达水平与动脉粥样硬化斑块的稳定性相关。2目的：本研究评估了在血流动力学影响因素测试系统中进行实验后，GRP78和CHOP表达的变化。此外,它包括对血流剪切力对人颈动脉粥样硬化斑块稳定性的影响进行实证研究。目的是探讨血流剪切力对颈动脉粥样硬化斑块稳定性的影响。
方法：血液动力学影响因素测试台系统配置为低（A组，4达因/平方厘米)，中等(B组，8达因/平方厘米)，和高剪切力组(C组，12达因/平方厘米）。使用Westernblot分析测量人脐静脉内皮细胞中GRP78和CHOP蛋白的相对表达水平，使用RT-qPCR对GRP78和CHOPmRNA进行定量分析。同时,来自60例颈动脉患者的斑块，通过颈动脉内膜切除术（CEA）检索，根据病理标准分为稳定（S）和不稳定（U）组。术前使用超声测量颈动脉分叉处的剪切力。采用Westernblot和RT-qPCR分析GRP78和CHOP蛋白及mRNA的相对表达水平,分别,在两组的斑块标本中。
结果：GRP78，CHOP蛋白的表达水平，他们的mRNA在A组中进行了评估，B,和C通过Western印迹和RT-qPCR。结果表明，在低剪切组中，与B组和C组相比，A组的所有标志物均升高。统计学分析表明，与S组相比，U组的颈动脉分叉处的剪切力显着降低。在U组斑块中，U组GRP78和CHOP的表达明显高于S组。
结论：血流剪切力对GRP78和CHOP蛋白的表达有不同程度的影响，以及它们的mRNA水平，在血管内皮细胞中。剪切力和流体流速越低，GRP78和CHOP的表达越高，可能导致内质网应激（ERS），这可能会破坏斑块的稳定性。
方法：血液动力学影响因素测试台系统配置为低（A组，4达因/平方厘米)，中等(B组，8达因/平方厘米)，和高剪切力组(C组，12达因/平方厘米）。使用Westernblot分析测量人脐静脉内皮细胞中GRP78和CHOP蛋白的相对表达水平，使用RT-qPCR对GRP78和CHOPmRNA进行定量分析。同时,来自60例颈动脉患者的斑块，通过颈动脉内膜切除术（CEA）检索，根据病理标准分为稳定（S）和不稳定（U）组。术前使用超声测量颈动脉分叉处的剪切力。采用Westernblot和RT-qPCR分析GRP78和CHOP蛋白及mRNA的相对表达水平,分别,在两组的斑块标本中。
结果：GRP78，CHOP蛋白的表达水平，他们的mRNA在A组中进行了评估，B,和C通过Western印迹和RT-qPCR。结果表明，在低剪切组中，与B组和C组相比，A组的所有标志物均升高。统计学分析表明，与S组相比，U组的颈动脉分叉处的剪切力显着降低。在U组斑块中，U组GRP78和CHOP的表达明显高于S组。
结论：血流剪切力对GRP78和CHOP蛋白的表达有不同程度的影响，以及它们的mRNA水平，在血管内皮细胞中。剪切力和流体流速越低，GRP78和CHOP的表达越高，可能导致内质网应激（ERS），这可能会破坏斑块的稳定性。
