基于纳米技术研究葶苈子炭治疗急性肺损伤的物质基础及其机制

Material basis and mechanism of Descurainiae Semen Carbonisatum in treatment of acute lung injury based on nanotechnology

目的在葶苈子炭Descurainiae Semen Carbonisatum(DSC)中发现并分离出新型纳米类成分,并利用脂多糖致大鼠急性肺损伤(acute lung injury,ALI)模型来研究其保护作用及其可能机制。方法利用马弗炉高温炭化葶苈子,并对炭化产物进行提取分离和透析得到一种水溶性的纳米类成分,命名为葶苈子炭纳米类成分(DSC nano-component,DSC-NCs)。采用透射电子显微镜(TEM)、X射线光电子能谱(XPS)等对DSC-NCs进行表征,并利用CCK-8实验来评价DSC-NCs的安全性。同时,利用脂多糖诱导ALI大鼠模型来评价DSC-NCs的保护作用,通过观察肺组织病理学形态,测定血清中细胞因子白细胞介素-1β(interleukin-1β,IL-1β)、IL-6、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量的变化以及肺组织中氧化损伤超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛的变化情况,研究DSC-NCs对大鼠ALI的保护作用及其可能机制。结果 TEM结果显示DSC-NCs外观形貌近球形,分散度良好,主要集中在2.5-6.5 nm,晶格间距为0.224nm。XPS分析显示DSC-NCs中含有C、O、N等元素,其中C、O元素占主导地位,结合红外光谱可以发现其表面官能团丰富,含有大量的羟基、羧基等基团。此外,细胞毒性实验显示DSC-NCs的安全剂量在500μg/mL以内。动物实验结果显示,DSC-NCs治疗组的肺组织破坏程度和炎性细胞浸润状态得到有效缓解,DSC-NCs能明显降低大鼠血清中TNF-α、IL-6、IL-1β的水平,升高SOD活性和减少丙二醛含量。结论 DSC-NCs对脂多糖诱导的大鼠ALI具有一定的保护作用,初步探讨其作用机制与抑制炎症细胞因子的分泌和减轻过度的氧化应激水平相关。这不仅为探索炭药的物质基础提供了新的见解,也为ALI疾病治疗药物的研究开发提供了新的途径。

Objective To discover and isolate a new nano-components in Descurainiae Semen Carbonisatum(DSC-NCs), and evaluate its protective effect and its possible mechanism through lipopolysaccharide(LPS)-induced acute lung injury(ALI) model.Methods A muffle furnace was used to carbonize Tinglizi(Descurainiae Semen), and the carbonized product was extracted, separated and dialyzed to obtain a water-soluble nano-component, named Descurainiae Semen Carbonisatum nano-components(DSC-NCs).Modern instruments such as transmission electron microscope(TEM) and X-ray photoelectron spectroscopy(XPS) were used to characterize DSC-NCs, and CCK-8 experiments were used to evaluate the safety of DSC-NCs. At the same time, the LPS-induced ALI rat model was used to evaluate the protective effect of DSC-NCs. The pathological morphology of lung tissue and the cytokine content changes of interleukin-1β(IL-1β), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α) in serum and superoxide dismutase(SOD)and malondialdehyde(MDA) in oxidative damage of lung tissue were observed to study the protective effect of DSC-NCs on ALI in rats and its possible mechanism. Results TEM results showed that the DSC-NCs had a nearly spherical appearance and good dispersion, mainly concentrated in the range of 2.5—6.5 nm, with a lattice spacing of 0.224 nm. XPS analysis showed that DSC-NCs contained C, O, N and other elements, of which C and O were the dominant elements. Combined with infrared spectroscopy, it could be found that the surface functional groups were rich, containing a large number of hydroxyl, carboxyl and other groups. In addition,cytotoxicity experiments showed that the safe dose of DSC-NCs was within 500 μg/mL. The results of animal experiments showed that the degree of lung tissue destruction and inflammatory cell infiltration in the DSC-NCs treatment group were effectively alleviated.DSC-NCs could significantly reduce the levels of TNF-α, IL-6, and IL-1β in rat serum, increase the activities of SOD and reduce the content of MDA. Conclusion DSC-NCs had a certain protective effect on LPS-induced ALI in rats. The preliminary study of its mechanism was related to inhibiting the secretion of inflammatory cytokines and reducing excessive oxidative stress. This research not only provides new insights for exploring the material basis of charcoal drugs, but also provides a new way for the research and development of therapeutic drugs for ALI diseases.