Lin Yen-Kuang, Hsieh Tsung-Han, Yeh Chi-Tai, Yadav Vijesh Kumar, Fong Iat-Hang, Kuo Kuang-Tai, Kounis Nicholas G, Hu Patrick, Hung Ming-Yow
Graduate Institute of Athletics and Coaching Science, National Taiwan Sport University, No. 250 Wenhua 1st Rd., Guishan, Taoyuan 33301, Taiwan.
Biostatistics Research Center, Taipei Medical University, Taipei City 11031, Taiwan.
Pharmaceuticals (Basel). 2025 Sep 16;18(9):1384. doi: 10.3390/ph18091384.
Lipoprotein(a) [Lp(a)]-induced inflammation contributes to coronary artery spasm (CAS) by the contraction of vascular smooth muscle cells. However, the interaction between Lp(a) and soluble CD36 (sCD36)/interleukin (IL)-6/RAS Homolog Family Member A (RhoA)-GTP signaling pathway has not been evaluated. We investigated the relevance of Lp(a)/CD36 signaling in CAS patient monocyte-derived macrophages (PMDMs) and a human coronary artery smooth muscle cell (HCASMC) line using expression profile correlation analyses, molecular docking, RNA sequencing, flow cytometry, immunoblotting, and quantitative reverse transcription polymerase chain reaction. Plasma Lp(a) and sCD36 levels in 41 CAS patients were significantly higher ( = 0.001) and positively correlated (r = 0.3145, < 0.001), a trend not observed in 36 non-CAS controls. RNA sequencing indicated a significant co-overexpression of CD36 and RhoA in Lp(a)-treated CAS PMDMs and HCASMCs, of which the mRNA and protein expression of CD36 and RhoA were significantly enhanced ( < 0.001) dose-dependently. Lp(a) rather than LDL preferentially induced CD80+ PMDM (M1) polarization. In HCASMCs, the CD36 knockdown using either short hairpin RNA or natural biflavonoid amentoflavone suppressed Lp(a)-upregulated protein expression of CD36, RhoA-GTP, IL-6, tumor necrosis factor (TNF)-α, nuclear factor (NF)-κB, and CD80; however, overexpressed CD36 increased their levels. Lp(a) decreased and amentoflavone increased the epigenetic expression of CD36 inhibitors, miR-335-5p, and miR-448, respectively. Reciprocally, an miRNA inhibitor or mimic could magnify or diminish Lp(a)-induced CD36, TNF-α, NF-κB and IL-6 expressions in HCASMCs, respectively. Elevated Lp(a) levels upregulate the CD36-dependent TNF-α/NF-κB/IL-6/RhoA-GTP signaling pathway in CAS PMDMs and HCASMCs, indicating that Lp(a)/CD36 inflammatory signaling, HCASMC activation, and macrophage M1 polarization mediate CAS development.
脂蛋白(a)[Lp(a)]诱导的炎症通过血管平滑肌细胞收缩导致冠状动脉痉挛(CAS)。然而，Lp(a)与可溶性CD36(sCD36)/白细胞介素(IL)-6/RAS同源家族成员A(RhoA)-GTP信号通路之间的相互作用尚未得到评估。我们使用表达谱相关分析、分子对接、RNA测序、流式细胞术、免疫印迹和定量逆转录聚合酶链反应，研究了Lp(a)/CD36信号在CAS患者单核细胞衍生巨噬细胞(PMDMs)和人冠状动脉平滑肌细胞(HCASMC)系中的相关性。41例CAS患者的血浆Lp(a)和sCD36水平显著更高(P = 0.001)且呈正相关(r = 0.3145，P < 0.001)，36例非CAS对照中未观察到这种趋势。RNA测序表明，在Lp(a)处理的CAS PMDMs和HCASMCs中，CD36和RhoA显著共过表达，其中CD36和RhoA的mRNA和蛋白表达呈剂量依赖性显著增强(P < 0.001)。Lp(a)而非低密度脂蛋白(LDL)优先诱导CD80+ PMDM(M1)极化。在HCASMCs中，使用短发夹RNA或天然双黄酮穗花杉双黄酮敲低CD36可抑制Lp(a)上调的CD36、RhoA-GTP、IL-6、肿瘤坏死因子(TNF)-α、核因子(NF)-κB和CD80的蛋白表达；然而，过表达CD36会增加它们的水平。Lp(a)分别降低和穗花杉双黄酮增加CD36抑制剂miR-335-5p和miR-448的表观遗传表达。相反，miRNA抑制剂或模拟物可分别放大或减弱Lp(a)诱导的HCASMCs中CD36、TNF-α、NF-κB和IL-6的表达。升高Lp(a)水平上调CAS PMDMs和HCASMCs中CD36依赖性TNF-α/NF-κB/IL-6/RhoA-GTP信号通路，表明Lp(a)/CD36炎症信号、HCASMC激活和巨噬细胞M1极化介导CAS发展。
