Effects of Apelin-13/putative receptor protein related to AT1 homodimer on behaviors of human umbilical vein endothelial cells北大核心CSCD

Aim To explore the effects of putative receptor protein related to ATI (APJ) homodimer on the behaviors-the proliferation, migration and tube formation of human umbilical vein endothelial cells (HU-VECs). Methods HUVECs at logarithmic growth stage were randomly divided into PBS, Apelin-13 + TM1 (APJ monomer group) and Apelin-13 + PBS group (APJ homodimer group). Western blot and Matrix-Assisted Laser Desorption/Ionization Time of Fligh Mass Spectrometry (MALDI-TOF MS) were used to detect the expression of APJ and APJ homodimer in HUVECs, respectively. Real-Time Cell Analyzers (RT-CA) was used to detect the concentration of the maximum effect of Apelin-13. Cell viability was detected by CCK-8. The cell migration ability was detected by scratch test, and the number of tubes formed on matri-gel that made artificial basement membrane was counted. Results Western blot and MALDI-TOF MS showed that APJ and APJ homodimer were expressed in HUVECs. The EC50 of Apelin-13 was 2.26 x 10 -8 mol • L, and the concentration of the maximum effect was 1.0 X 10 mol • L. The results of CCK-8 experiment, migration experiment and tube formation showed that cells in each group migrated to the scratch bare area gradually with the extension of time, and the tube formation of cells in each group was also observed under the microscope. Data statistics showed that the proliferation, migration and tube forming abilities of HUVECs in Apelin-13 + PBS and Apelin-13 + TM1 groups were significantly higher than those in PBS group, and the abilities of proliferation, migration and tube forming in Apelin-13 + PBS group were significantly better than those in Apelin-13 + TM1 group (P < 0.05). Conclusions APJ homodimer can promote the proliferation, migration and tube formation of HUVECs, and the effects are better than APJ monomer. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Correlation between serum Dickkopf-1 (DKK1) levels and coronary artery stenosis

Background and aimsTo study the correlation between the level of serum Dickkopf-1 (DKK1) and the degree of coronary artery stenosis in patients with coronary atherosclerotic heart disease.Methods and resultsIn 2018, general data and biochemical indexes of 311 patients who underwent coronary angiography were recorded. Before procedure, arterial blood was drawn and the concentrations of DKK1, retinol binding protein 4 (RBP4), plasminogen activator inhibitor (PAI-1) were measured. Based on coronary angiography results, subjects were divided into a coronary heart disease (CHD) group; and a non-coronary heart disease (non-CHD)group. The CHD group was divided into three subgroups: the low Gensini score; the middle Gensini score; and the high Gensini score subgroups. Compared with those of the non-CHD group, DKK1, RBP4 and PAI-1 of the CHD group were significantly higher, while the OC was lower.DKK1,RBP4 and PAI-1 levels of the middle and high Gensini subgroups were significantly higher, compared with that of the low Gensini subgroup. Differences between osteocalcin (OC), beta-isomerized C-terminal telopeptidase (β-CTX), and 25(OH)₂D₃ of the three subgroups were not significant.Correlation between DKK1 and the inflammatory factors, RBP4 and PAI-1, was positive. Correlation between DKK1 and β - CTX, 25(OH)₂D₃ and OC was not significant. DKK1 was a risk factor for CHD. The degree of coronary artery stenosis was related to DKK1 concentration.ConclusionsSerum DKK1 levels in coronary heart disease patients were significantly higher, and positively correlated with the degree of coronary artery stenosis. DKK1 level is an independent risk factor for coronary heart disease.     

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

[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。