Long non-coding RNA THRIL promotes LPS-induced inflammatory injury by down-regulating microRNA-125b in ATDC5 cells

BackgroundOsteoarthritis is an age-related disorder of bone-joint that causes pain and disability in middle and older people. This study aimed to investigate the potential effects of long non-coding RNA (lncRNA) THRIL on lipopolysaccharide (LPS)-induced osteoarthritis cell injury model (ATDC5 cell inflammatory injury), as well as the possible internal molecular mechanisms.MethodsCell viability and apoptosis were assessed using CCK-8 assay and Guava Nexin assay, respectively. Cell transfection was conducted to change the expression of THRIL and microRNA-125b (miR-125b) in ATDC5 cells. qRT-PCR was performed to detect the expression of THRIL, miR-125b and pro-inflammatory cytokines IL-6, TNF-α and monocyte chemotactic protein 1 (MCP-1) in ATDC5 cells. ELISA was used to measure the concentrations of IL-6, TNF-α and MCP-1 in culture supernatant of ATDC5 cells. Finally, the protein expression of key factors involved in cell apoptosis, inflammatory response, JAK1/STAT3 and NF-κB pathways were evaluated using western blotting.ResultsLPS significantly induced ATDC5 cell inflammatory injury and up-regulated the expression of THRIL. Overexpression of THRIL aggravated the LPS-induced ATDC5 cell inflammatory injury. Suppression of THRIL had opposite effects. Moreover, THRIL negatively regulated the expression of miR-125b in ATDC5 cells. miR-125b participated in the effects of THRIL overexpression on LPS-induced ATDC5 cell inflammatory injury. Furthermore, overexpression of THRIL enhanced the LPS-induced JAK1/STAT3 and NF-κB pathways activation by down-regulating miR-125b.ConclusionTHRIL exerted pro-inflammatory roles in LPS-induced osteoarthritis cell injury model. Overexpression of THRIL promoted LPS-induced ATDC5 cell inflammatory injury by down-regulating miR-125b and then activating JAK1/STAT3 and NF-κB pathways.     

脑心通胶囊药效成分对人脐静脉内皮细胞JAK/STAT信号通路、血管活性物质、黏附分子、炎症因子的影响

目的:研究脑心通胶囊主要药效成分对人脐静脉内皮细胞(HUVEC)JAK/STAT信号通路、血管活性物质、黏附分子、炎症因子的影响,以期阐明脑心通胶囊活血化瘀的作用机制.方法:采用CCK-8法检测不同浓度的12个脑心通胶囊药效成分[咖啡酸(1.56～200μmol/L)、阿魏酸(1.56～200μg/mL)、洋川芎内酯H...     

灯盏花乙素对非小细胞肺癌细胞JAK/STAT信号通路的影响

目的 研究灯盏花乙素对非小细胞肺癌细胞JAK/STAT信号通路及其凋亡机制的影响。方法 培养非小细胞肺癌A549细胞,经不同浓度灯盏花乙素处理24 h后收集细胞。采用MTT法观察细胞增殖,流式细胞术检测细胞凋亡;RT-qPCR法检测JAK2,STAT3,Pim1和Bcl-2的mRNA表达情况;Western blot检测JAK2和STAT3蛋白表达水平。结果 MTT法和流式细胞术结果显示,灯盏花乙素能降低A549细胞的存活率,促进其凋亡,并呈剂量依赖效应;RT-qPCR和Western blot结果显示,与对照组比较,灯盏花乙素各组JAK2,STAT3,Pim1和Bcl-2的mRNA表达降低,JAK2和STAT3蛋白水平下降,且与加入的灯盏花乙素浓度成反比。结论 灯盏花乙素可以促进非小细胞肺癌A549细胞凋亡,其机制可能是通过抑制JAK/STAT信号转导通路的激活,减少JAK2和STAT3蛋白的表达。     

