中药作为中华民族传统医学的瑰宝，蕴含着丰富的生物活性成分，在现代医疗体系中具有重要地位。炮制是中药到临床应用的关键环节，其中热处理作为核心炮制手段，通过炒制、煅制、煮制等不同方式影响药材的理化性质和生物活性。在特定温度和时间条件下，中药活性分子可能会发生结构改变，也可能突破能量势垒进行动态自组装，形成具有多尺度特征的纳米聚集体。由中药分子组装形成的碳点（Carbon dots, CDs），兼具优异的纳米材料特性和天然药物优势，表现出卓越的细胞渗透性、水溶性、生物相容性和自身生物活性。围绕栀子苷和雷公藤红素碳点的构建与生物活性开展探究，本文的研究内容主要分为以下两部分。
第一部分为栀子苷衍生碳点（Geniposide -Carbon dots, Gen-CDs）的止血活性研究。中药炭药广泛应用于出血性疾病的治疗，然而其药效物质基础和作用机制尚未得到系统阐明。栀子（Gardeniae Fructus, GF）作为一味经典中药，经炭化炮制后产生显著的止血功效，临床主要用于血热妄行所致的吐衄、尿血及胃出血等出血证候。本研究发现，在高温炮制过程中，栀子中的主要活性成分栀子苷（Geniposide, Gen）通过脱水缩合、含氧官能团分解及碳芳香化等一系列复杂化学反应，最终形成具有纳米特性的Gen-CDs。通过建立小鼠断尾和肝脏划痕出血模型，我们证实Gen-CDs能显著缩短出血时间。在血热出血（Blood-heat hemorrhage, BHH）大鼠模型中，Gen-CDs治疗组表现出优异的胃黏膜保护作用，其中高剂量组Gen-CDs（H）的胃出血抑制率高达82.78%。凝血功能检测显示，Gen-CDs主要通过激活共同凝血途径发挥促凝作用。进一步的机制研究表明Gen-CDs能显著促进血小板活化和聚集，通过调节血栓素A₂（TXA₂）和前列环素（PGI₂）的平衡增强血小板功能。综上所述，本研究首次揭示Gen-CDs可能是栀子炭（Carbonized Gardeniae Fructus，CGF）止血作用的关键有效物质，其止血特性不仅为阐明 “炒炭止血”的科学内涵提供了实验依据，也为中药分子衍生CDs的形成机制研究开辟了新思路，这一发现对传统炭药的现代化研究具有重要的理论价值。
第二部分为雷公藤红素衍生碳点（Celastrol- Carbon dots，CEL-CDs）的抗肿瘤活性研究。作为中药雷公藤（Tripterygium wilfordii Hook. f.）的主要活性成分， CEL具有良好且广谱的抗肿瘤功效。然而，CEL的水溶性低、治疗窗窄、潜在毒性等问题阻碍了其临床应用。为提高雷公藤红素（Celastrol，CEL）的生物利用度并减少全身毒性，我们将CEL制备成具有石墨烯量子点（GQDs）微观结构的CEL-CDs。相比于CEL，CEL-CDs的水溶性明显改善，并且CEL-CDs对4T1细胞的细胞毒性作用显著高于CEL，能够促进4T1细胞的凋亡，抑制其生长和转移。通过系统的化学表征和分子动力学模拟，我们发现CEL-CDs在无需额外辅料的情况下即可表现出独特的pH响应性凝胶化行为。CEL-CDs表面含氧官能团的质子化作用促使分子间氢键形成，从而在酸性肿瘤微环境（Tumor Microenvironment，TME）中自发组装成无载体水凝胶（CCDs-gel）。小鼠核磁共振成像与活体荧光成像研究证实，CEL-CDs在弱酸性TME中能迅速形成CCDs-gel，并在肿瘤部位滞留超过6天，这种长效滞留特性不仅避免了药物的快速代谢，还显著降低了全身毒性，实现了药物的靶向递送和缓释。体内抗肿瘤结果显示CEL-CDs的抗肿瘤效果优于CEL，CEL-CDs（H）组的平均肿瘤重量为0.103 g，平均肿瘤体积为275.5 mm3，肿瘤抑制率高达92.9 %。抗肿瘤机制研究表明，CEL-CDs能有效下调关键磷酸化蛋白（p-PI3K、p-Akt和p-mTOR）的表达，促进凋亡蛋白cleaved caspase-3和cleaved caspase-9的表达，从而诱导4T1细胞程序性死亡。本研究不仅证实了具有pH响应自组装特性的CEL-CDs在抑制4T1乳腺癌生长方面的显著效果，更为中药活性分子衍生CDs在生物医学领域的多功能应用开辟了新途径。

As a treasure of traditional Chinese medicine (TCM) in the Chinese nation, herbal medicine contains abundant bioactive molecules and holds a significant position in modern healthcare systems. Processing (Paozhi) is a crucial step in preparing TCM for clinical application, with heat treatment serving as the core method. Techniques such as stir-frying, calcining, and boiling alter the physicochemical properties and bioactivity of medicinal materials. Under specific temperature and time conditions, Chinese herbal molecules may undergo structural modifications or overcome energy barriers to dynamically self-assemble, forming nanoaggregates with multi-scale characteristics. Carbon dots (CDs) derived from Chinese herbal molecules self-assembly combine the excellent properties of nanomaterials with the advantages of natural medicines, demonstrating outstanding cell permeability, water solubility, biocompatibility, and intrinsic bioactivity. Focusing on the construction and bioactivity of CDs derived from geniposide and celastrol, the content of this study is mainly divided into the following two parts.
