川芎嗪对补体旁路激活致内皮细胞炎症反应的干预作用
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摘要
目的研究川芎嗪(TMP)对补体旁路激活致人微血管内皮细胞(HMEC)炎症反应的干预作用及其可能的分子机制。方法以眼镜蛇毒因子(cobra venom factor,CVF)来激活正常人血清的补体旁路,用激活产物作用于HMEC,作用前加入不同浓度TMP预处理细胞,然后将细胞暴露在该激活产物中,采用ELISA方法检测细胞培养上清中可溶性黏附分子(ICAM-1、VCAM-1、E-selectin)和炎症介质(IL-6、TNF-α)的含量变化;双萤光素酶报告基因检测试剂盒测定NF-κB核内转录活性。结果 HMEC受补体旁路激活产物刺激后,引起黏附分子和炎症介质的表达上调,以及NF-κB核内转录活性的上调。不同浓度的TMP对上述炎症反应相关指标的上调均有干预作用,且表现出剂量依赖性。结论 TMP对补体旁路特异激活HMEC炎症反应有明显的干预作用,其机制可能与抑制NF-κB的核内转录活性有关。
Aim To study the intervention effect of tetramethylpyrazine(TMP) on human microvascular endothelial cells(HMECs) inflammatory response induced by activated complement alternative pathway and the possible molecular mechanisms.Methods HMECs were pretreated with different concentrations of TMP, and then exposed to the activated products of the complement alternative pathway which was prepared by cobra venom factor(CVF). The supernatant was removed and assayed for expression of the adhesion molecules(ICAM-1, VCAM-1 and E-selectin) and the inflammatory mediator(IL-6 and TNF-α) by using ELISA reagent kits. The nucleus transcriptional activity of NF-κB was measured by the dual luciferase reporter assay system.Results The adhesion molecules, inflammatory mediator and nucleus transcriptional activity of NF-κB increased after HMECs were exposed to the products of the activated complement alternative pathway.The up-regulation of ICAM-1, VCAM-1, E-selectin, IL-6, TNF-α and the nucleus transcriptional activity of NF-κB were inhibited by various concentrations of TMP in a dose-dependent manner.Conclusions TMP can effectively reduce inflammatory response of HMECs induced by the activated complement alternative pathway, and the mechanism may be highly related to inhibition of nucleus transcriptional activity of NF-κB.
Aim To study the intervention effect of tetramethylpyrazine(TMP) on human microvascular endothelial cells(HMECs) inflammatory response induced by activated complement alternative pathway and the possible molecular mechanisms.Methods HMECs were pretreated with different concentrations of TMP, and then exposed to the activated products of the complement alternative pathway which was prepared by cobra venom factor(CVF). The supernatant was removed and assayed for expression of the adhesion molecules(ICAM-1, VCAM-1 and E-selectin) and the inflammatory mediator(IL-6 and TNF-α) by using ELISA reagent kits. The nucleus transcriptional activity of NF-κB was measured by the dual luciferase reporter assay system.Results The adhesion molecules, inflammatory mediator and nucleus transcriptional activity of NF-κB increased after HMECs were exposed to the products of the activated complement alternative pathway.The up-regulation of ICAM-1, VCAM-1, E-selectin, IL-6, TNF-α and the nucleus transcriptional activity of NF-κB were inhibited by various concentrations of TMP in a dose-dependent manner.Conclusions TMP can effectively reduce inflammatory response of HMECs induced by the activated complement alternative pathway, and the mechanism may be highly related to inhibition of nucleus transcriptional activity of NF-κB.
引文
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[11] 李朝胜, 孙黔云. 三种化学小分子对补体旁路激活致内皮细胞黏附分子表达的干预及机制研究[J]. 中国药理学通报, 2015, 31(10): 1421-6.
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[12] 路青瑜, 李敏, 孙黔云. 补体旁路激活致内皮细胞纤溶凝血相关分子表达变化及干预研究[J]. 中国药理学通报, 2015, 31(8): 1142-6.
[12] Lu Q Y, Li M. Sun Q Y. Expression of coagulation-and fibrinolysis-related molecules of endothelial cells induced by activated complement alternative pathway and intervention[J]. Chin Pharmacol Bull, 2015, 31(8):1142-6.
[13] Morgan B P. The membrane attack complex as an inflammatory trigger[J]. Immunobiology, 2016, 221(6): 747-51.
[14] 鹿中华, 王锦权, 陶晓根, 等. 川芎嗪对脓毒症大鼠肝细胞线粒体保护作用的实验研究[J]. 中国中西医结合急救杂志, 2008, 15(2): 85-8.
[14] Lu Z H, Wang J Q, Tao X G, et al. An experimental study on protective effect of tetrmethylpyrazine on liver mitochondria in rat with sepsis[J].Chin J TCM WM Crit Care, 2008, 15(2): 85-8.
[15] 王国峰, 陆峰, 赵霞, 等. 川芎嗪对氧化低密度脂蛋白诱导内皮细胞炎症反应的影响[J].中华高血压杂志, 2012, 20(4): 347-51.
[15] Wang G F, Lu F, Zhao X, et al. Effects of ligustrazine on inflammation induced by oxidized low density lipoprotein in endothelial cells[J]. China J Hypertens, 2012, 20(4): 347-51.