微RNA-182-5p激动剂和抑制剂对肝癌细胞增殖和迁移的影响

目的研究微RNA(miR)-182-5p激动剂和抑制剂对肝癌细胞增殖和迁移的影响。方法采用实时荧光定量-聚合酶链反应(RT-qPCR)和蛋白质印迹法检测非肝细胞癌(HCC)肝组织,HCC组织及邻近组织标本中miR-182-5p的表达水平以及RND3的mRNA/蛋白表达水平。建立患者来源的HCC细胞培养物,并进行miR-182-5p激动剂和抑制剂转染处理,以分别模拟miRNA的过表达和敲减。通过Transwell细胞侵袭实验监测体外HCC细胞的迁移。通过细胞活力和增殖评价体外HCC细胞的生长。结果与非HCC或邻近的HCC组织相比,HCC组织中的miR-182-5p明显上调,与RND3 mRNA表达呈负相关。使用miR-182-5p激动剂可显著降低HCC细胞中RND3 mRNA/蛋白质表达水平。通过荧光素酶试验和顺乌头酸酶2(AGO2)-RNA免疫沉淀分析,验证了miR-182-5p对RND3 mRNA具有靶向作用。MiR-182-5p激动剂可显著降低RND3表达,促进HCC细胞体外迁移和侵袭,可能是通过Rho相关卷曲螺旋蛋白激酶1(ROCK1)和ROCK2的抑制来实现。结论miR-182-5p可以通过靶向RND3来提高体外HCC细胞的迁移和增殖。使用miR-182-5p抑制剂,可以抑制体外肝癌细胞的迁移和增殖。     

Astragalus polysaccharides suppress ICAM-1 and VCAM-1 expression in TNF-α-treated human vascular endothelial cells by blocking NF-κB activation

Aim:

To investigate the effects Astragalus polysaccharides (APS) on tumor necrosis factor (TNF)-α-induced inflammatory reactions in human umbilical vein endothelial cells (HUVECs) and to elucidate the underlying mechanisms.

Methods:

HUVECs were treated with TNF-α for 24 h. The amounts of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were determined with Western blotting. HUVEC viability and apoptosis were detected using cell viability assay and Hoechst staining, respectively. Reactive oxygen species (ROS) production was measured by DHE staining. Monocyte and HUVEC adhesion assay was used to detect endothelial cell adhesive function. NF-κB activation was detected with immunofluorescence.

Results:

TNF-α (1-80 ng/mL) caused dose- and time-dependent increases of ICAM-1 and VCAM-1 expression in HUVECs, accompanied by significant augmentation of IκB phosphorylation and NF-κB translocation into the nuclei. Pretreatment with APS (10 and 50 μg/mL) significantly attenuated TNFα-induced upregulation of ICAM-1 VCAM-1 and NF-κB translocation. Moreover, APS significantly reduced apoptosis, ROS generation and adhesion function damage in TNF-α-treated HUVECs.

Conclusion:

APS suppresses TNFα-induced adhesion molecule expression by blocking NF-κB signaling and inhibiting ROS generation in HUVECs. The results suggest that APS may be used to treat and prevent endothelial cell injury-related diseases.     

Schisandrin B inhibits LPS-induced inflammatory response in human umbilical vein endothelial cells by activating Nrf2

Schisandrin B (SchB), an active ingredient extracted from Schisandra chinensis (Turcz.) Baill, has been known to have anti-oxidant and anti-inflammatory activities. In this study, we investigated the anti-inflammatory effects and mechanism of SchB in LPS-stimulated human umbilical vein endothelial cells (HUVECs). The effects of SchB on VCAM-1, ICAM-1, NF-κB and Nrf2 expression were detected by western blot analysis. The effects of SchB on TNF-α and IL-8 production were detected by ELISA. The results showed that SchB strongly suppressed the production of TNF-α and IL-8 in HUVECs stimulated with LPS. SchB also inhibited LPS-induced VCAM-1 and ICAM-1 expression. Furthermore, SchB blocked the activation of NF-κB induced by LPS. In addition, SchB increased the expression of Nrf2 and HO-1 in a concentration-dependent manner. And the inhibition of TNF-α and IL-8 production by SchB was blocked by transfection with Nrf2 siRNA. Our findings showed that SchB inhibited LPS-induced inflammation in HUVECs by activating Nrf2 signaling pathway.     

