目的 从生地黄炭（dried Rehmanniae Radix Carbonisata，DRC）中发现并提取生地黄炭纳米类成分（dried Rehmanniae Radix Carbonisata nano-components，DRC-NCs），利用2,4,6-三硝基苯磺酸/乙醇溶液诱导的大鼠溃疡性结肠炎（ulcerative colitis，UC）模型评价其治疗作用及可能的作用机制。方法 利用马弗炉高温煅烧生地黄，并经过萃取、滤过、透析得到DRC-NCs。利用透射电子显微镜（transmission electron microscope，TEM）、X射线衍射（X-ray diffraction，XRD）技术、紫外-可见分光光谱、荧光光谱、傅里叶变换红外光谱以及X射线光电子能谱技术等对DRC-NCs进行表征。利用CCK-8细胞毒性试验评价DRC-NCs的安全性。利用大鼠UC模型，通过比较大鼠一般情况、疾病活动指数（disease activity index，DAI）、结肠组织损伤程度、胸腺指数、脾脏指数、结肠组织中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）及白细胞介素-6（interleukin-6，IL-6）、IL-10、丙二醛（malondialdehyde，MDA）的水平、髓过氧化物酶（myeloperoxidase，MPO）和超氧化物歧化酶（superoxide dismutase，SOD）的活力，以评价DRC-NCs的治疗作用及其机制。结果 DRC-NCs在TEM下分散度良好，为近球形且粒径大小均一，粒径分布集中在1.1～2.6 nm，晶格间距为0.354 nm，主要由C、N、O等元素构成，表面含有氨基、羟基、羧基等基团。DRC-NCs的细胞毒性很低，安全质量浓度在2 500.00 μg/mL以内。动物实验结果显示，DRC-NCs可以改善UC模型大鼠的一般情况，显著降低模型大鼠的DAI评分，减轻结肠损伤，升高胸腺指数和脾脏指数。DRC-NCs可以降低TNF-α及IL-6的水平、升高IL-10的水平、降低大鼠结肠组织中MDA的水平及MPO的活力、提高SOD的活力。结论 DRC-NCs具有治疗UC的作用，其作用机制可能与降低炎症因子和氧化应激水平，提高抗炎和抗氧化能力有关。这不仅为中药炭药的探索开发提供了一个独特的研究角度，也为临床应用生地黄炭治疗UC提供了实验依据。

Objective To discover and extract dried Rehmanniae Radix Carbonisata nano-components (DRC-NCs) from dried Rehmanniae Radix Carbonisata (DRC), and evaluate its therapeutic effect and possible mechanism in rats with ulcerative colitis (UC) induced by 2,4,6-trinitrobenzene sulfonic acid/ethanol solutionts. Methods The dried Rehmanniae Radix was calcined in a muffle furnace at a high temperature to obtain DRC, which was extracted, filtered and dialyzed to obtain DRC-NCs. Transmission electron microscopy, X-ray diffraction, UV-Vis spectroscopy, fluorescence spectrum, Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy were used to characterize DRC-NCs, and CCK-8 cytotoxicity assay was used to evaluate the safety of DRC-NCs. According to the rat with UC model, the general condition, disease activity index (DAI), colon tissue injury degree, thymus index, spleen index, the content of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-10, malondialdehyde (MDA), and the activity of myeloperoxidase (MPO), and superoxide dismutase (SOD) in the colon tissues of rats were compared to evaluate the therapeutic effect of DRC-NCs and its mechanism. Results DRC-NCs were well dispersed under TEM, nearly spherical appearance with uniform particle size, with a particle size distribution concentrated at 1.1—2.6 nm and a lattice spacing of 0.354 nm, mainly composed of elements such as C, N and O, and the surface contained amino, hydroxyl and carboxyl groups. DRC-NCs had low cytotoxicity and the safe mass concentration was within 2 500.00 μg/mL. Results of animal experiments showed that DRC-NCs could improve the general condition of model rats, significantly reduce DAI score, alleviate colon injury, and increase thymus index and spleen index. DRC-NCs could decrease the contents of TNF-α and IL-6, increase the content of IL-10, decrease the content of MDA and the activity of MPO in colon tissue, and increase the activity of SOD. Conclusion DRC-NCs had a certain therapeutic effect on UC, and its mechanism may be related to reducing the levels of inflammatory factors and oxidative stress, and improving the anti-inflammatory and antioxidant capacity. This research not only provides a unique research angle for the exploration and development of Chinese herbal charcoal drugs, but also provides an experimental basis for the clinical application of DRC in the treatment of UC.

R283.6

中央高校基本科研业务费专项基金资助项目（2019-JYB-TD-001）