[16] Tian J Q, Zhang W. Clinical study of adjuvant therapy on ischemic stroke with salviae miltiorrhizae and ligustrazine[J]. J Acute Dis, 2017, 6(1): 36-40.
[2] Bach L A. Endothelial cells and the IGF system[J]. J Mol Endocrinol, 2015, 54(1): R1-13.
[3] Macor P, Tedesco F. Complement as effector system in cancer immunotherapy[J]. Immunol Lett, 2007, 111(1): 6-13.
[4] 孙黔云. 眼镜蛇毒因子在生命科学中的应用[J]. 生命科学, 2016, 28(1): 22-6.
[4] Sun Q Y. Application of cobra venom factor in life sciences[J]. Chin Bull Life Sci, 2016, 28(1): 22-6.
[5] Dalvi P, Sun B, Tang N, Pulliam L. Immune activated monocyte exosomes alter microRNAs in brain endothelial cells and initiate an inflammatory response through the TLR4/MyD88 pathway[J]. Sci Rep-UK, 2017, 7(1): 1-12.
[6] 赵琳, 魏敏杰, 何苗,等.川芎嗪对阿尔采默病模型小鼠学习记忆能力的影响及其机制初探[J]. 中国药理学通报, 2008, 24(8): 1088-92.
[6] Zhao L, Wei M J, He M, et al. The effects of tetramethylpyrazine on learning and memory abilities of mice with Alzheimer disease and its possible mechanism[J]. Chin Pharmacol Bull, 2008, 24(8): 1088-92.
[7] 杨锡兰, 李言, 王盼, 等. 川芎嗪对脂多糖诱导的内皮细胞损伤的保护作用[J]. 蚌埠医学院学报, 2016, 41(3): 284-7.
[7] Yang X L, Li Y, Wang P, et al. Protective effect of tetramethylpyrazine on the injury endothelial cell induced by lipopolysaccharide[J]. J Bengbu Medi Coll, 2016, 41(3): 284-7.
[8] Feng L, Ke N, Cheng F, et al. The protective mechanism of ligustrazine against renal ischemia/reperfusion injury[J]. J Surg Res, 2011, 166(2): 298-305.
[9] 孙黔云, 王婉瑜, 熊郁良. 眼镜蛇毒高活性抗补体因子的体外溶血活性研究[J]. 中国药理学通报, 2003, 19(8):909-13.
[9] Sun Q Y, Wang W Y, Xiong Y L. In vitro hemolyzation of a highly active anticomplement factor from the venom of Naja kaouthia[J].Chin Pharmacol Bull, 2003, 19(8): 909-13.
[10] 孙黔云, 李敏, 叶巧玲, 李红玲. 补体旁路激活导致内皮细胞活化和损伤[J]. 中国药理学通报, 2012, 28(7): 925-9.
[10] Sun Q Y, Li M, Ye Q L, Li H L. Endothelial cell activation and injury induced by complement alternative pathway[J]. Chin Pharmacol Bull, 2012, 28(7): 925-9.
[11] 李朝胜, 孙黔云. 三种化学小分子对补体旁路激活致内皮细胞黏附分子表达的干预及机制研究[J]. 中国药理学通报, 2015, 31(10): 1421-6.
[11] Li C S, Sun Q Y. Effect of three chemical molecules on adhesion molecules expression in HMECs induced by activated complement alternative pathway[J]. Chin Pharmacol Bull, 2015, 31(10): 1421-6.
[12] 路青瑜, 李敏, 孙黔云. 补体旁路激活致内皮细胞纤溶凝血相关分子表达变化及干预研究[J]. 中国药理学通报, 2015, 31(8): 1142-6.
[12] Lu Q Y, Li M. Sun Q Y. Expression of coagulation-and fibrinolysis-related molecules of endothelial cells induced by activated complement alternative pathway and intervention[J]. Chin Pharmacol Bull, 2015, 31(8):1142-6.
[13] Morgan B P. The membrane attack complex as an inflammatory trigger[J]. Immunobiology, 2016, 221(6): 747-51.
[14] 鹿中华, 王锦权, 陶晓根, 等. 川芎嗪对脓毒症大鼠肝细胞线粒体保护作用的实验研究[J]. 中国中西医结合急救杂志, 2008, 15(2): 85-8.
[14] Lu Z H, Wang J Q, Tao X G, et al. An experimental study on protective effect of tetrmethylpyrazine on liver mitochondria in rat with sepsis[J].Chin J TCM WM Crit Care, 2008, 15(2): 85-8.
[15] 王国峰, 陆峰, 赵霞, 等. 川芎嗪对氧化低密度脂蛋白诱导内皮细胞炎症反应的影响[J].中华高血压杂志, 2012, 20(4): 347-51.
[15] Wang G F, Lu F, Zhao X, et al. Effects of ligustrazine on inflammation induced by oxidized low density lipoprotein in endothelial cells[J]. China J Hypertens, 2012, 20(4): 347-51.
[16] Tian J Q, Zhang W. Clinical study of adjuvant therapy on ischemic stroke with salviae miltiorrhizae and ligustrazine[J]. J Acute Dis, 2017, 6(1): 36-40.