Motorcycle exhaust particles up-regulate expression of vascular adhesion molecule-1 and intercellular adhesion molecule-1 in human umbilical vein endothelial cells

Epidemiological studies have shown that there is a strong correlation between atherosclerosis and ambient air pollution. In this study, we found that motorcycle exhaust particles (MEP) induced adhesion between cells of the human monocytic leukemia cell line (THP-1) and human umbilical vein endothelial cells (HUVECs) in a time-and dose-dependent manner. In addition, MEP treatment induced both mRNA and protein expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) in HUVECs. The IκB degradation and p65 nuclear translocation was found in MEP-treated HUVECs, suggested the involvement of nuclear factor-κB (NF-κB). MEP-induced VCAM-1 and ICAM-1 protein expression was inhibited by NF-κB inhibitor BAY 11-7085. Oxidative stress was also involved in the signaling of VCAM-1 and ICAM-1 expression. MEP treatment caused hydrogen peroxide and superoxide formation. Pretreatment with α-tocopherol could inhibit MEP-induced reactive oxygen intermediates generation and suppressed MEP-induced IκB degradation and adhesion molecules expression. Furthermore, the carbon black (CB) nanoparticles with different diameters could induce VCAM-1 and ICAM-1 protein expression; however, polycyclic aromatic hydrocarbons (PAHs) only increased the expression of ICAM-1 but not that of VCAM-1 in HUVECs. In this study, we found that MEPs could induce ICAM-1 and VCAM-1 expression through oxidative stress and NF-κB activation in HUVECs.     

二苯乙烯苷对H_2O_2诱导血管内皮细胞黏附分子表达的影响

目的研究二苯乙烯苷(TSG)对过氧化氢(H2O2)诱导损伤的人脐静脉内皮细胞P-selectin、E-selectin、ICAM-1、VCAM-1和MCP-1表达的影响。方法体外培养人脐静脉内皮细胞,实验分为空白对照组、H2O2组、辛伐他汀组、TSG组,运用逆转录聚合酶链式反应和酶联免疫吸附试验分别检测P-selectin、E-selectin、ICAM-1、VCAM-1和MCP-1 mRNA与蛋白的表达。结果 200μmol·L-1的H2O2作用内皮细胞24 h后,P-selectin、E-selectin、ICAM-1、VCAM-1和MCP-1的mRNA和蛋白表达水平均明显上调;经TSG预处理内皮细胞4 h后,再加200μmol·L-1的H2O2作用内皮细胞24 h,结果显示:TSG能抑制H2O2诱导的内皮细胞P-selnecti、E-se-lectin、ICAM-1、VCAM-1和MCP-1的mRNA和蛋白的表达。结论 TSG可抑制H2O2诱导的人脐静脉内皮细胞黏附分子的表达。     

阿托伐他汀对内皮细胞微粒诱导人脐静脉内皮细胞VCAM-1和ICAM-1表达的影响

目的:探讨阿托伐他汀对内皮细胞微粒(EMPs)诱导的人脐静脉内皮细胞(HUVECs)表达血管细胞粘附分子(VCAM)-1和细胞间粘附分子(ICAM)-1的影响。方法:取生长良好的第4,5代人脐静脉内皮细胞,将细胞分为3大组:对照组、EMPs组、EMPs+阿托伐他汀组。对照组加入培养基,EMPs组以不同浓度的EMPs(0/mL,1×102/mL,1×103/mL,1×104/mL,1×105/mL)与HUVECs共同孵育24 h,EMPs+阿托伐他汀组以不同浓度的阿托伐他汀(0.05,0.1,1.0,10μmol.L-1)与HUVECs作用1 h后,加入105/mL EMPs共同孵育24 h。分别采用实时荧光定量聚合酶链反应和蛋白免疫印迹方法检测VCAM-1和ICAM-1 mRNA和蛋白的表达。结果:HUVECs受EMPs刺激后,VCAM-1和ICAM-1 mRNA及蛋白表达呈浓度依赖性增加,阿托伐他汀可不同程度上抑制EMPs的作用。结论:阿托伐他汀抗动脉粥样硬化作用可能部分与抑制EMPs诱导的内皮细胞VCAM-1和ICAM-1的表达有关。     