The first part is the study of hemostatic activity of carbon dots derived from Geniposide (Gen-CDs). Carbonic herbs have always been used in the treatment of hemorrhagic diseases, but their effective components and hemostatic mechanism have not been fully studied. Gardeniae Fructus (GF) is a classic Chinese herb that is charcoaled and added with a hemostatic effect. It is widely used as a treatment for blood pressure caused by bleeding and other symptoms associated with bleeding. Geniposide (Gen) is the main active molecule in GF. During high temperature processing of GF, Gen-CDs were formed by dehydration condensation, decomposition of oxygen-containing functional groups, and carbon aromatization. Experiments on the mouse tail amputation and liver injury model showed that Gen-CDs could significantly reduce the hemostasis time. The rat blood-heat hemorrhage (BHH) verified that the Gen-CDs treatment group significantly reduced gastric mucosal hemorrhagic sites in rats. The gastric bleeding inhibition rate in the Gen-CDs (H) group was 82.78%. The results of the four coagulation items showed that Gen-CDs activated the co-coagulation pathway to accelerate hemostasis. Gen-CDs can effectively promote platelet activation and aggregation. Based on coagulation factor and routine blood tests, Gen-CDs were found to induce the synthesis of platelet activating factor TXA₂ and inhibit the production of PGI₂. In summary, Gen-CDs was the key ingredient in the hemostasis of carbonized Gardeniae Fructus (CGF). Gen-CDs with rapid hemostatic properties provide a new perspective for elucidation of the material basis of carbonic herbs and the formation mechanism of CDs derived from Chinese herbal molecules.
The second part focuses on the antitumor activity of carbon dots derived from Celastrol (CEL-CDs). As a main active ingredient of Tripterygium wilfordii Hook. f., Celastrol (CEL) has shown to be effective against a wide variety of tumor types. However, the low water solubility of CEL, the narrow therapeutic window, potential toxicity, as well as other issues prevent its clinical application. To improve the bioavailability of CEL and reduce systemic toxicity, we prepared CEL-CDs, which exhibit a microstructure similar to graphene quantum dots (GQDs). CEL-CDs exhibit improved water solubility compared to CEL, as well as a markedly higher cytotoxicity against 4T1 cells, promoting apoptosis and inhibiting growth and metastasis. Chemical characterization and molecular dynamics (MD) simulations confirmed that CEL-CDs exhibit unique pH-responsive gelation behavior without additional auxiliary substances. The protonation of oxygen-containing functional groups on the surface of CEL-CDs facilitates intermolecular hydrogen bonding in the acidic tumor microenvironment (TME), resulting in the formation of carrier-free hydrogels (CCDs-gel). Magnetic resonance imaging (MRI) and fluorescence imaging in mice indicated that CEL-CDs rapidly form CCD-gel in slightly acidic TMEs and remain at the tumor site for over six days. This avoided rapid metabolism, reduced systemic toxicity, and enabled localized delivery and sustained release in tumor-bearing mice. According to antitumor results in vivo, CEL-CDs have superior antitumor efficacy to CELs. The average tumor weight in the CEL-CDs (H) group was 0.103 g, with an average tumor volume of 275.5 mm³and a tumor inhibition rate as high as 92.9%. More importantly, CEL-CDs significantly downregulated the expression levels of key phosphorylated proteins (p-PI3K, p-Akt, and p-mTOR) and activated the expression of apoptosis proteins cleaved caspase-3 and cleaved caspase-9. In conclusion, CEL-CDs with pH-responsive self-assembly properties can effectively inhibit the rapid growth of tumors in 4T1 breast cancer mouse models, laying the foundation for the multifunctional biomedical applications of natural molecule-derived CDs.