School of Pharmacy, Jiangxi University of Chinese Medicine, Nanchang, China; College of Traditional Chinese Medicine, Jiangxi University of Chinese Medicine, Nanchang, China.
School of Pharmacy, Jiangxi University of Chinese Medicine, Nanchang, China.
J Ethnopharmacol. 2025 Jan 30;337(Pt 1):118807. doi: 10.1016/j.jep.2024.118807. Epub 2024 Sep 7.
The dried rattan stem of Tinospora sinensis (Lour.) Merr. is valued for its efficacy of clearing heat and removing toxicity, calming and soothing the nerves. It is widely used in Tibetan medicine for the treatment of rheumatic and aging diseases. Studies have confirmed its anti-inflammatory and ameliorating effects on Alzheimer's disease; however, the anti-neuroinflammation efficacy and mechanism remain unclear.
This study aimed to explore the anti-neuroinflammation efficacy, major effective ingredients, and potential mechanism of extract of Tinosporae sinenisis (TIS).
UPLC-Q-TOF/MS was used to identify the compounds of TIS and the plasma components of rats after gastric administration of TIS. C57BL/6 J mice were continuously intraperitoneally injected with lipopolysaccharide (LPS) (250 μg/kg) for 14 d to establish a neuroinflammation model. The effects of TIS (4.5 g/kg, 9 g/kg) on the learning and memory abilities in mice with neuroinflammation was evaluated using spontaneous activity, novel object recognition, and Morris water maze tests. Pathological changes in the hippocampus were observed using hematoxylin and eosin staining. Gene and protein levels of inflammatory factors in the brain were detected using qRT-PCR and ELISA kits. Iba-1 levels in the brain were detected using immunofluorescence to assess the degree of microglial activation. Network pharmacology, based on the components absorbed into plasma of TIS, was used to predict potential targets and pathways. Proteomics was used to study the differentially expressed proteins and related pathways in the brain tissue of mice with neuroinflammation. Finally, correlation analysis was performed on the results of network pharmacology and proteomics, and proteins related the anti-neuroinflammatory effect of TIS were detected by western blot.
A total of 39 compounds were identified in TIS: genipingentiobioside, isocorydin, reticuline, (-)-argemonine, tinosineside A, tinosinenside A, and costunolide were absorbed into the plasma. After continuous intraperitoneal injection of LPS into C57BL/6 J mice, microglia in the brain tissue were activated and the gene and protein levels of IL-1β, TNF-α, IL-6, and iNOS were increased in the brain tissue, suggesting that the neuroinflammation model was successfully established. TIS reduced Iba-1 levels and gene expression and protein levels of inflammatory factors in the brain of mice with neuroinflammation. Furthermore, TIS improved the pathological changes in the hippocampus and learning and memory abilities caused by neuroinflammation. Network pharmacology has predicted that TNF, IL-1β, and IκBKB are closely related to neuroinflammation. Proteomics identified key differentially expressed proteins, including TNF, NF-κB2, NF-κBIA, and TLR4. Toll-like receptor (TLR), NF-κB, and NOD-like receptor (NLR) signaling pathways are involved in neuroinflammation-related pathways. Correlation analysis revealed TLR, TNF and NLR signaling pathways were closely related to the anti-neuroinflammatory effects of TIS. We observed that TIS alleviated neuroinflammation by inhibiting the TLR4/NF-κB/NLRP3 pathway.
Thirty-nine compounds were identified from TIS, among which seven were absorbed into the plasma as prototype components. TIS alleviated LPS-induced neuroinflammation in mice, and its mechanism was related to inhibition of TLR4/NF-κB/NLRP3 signaling pathway.
三叶崖爬藤（Tinospora sinensis（Lour.）Merr.）的干燥藤茎因其清热解毒、镇静安神的功效而备受重视。它在藏药中被广泛用于治疗风湿和衰老疾病。研究证实其具有抗炎和改善阿尔茨海默病的作用；然而，其神经炎症的疗效和机制仍不清楚。
本研究旨在探讨三叶崖爬藤提取物（TIS）的抗炎作用、主要有效成分和潜在机制。
采用 UPLC-Q-TOF/MS 鉴定 TIS 的化合物和大鼠灌胃 TIS 后的血浆成分。用脂多糖（LPS）（250μg/kg）连续腹腔注射 C57BL/6J 小鼠 14 天建立神经炎症模型。采用自发活动、新物体识别和 Morris 水迷宫试验评价 TIS（4.5g/kg、9g/kg）对神经炎症小鼠学习记忆能力的影响。采用苏木精-伊红染色观察海马组织的病理变化。采用 qRT-PCR 和 ELISA 试剂盒检测脑内炎症因子的基因和蛋白水平。采用免疫荧光法检测脑内 Iba-1 水平，评估小胶质细胞激活程度。基于 TIS 吸收到血浆中的成分进行网络药理学预测潜在靶点和通路。采用蛋白质组学研究神经炎症小鼠脑组织中的差异表达蛋白和相关通路。最后，对网络药理学和蛋白质组学的结果进行相关性分析，并通过 Western blot 检测与 TIS 抗炎作用相关的蛋白质。
在 TIS 中鉴定出 39 种化合物：京尼平龙胆二糖苷、异柯因、新斯大环碱、（-）-argemonine、tinosineside A、tinosinenside A 和木香烃内酯被吸收到血浆中。连续腹腔注射 LPS 后，C57BL/6J 小鼠脑内小胶质细胞被激活，脑内 IL-1β、TNF-α、IL-6 和 iNOS 的基因和蛋白水平升高，表明成功建立了神经炎症模型。TIS 降低了神经炎症小鼠脑内 Iba-1 水平以及炎症因子的基因和蛋白表达。此外，TIS 改善了神经炎症引起的海马组织病理变化和学习记忆能力。网络药理学预测 TNF、IL-1β和 IκBKB 与神经炎症密切相关。蛋白质组学鉴定出关键差异表达蛋白，包括 TNF、NF-κB2、NF-κBIA 和 TLR4。Toll 样受体（TLR）、NF-κB 和 NOD 样受体（NLR）信号通路涉及神经炎症相关通路。相关性分析表明 TLR、TNF 和 NLR 信号通路与 TIS 的抗炎作用密切相关。我们观察到 TIS 通过抑制 TLR4/NF-κB/NLRP3 通路缓解神经炎症。
从 TIS 中鉴定出 39 种化合物，其中 7 种作为原型成分被吸收到血浆中。TIS 缓解了 LPS 诱导的小鼠神经炎症，其机制与抑制 TLR4/NF-κB/NLRP3 信号通路有关。