s-油酰丙醇胺对人脐静脉内皮细胞黏附分子表达的影响

目的探讨s-油酰丙醇胺对用肿瘤坏死因子α(TNF-α)诱导的人脐静脉内皮细胞黏附分子(VCAM-1,I-CAM-1,E选择素)表达的影响。方法从新鲜的脐带中分离出人脐静脉内皮细胞,培养至3~9代,用不同浓度的s-油酰丙醇胺(10,50,100μmol/L)孵育12 h后,用TNF-α(20 ng/mL)孵育8 h,采用荧光实时定量PCR和细胞酶联免疫吸附试验分别检测VCAM-1,ICAM-1,E选择素的mRNA及蛋白的表达,同时采用细胞黏附实验检测其对细胞黏附的影响。结果相对于正常的人脐静脉内皮细胞,TNF-α诱导后的人脐静脉内皮细胞黏附分子(VCAM-1,ICAM-1,E-选择素)的表达明显增加。s-油酰丙醇胺可以显著的抑制VCAM-1的表达,并呈现出一定的剂量依赖性,而且对人急性单核细胞性白血病细胞(THP-1)的黏附也有明显的抑制作用,但对ICAM-1,E-选择素的表达却没有影响。结论s-油酰丙醇胺和大多数的PPARα激动剂一样,能够抑制慢性炎症,减少单核细胞的黏附,抑制VCAM-1的表达,而对急性炎症没有作用,如对E-选择素的表达无影响。     

Acanthoic acid inhibits LPS-induced inflammatory response by activating LXRα in human umbilical vein endothelial cells

Acanthoic acid, a pimaradiene diterpene isolated from Acanthopanax koreanum, has been reported to have anti-inflammatory activities. However, the effect of acanthoic acid on vascular inflammation has not been investigated. The aim of this study was to investigate the anti-inflammatory effects of acanthoic acid on lipopolysaccharide (LPS)-induced inflammatory response in human umbilical vein endothelial cells (HUVECs). The production of cytokines TNF-α and IL-8 was detected by ELISA. The expression of VCAM-1, ICAM-1, E-selectin, NF-κB and LXRα were detected by Western blotting. Adhesion of monocytes to HUVECs was detected by monocytic cell adhesion assay. The results showed that acanthoic acid dose-dependently inhibited LPS-induced TNF-α and IL-8 production. Acanthoic acid also inhibited TNF-α-induced IL-8 and IL-6 production. LPS-induced endothelial cell adhesion molecules, VCAM-1 and ICAM-1 were also inhibited by acanthoic acid. Acanthoic acid inhibited LPS-induced NF-κB activation. Furthermore, acanthoic acid dose-dependently up-regulated the expression of LXRα. In addition, our results showed that the anti-inflammatory effect of acanthoic acid was attenuated by transfection with LXRα siRNA. In conclusion, the anti-inflammatory effect of acanthoic acid is due to its ability to activate LXRα. Acanthoic acid may be a therapeutic agent for inflammatory cardiovascular disease.     

