本文从Tinospora crispa的藤蔓和叶子中分离出17种结构多样的克氏塔杂萜二萜类化合物，包括十种未描述的克氏塔杂萜二萜类化合物（tinopanoids K-T，1-10）和七种已知化合物（11-17）。化合物3不仅具有具有抗炎活性的γ-羟基-α，β-不饱和-γ-内酯的主要取代基，还具有C-3 / C-4的三元环氧结构。利用光谱数据解释阐明了克氏塔杂萜二萜化合物的平面结构和相对构型。化合物1、4、8和13的绝对构型由单晶X射线衍射确定，化合物3的绝对构型则是利用计算的ECD数据和相关对-溴苯甲酸酯的单晶X射线衍射确定的（3a）。随后，评估所有化合物对LPS激活的BV-2细胞产生的一氧化氮（NO）的抑制效果，化合物3和8表现出更好的NO抑制效果，IC50值分别为5.6和13.8μM，优于阳性对照米诺环素（Mino，IC50 = 22.9μM）。免疫印迹分析和qRT-PCR的对应结果表明，化合物3可以显著抑制可诱导型一氧化氮合酶（iNOS）和环氧化酶2（COX-2）的蛋白表达，以及促炎细胞因子肿瘤坏死因子-α（TNF-α）、白细胞介素6（IL-6）和白细胞介素1β（IL-1β）的mRNA水平。通过免疫印迹和免疫荧光分析，研究了化合物3发挥抗神经炎症作用的潜在机制。结果表明，化合物3通过抑制LPS诱导的神经炎症的信号转导和激活Heme Oxygenase-1 (HO-1)介导的信号途径，抑制了富含toll样受体4 (TLR4)的信号传导转录因子3（Stat3）和丝裂原激活蛋白激酶（MAPK）信号通路。© 2023 Elsevier Inc. 保留所有权利。

A total of 17 structurally diverse clerodane diterpenoids, including ten undescribed clerodane diterpenoids (tinopanoids K-T, 1-10) and seven known compounds (11-17), were isolated from the vines and leaves of Tinospora crispa. Compound 3 has not only bear the dominant substituents of γ-hydroxy-α, β-unsaturated-γ-lactone with anti-inflammatory activity, but also a ternary epoxy structure at C-3/C-4. The planar structures and relative configurations of the clerodane diterpenoids were elucidated by spectroscopic data interpretation. The absolute configurations of compounds 1, 4, 8 and 13 were determined by single-crystal X-ray crystallographic, while that of compound 3 was determined using computed ECD data and single crystal X-ray diffraction of related p-bromobenzoate ester (3a). Subsequently, all compounds were evaluated for their inhibitory effect on nitric oxide (NO) production of LPS-activated BV-2 cells, and compounds 3 and 8 exhibited better NO inhibitory potency, with IC50 values of 5.6 and 13.8 μM than the positive control minocycline (Mino, IC50 = 22.9 μM). The corresponding results of western blot analysis and qRT-PCR revealed that compound 3 can significantly inhibit the inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) protein expressions, mRNA levels of pro-inflammatory cytokins of tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6) and interleukin 1β (IL-1β). The underlying mechanism by which compound 3 exerted anti-neuroinflammatory effects was investigated by western blot and immunofluorescence assay, which suggested compound 3 inhibited LPS induced neuroinflammation via the suppression of toll-like receptor 4 (TLR4) dependent Signal Transducer and Activator of Transcription 3 (Stat3) and mitogen-activated protein kinase (MAPK) signaling pathways, and the activation of Heme Oxygenase-1 (HO-1) mediated signals.Copyright © 2023 Elsevier Inc. All rights reserved.