Graduate School of Biotechnology, and College of Life Science, Kyung Hee University, Yongin-si, 17104, Gyeonggi-do, Republic of Korea.
Department of Science in Korean Medicine and Brain Korea 21 Plus Program, Graduate School, Kyung Hee University, Seoul, 02447, Republic of Korea.
Int J Nanomedicine. 2022 Jan 29;17:477-494. doi: 10.2147/IJN.S338334. eCollection 2022.
Despite great advances in the field of immunotherapy, there is still a need for novel and effective immunostimulants to overcome challenges, such as instability and autoinflammatory toxicity, associated with conventional immunostimulants. Nanotechnology provides the possibility to overcome these challenges. The well-known classical Chinese formula, Hwanglyeonhaedok-tang (HHT) has been widely used to treat immune-related diseases in clinical practice.
We developed novel gold nanoparticles (AuNPs) utilizing one-pot synthesis with the herbal formula-HHT. The optimal conditions for HHT-AuNP biosynthesis were established, and physicochemical properties of the optimized HHT-AuNPs were identified using various spectrometric and microscopic techniques. Bio-TEM analysis revealed that HHT-AuNPs were highly engulfed within RAW264.7 cells without inducing cytotoxicity. The effect of HHT-AuNPs on immunostimulatory activity was evaluated in innate and adaptive immune cells (RAW264.7 macrophages and ICR mice splenocytes) using qRT-PCR, immunoblotting, and ELISA.
The HHT-AuNPs remarkably increased the nitric oxide (NO) and immune-related cytokines production by activating the mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) signaling pathways in RAW264.7 cells. Furthermore, HHT-AuNPs exerted immunostimulatory effects on mouse splenocytes by priming T/B-cells and macrophages.
The present study is the first to demonstrate that HHT-AuNPs could be utilized as immunostimulators to activate both innate and adaptive immune systems. These results provide a foundation for the application of traditional Chinese medicinal formulae in the field of nanomedicine.
尽管免疫疗法领域取得了巨大进展，但仍需要新型有效的免疫刺激剂来克服与传统免疫刺激剂相关的挑战，如不稳定性和自身炎症毒性。纳米技术提供了克服这些挑战的可能性。著名的经典中药方剂黄莲饮（Hwanglyeonhaedok-tang，HHT）已在临床实践中广泛用于治疗免疫相关疾病。
我们利用草药方剂 HHT 一锅合成法开发了新型金纳米粒子（AuNPs）。确定了 HHT-AuNP 生物合成的最佳条件，并使用各种光谱和显微镜技术鉴定了优化的 HHT-AuNPs 的物理化学性质。生物-TEM 分析表明，HHT-AuNPs 被 RAW264.7 细胞大量吞噬，而没有诱导细胞毒性。使用 qRT-PCR、免疫印迹和 ELISA 评估 HHT-AuNPs 在固有和适应性免疫细胞（RAW264.7 巨噬细胞和 ICR 小鼠脾细胞）中的免疫刺激活性。
HHT-AuNPs 通过激活 RAW264.7 细胞中的丝裂原活化蛋白激酶（MAPK）和核因子-κB（NF-κB）信号通路，显著增加了一氧化氮（NO）和免疫相关细胞因子的产生。此外，HHT-AuNPs 通过激活 T/B 细胞和巨噬细胞对小鼠脾细胞发挥免疫刺激作用。
本研究首次表明，HHT-AuNPs 可用作免疫刺激剂来激活固有和适应性免疫系统。这些结果为传统中药方剂在纳米医学领域的应用提供了基础。