miR-149抑制结直肠癌细胞迁移的分子机制研究

目的 探讨miR-149对结直肠癌（CRC）细胞增殖、侵袭、迁移及凋亡的影响及其作用机制。方法 实时荧 光定量PCR（q-PCR）方法检测miR-149在CRC细胞SW620、LS174T和人结肠上皮细胞FHC中的表达水平;荧光素酶 分析法验证 STAT3 与 miR-149 之间的靶向结合关系;q-PCR 和 Western blot 检测 miR-149 过表达对 CRC 细胞中 STAT3、p-STAT3 mRNA和蛋白表达的影响。将miR-149 mimics与STAT3过表达质粒单独或者共转染至CRC细胞 中,并分为miR-NC组、miR-149 mimics组、miR-149 mimics+pEGFP/STAT3组。采用CCK-8法、Transwell法、细胞划 痕法、流式细胞术分别检测各分组CRC细胞的增殖、侵袭、迁移和凋亡情况。结果 与正常结肠上皮细胞FHC相比, CRC 细胞 SW620、LS174T 中 miR-149 表达水平受到抑制（P＜0.01）。荧光素酶实验证实,在 CRC 细胞中 STAT3 是 miR-149的一个直接靶基因。过量表达miR-149抑制STAT3、p-STAT3 mRNA和蛋白的表达,降低CRC细胞增殖能 力、侵袭能力以及迁移能力（P＜0.01）。同时,过量表达 miR-149 也可促进 CRC 细胞的凋亡（P＜0.01）。但过表达 STAT3可解除miR-149对CRC细胞增殖、侵袭以及迁移能力的抑制作用。结论 miR-149通过靶向STAT3抑制结直 肠癌细胞的增殖、侵袭与迁移,同时促进细胞的凋亡。     

Rhein attenuates renal inflammatory injury of uric acid nephropathy via lincRNA-Cox2/miR-150-5p/STAT1 axis

Rhein has protective effect on uric acid nephropathy (UAN). This article aims to demystify the mechanism of function of rhein in UAN. Mouse kidney epithelial cell line (TCMK-1) was incubated with uric acid (UA) to induce inflammatory injury. Then, the TCMK-1 cells were treated with rhein. The relationships among lincRNA-Cox2, miR-150-5p and STAT1 were evaluated by luciferase reporter assay. CCK8 and flow cytometry were performed to detect cell proliferation and apoptosis. The levels of IL-6, IL-1β and TNF-α were investigated by enzyme linked immunosorbent assay. Western blot and quantitative real-time PCR were performed to examine the expression of genes and proteins. We found that UA suppressed proliferation and enhanced apoptosis and the levels of IL-6, IL-1β and TNF-α of TCMK-1 cells, which was effectively improved by rhein treatment. Furthermore, lincRNA-Cox2 overexpression caused an increase of apoptosis and inflammatory factors in the rhein-treated TCMK-1 cells. LincRNA-Cox2 regulated STAT1 expression by sponging miR-150-5p. And lincRNA-Cox2 promoted apoptosis and inflammatory injury of TCMK-1 cells by regulating miR-150-5p/STAT1 axis. In summary, our studies demonstrate that rhein has a protective effect against UAN by inhibiting renal inflammatory injury via lincRNA-Cox2/miR-150-5p/STAT1 axis.     

Hesperidin, hesperidin methyl chalone and phellopterin from Poncirus trifoliata (Rutaceae) differentially regulate the expression of adhesion molecules in tumor necrosis factor-alpha-stimulated human umbilical vein endothelial cells

The fruits of Poncirus trifoliata (L.) are widely used in Oriental medicine to treat allergic inflammation. Recently, several active compounds including hesperidin, hesperidin methyl chalone and phellopterin from P. trifoliata (Rutaceae) were isolated and characterized. The goal of this study was to investigate the differential effect of hesperidin, hesperidin methyl chalone and phellopterin derived from P. trifoliata (Rutaceae) on the induction of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) by TNF-alpha and the possible molecular mechanisms by which they differentially regulate ICAM-1 and VCAM-1 expressions. Stimulation of human umbilical vein endothelial cells (HUVECs) with TNF-alpha resulted in the increase of ICAM-1 and VCAM-1 expressions, while pretreatment with the three components completely inhibited VCAM-1 expression in a dose-dependent manner but had no effect on ICAM-1 expression. All three compounds failed to block TNF-alpha-induced phosphorylation of ERK1/2, which is involved in regulating ICAM-1 production by TNF-alpha. Furthermore, they efficiently inhibited the phosphorylation of Akt and PKC, suggesting that Akt or PKC pathways are an important target by which these compounds regulate TNF-alpha-induced VCAM-1 but not ICAM-1. Additionally, treatment with these chemicals also inhibited U937 monocyte adhesion to HUVECs stimulated with TNF-alpha. Interestingly, the inhibitory effect of hesperidin, hesperidin methyl chalone and phellopterin on monocyte adhesion to HUVECs was recapitulated by transfecting cells with VCAM-1 siRNA. Taken together, hesperidin, hesperidin methyl chalone and phellopterin reduce TNF-alpha-induced VCAM-1 expression through regulation of the Akt and PKC pathway, which contributes to inhibit the adhesion of monocytes to endothelium.     

