| 严蕊,邵轶群,王铁烽.王不留行黄酮苷通过NLRP3/Caspase-1 信号通路抑制ox-LDL诱导的细胞焦亡[J].浙江中医药大学学报,2023,47(11):1263-1269. |
| 王不留行黄酮苷通过NLRP3/Caspase-1 信号通路抑制ox-LDL诱导的细胞焦亡 |
| Vaccarin Attenuated Ox-LDL-induced HUVECs Pyroptosis by Suppressing NLRP3/Caspase-1 Signaling Pathway |
| DOI：10.16466/j.issn1005-5509.2023.11.004 |
| 中文关键词: 王不留行黄酮苷 ox-LDL HUVECs 焦亡 凋亡 炎症 |
| 英文关键词: Vaccarin ox-LDL HUVECs pyroptosis apoptosis inflammation |
| 基金项目:国家自然科学基金青年项目(82104720)；浙江省中医药科学研究基金项目(2020ZB266) |
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| 中文摘要: |
| [目的] 探讨王不留行黄酮苷对氧化低密度脂蛋白(oxidized low-density lipoprotein，ox-LDL)诱导的人脐静脉血管内皮细胞(human umbilical vein endothelial cells，HUVECs)损伤的保护作用及作用机制。[方法] 以ox-LDL(100 μg·mL-1)刺激HUVECs建立模型。采用细胞增殖活性检测(cell counting kit-8，CCK-8)法检测1～10 μmol·L-1王不留行黄酮苷对ox-LDL诱导的HUVECs的保护作用；采用乳酸脱氢酶(lactate dehydrogenase，LDH)法检测LDH的释放；采用Annexin Ⅴ-FITC/碘化丙啶(propidium iodide，PI)凋亡试剂盒检测细胞凋亡；使用2’，7’-二氯荧光素二乙酸酯(2’，7’-dichlorodihydrofluorescein diacetate，DCFD-A)荧光探针检测细胞内活性氧(reactive oxygen species，ROS)水平；采用实时荧光定量聚合酶链式反应(Real-time polymerase chain reaction，Real-time PCR)检测白细胞介素-6(interleukin-6，IL-6)、单核细胞趋化蛋白-1(monocyte chemoattracctant protein-1，MCP-1)、人血管内皮细胞黏附分子-1(human vascular cell adhesion molecule-1，VCAM-1)的mRNA表达；采用免疫印迹法检测B细胞淋巴瘤-2(B-cell lymphoma-2，Bcl-2)、Bcl-2相关x蛋白(Bcl-2 associated X protein，Bax)、NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3，NLRP3)、胱天蛋白酶-1(cysteinyl aspartate specific proteinase-1，caspase-1)、裂解的胱天蛋白酶-1(cleaved caspase-1)、IL-1β和IL-18的蛋白表达。[结果] 1～10 μg·mL -1王不留行黄酮苷减轻100 μg·mL-1的ox-LDL诱导的HUVECs损伤(P<0.05，P<0.01)。王不留行黄酮苷减轻ox-LDL诱导的LDH释放，减少细胞凋亡，同时促进Bcl-2蛋白表达，抑制Bax蛋白表达(P<0.01)。王不留行黄酮苷显著减少了ox-LDL诱导的ROS生成，抑制ox-LDL诱导的炎症因子IL-6、MCP-1和VCAM-1的水平(P<0.01)。王不留行黄酮苷抑制了ox-LDL诱导的NLRP3炎症小体介导的焦亡相关蛋白表达(P<0.05，P<0.01)。[结论] 王不留行黄酮苷能够减轻ox-LDL诱导的HUVECs凋亡，缓解炎症反应，其机制可能与NLRP3介导的焦亡有关。 |
| 英文摘要: |
| [Objective] To explore the protective effect and mechanism of Vaccarin on oxidized low-density lipoprotein(ox-LDL)-induced damage of human umbilical vein endothelial cells(HUVECs). [Methods] The protective effect of 1～10 μmol·L-1 Vaccarin on ox-LDL-induced HUVECs was detected by cell counting kit-8(CCK-8); the release of lactate dehydrogenase(LDH) was detected by LDH method; the cells apoptosis was detected using Annexin Ⅴ-FITC/propidium iodide(PI) apoptosis kit; 2'，7'-dichlorofluorescein diacetate(DCFD-A) fluorescent probe was used to detect intracellular reactive oxygen species(ROS) levels; Real-time quantitative polymerase chain reaction(Real-time PCR) was used to detect the mRNA expression of interleukin-6(IL-6)， monocyte chemoattractant protein-1(MCP-1) and vascular cell adhesion molecule-1(VCAM-1); Western blot was used to detect the protein expression of B-cell lymphoma-2(Bcl-2)， Bcl-2 associated X protein(Bax)， NOD-like receptor thermal protein domain associated protein 3(NLRP3)， cysteinyl aspartate specific proteinase-1(caspase-1)， cleaved caspase-1， IL-1β and IL-18. [Results] Vaccarin 1～10 μg·mL-1 ameliorated the damage of HUVECs induced by ox-LDL at 100 μg·mL -1(P<0.05， P<0.01). Vaccarin attenuated ox-LDL-induced LDH release and reduced apoptosis， while promoting Bcl-2 protein expression and inhibiting Bax protein expression(P<0.01). Vaccarin significantly reduced ox-LDL-induced ROS generation and inhibited ox-LDL-induced inflammatory factors IL-6， MCP-1 and VCAM-1 levels(P<0.01). Vaccarin inhibited ox-LDL-induced expression of NLRP3 inflammasome-mediated pyroptosis-related proteins(P<0.05， P<0.01). [Conclusion] Vaccarin could alleviate the apoptosis of HUVECs induced by ox-LDL and reduce the inflammatory response， and its mechanism may be tightly related to NLRP3 inflammasome activation-mediated pyroptosis. |
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