Guangdong Pharmaceutical University, Guangzhou Higher Education Mega Center, Guangzhou, Guangdong 510006, P.R. China.
Department of Cardiology, The First Affiliated Hospital of Sun Yat‑sen University, Guangzhou, Guangdong 510080, P.R. China.
Mol Med Rep. 2018 Apr;17(4):5109-5115. doi: 10.3892/mmr.2018.8488. Epub 2018 Jan 25.
Polysaccharide from Fuzi (FPS) is a water‑soluble polysaccharide isolated from the traditional Chinese herbal medicine Fuzi. It has been demonstrated to protect hepatocytes against ischemia‑reperfusion injury through its potent antioxidant effects, and to attenuate starvation‑induced cytotoxicity in H9c2 cells by increasing autophagic activity. In the present study, Alizarin Red S staining was used to detect mineral deposition and reverse transcription‑quantitative polymerase chain reaction was used to detect the core binding factor α1 and smooth muscle 22α mRNA expression. To analyze autophagic activity, western blotting was used to detect microtubule‑associated protein 1A/1B light chain 3 and nucleoporin P62 expression. In addition, green fluorescent protein‑LC3 dots‑per‑cell was observed by fluorescence microscopy. It was demonstrated that oxidized low‑density lipoprotein (Ox‑LDL) could increase the calcification of human vascular smooth muscle cells (VSMCs) in a concentration‑dependent manner, and that FPS treatment had a significant protective effect against Ox‑LDL‑induced calcification of human VSMCs. Furthermore, FPS treatment alleviated the Ox‑LDL‑induced downregulation of autophagic activity, and the protective effect of FPS on Ox‑LDL‑induced calcification was attenuated by the autophagy inhibitor 3‑methyladenine. In conclusion, the present study demonstrated for the first time to the best of the authors' knowledge that FPS can protect against Ox‑LDL‑induced vascular calcification in human VSMCs, and that this likely occurs via the activation of autophagy. This supports the hypothesis that autophagy may be an endogenous protective mechanism counteracting vascular calcification, and that FPS may be used as a potential therapeutic for vascular calcification.
从中药附子中分离得到的水溶性多糖附子多糖(FPS)已被证明具有很强的抗氧化作用,可保护肝细胞免受缺血再灌注损伤,并通过增加自噬活性来减轻 H9c2 细胞饥饿诱导的细胞毒性。在本研究中,使用茜素红 S 染色检测矿化沉积,逆转录定量聚合酶链反应检测核心结合因子α1和平滑肌 22α mRNA 表达。分析自噬活性时,使用 Western blot 检测微管相关蛋白 1A/1B 轻链 3 和核孔蛋白 P62 的表达。此外,通过荧光显微镜观察绿色荧光蛋白-LC3 斑点/细胞。结果表明,氧化型低密度脂蛋白(Ox-LDL)可呈浓度依赖性增加人血管平滑肌细胞(VSMCs)的钙化,FPS 处理对 Ox-LDL 诱导的人 VSMCs 钙化具有显著的保护作用。此外,FPS 处理可减轻 Ox-LDL 诱导的自噬活性下调,自噬抑制剂 3-甲基腺嘌呤可减弱 FPS 对 Ox-LDL 诱导的钙化的保护作用。综上所述,本研究首次证明,FPS 可防止 Ox-LDL 诱导的人 VSMCs 血管钙化,这可能是通过激活自噬实现的。这支持了自噬可能是一种对抗血管钙化的内源性保护机制的假说,并且 FPS 可作为血管钙化的潜在治疗药物。