下调miR-106a-5p对过氧化氢诱导的心肌细胞损伤及JAK1/STAT3通路的影响

目的 探讨下调miR-106a-5p对过氧化氢(H₂O₂)诱导的心肌细胞损伤及Janus激酶1(JAK1)/信号转导和转录激活因子3(STAT3)通路的影响。方法 不同浓度梯度H₂O₂(0、50、100、200、400μmol/L)处理大鼠心肌细胞H9c2。H9c2细胞分为对照组、H₂O₂组(100μmol/L H₂O₂)、miR-106a-5p NC组(100μmol/L H₂O₂+转染miR-106a-5p NC)和miR-106a-5p siRNA组(100μmol/L H₂O₂+转染miR-106a-5p siRNA)。实时荧光定量PCR(qPCR)法检测miR-106a-5p表达;CCK-8法检测细胞增殖;流式细胞仪检测细胞凋亡;试剂盒检测细胞培养液中乳酸脱氢酶(LDH)含量、谷胱甘肽过氧化物酶(GSH-Px)活性及...     

长链非编码前列腺癌相关转录因子 6靶向微小 RNA-139-3p对骨肉瘤细胞增殖和凋亡的影响

目的 探讨长链非编码RNA(LncRNA)前列腺癌相关转录因子6(PCAT6)对骨肉瘤细胞增殖和凋亡的影响和分子机制.方法 实时荧光定量PCR(RT-qPCR)检测20例骨肉瘤组织和与其对应的癌旁组织中PCAT6和微小RNA-139-3p(miR-139-3p)的表达水平.双荧光素酶报告基因实验和RT-qPCR验证PCAT6对miR-139-3p的靶向调控关系.将PCAT6小干扰RNA(si-PCAT6)、miR-139-3p模拟物(miR-139-3p mimics)分别转染骨肉瘤细胞SAOS2,细胞计数试剂盒(CCK-8)检测SAOS2细胞增殖活力,流式细胞术检测SAOS2细胞凋亡,蛋白质印迹法检测B细胞淋巴瘤/白血病-2(Bcl-2)、细胞周期素D1(Cyclin D1)、Bcl-2相关X蛋白(Bax)和P21、的表达水平.将si-PCAT6和miR-139-3p抑制物(anti-miR-139-3p)共转染至SAOS2细胞,采用上述方法检测细胞增殖、迁移侵袭能力以及凋亡变化.结果 与瘤旁组织相比,骨肉瘤组织中PCAT6的表达水平[(2.76±0.27)比(1.01±0.09)]显著升高,miR-139-3p的表达水平[(0.51±0.05)比(1.00±0.08)]显著降低(P<0.05).miR-139-3p是PCAT6的靶基因,PCAT6靶向负性调控miR-139-3p表达.抑制PCAT6表达或过表达miR-139-3p均可下调SAOS2细胞CyclinD1和Bcl-2蛋白表达,上调p21和Bax蛋白表达,降低细胞活力,促进细胞凋亡(P<0.05).抑制miR-139-3p表达可逆转si-PCAT6对骨肉瘤SAOS2细胞增殖抑制和凋亡的影响(P<0.05).结论 lncRNA PCAT6在骨肉瘤组织中表达上调,抑制miR-139-3p通过靶向miR-139-3p抑制骨肉瘤细胞增殖,诱导骨肉瘤细胞凋亡.     

藁本内酯通过调控miR-133b-5p表达对脂多糖所致心肌损伤的保护作用

目的： 探讨藁本内酯（LIG）对脂多糖（LPS）所致心肌损伤的影响及分子机制。方法： 用质量浓度为10 mg·L^-1的LPS处理心肌细胞,记为LPS组,正常培养的细胞为对照（Con）组;用浓度分别为10,20,40 μmol·L^-1的藁本内酯处理心肌细胞后再用10 mg·L^-1的LPS处理,作为藁本内酯低、中、高浓度组;将miR-NC、miR-133b-5p转染至心肌细胞中再用10 mg·L^-1的LPS处理,记为LPS+miR-NC组、LPS+miR-133b-5p组;将anti-miR-NC、anti-miR-133b-5p转染至心肌细胞中再用40 μmol·L^-1的藁本内酯、10 mg·L^-1的LPS处理,记为LPS+LIG+anti-miR-NC组、LPS+LIG+anti-miR-133b-5p组。酶联免疫吸附法（ELISA）法检测肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）水平;流式细胞术检测细胞凋亡;蛋白质印迹（Western blot）法检测B细胞淋巴瘤/白血病-2（Bcl-2）、Bcl-2相关X蛋白（Bax）蛋白表达;实时荧光定量PCR （RT-qPCR）检测miR-133b-5p表达水平。结果： 藁本内酯低、中、高浓度处理后,LPS诱导的心肌细胞中TNF-α、IL-6水平显著降低,细胞凋亡率显著降低,Bcl-2表达水平显著升高,Bax表达水平显著降低,miR-133b-5p表达水平显著升高,且呈浓度依赖性（P<0.05）。miR-133b-5p过表达可抑制LPS诱导的心肌细胞中TNF-α、IL-6水平和细胞凋亡。抑制miR-133b-5p表达逆转了藁本内酯对LPS所致的心肌细胞损伤的保护作用。结论： 藁本内酯通过上调miR-133b-5p表达可抑制LPS诱导的心肌细胞凋亡和炎症反应,对LPS诱导的心肌损伤具有保护作用。     

Myristicin regulates proliferation and apoptosis in oxidized low-density lipoprotein-stimulated human vascular smooth muscle cells and human umbilical vein endothelial cells by regulating the PI3K/Akt/NF-κB signalling pathway

ContextAtherosclerosis (AS) is a chronic inflammatory disease. Human vascular smooth muscle cell (hVSMC) accumulation and human umbilical vein endothelial cell (HUVEC) dysfunction are associated with the pathogenesis of AS. This study explores whether myristicin plays a protective role in AS.Materials and methodshVSMCs and HUVECs were stimulated with 100 μg/mL oxidized low-density lipoprotein (ox-LDL) to establish a cellular model of AS. Cell viability, lactate dehydrogenase (LDH) release and cell apoptosis were evaluated using MTT, LDH and flow cytometry assays, respectively. Cell migration and inflammatory cytokine release were assessed using Transwell assay and ELISA.ResultsMyristicin (5, 10, 25, and 50 μM) had no obvious effect on cell viability or the activity of LDH in hVSMCs, while 100 and 200 μM myristicin markedly suppressed hVSMCs viability and increased LDH release. Myristicin had no obvious effect on cell viability or the activity of LDH in HUVECs. Myristicin inhibited viability and increased apoptosis in ox-LDL-treated hVSMCs, but was associated with increased proliferation and inhibited apoptosis in HUVECs stimulated by ox-LDL. Additionally, myristicin markedly suppressed ox-LDL-induced hVSMCs migration and the release of inflammatory cytokines, including MCP-1, IL-6, VCAM-1 and ICAM-1, in HUVECs. Results also demonstrated that the promoting effects of ox-LDL on the PI3K/Akt and NF-κB signalling pathway in both hVSMCs and HUVECs were abolished by treatment with myristicin.Discussion and conclusionsMyristicin regulated proliferation and apoptosis by regulating the PI3K/Akt/NF-κB signalling pathway in ox-LDL-stimulated hVSMCs and HUVECs. Thus, myristicin may be used as a new potential drug for AS treatment.