Li Xiaodie, Luo Zhengqin, Hu Tingjun, Ou Deyuan, Guo Rongrong, Li Chunjie, Yang Jian, Song Xuqin
Department of Veterinary Medicine, College of Animal Science, Guizhou University, Guiyang 550025, Guizhou Province, China.
Department of Veterinary Medicine, College of Animal Science, Guizhou University, Guiyang 550025, Guizhou Province, China; School of Animal Production Technology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, Thailand.
Int Immunopharmacol. 2025 Aug 28;161:115027. doi: 10.1016/j.intimp.2025.115027. Epub 2025 Jun 7.
Lophatherum gracile Brongn., also known as Dan-Zhu-Ye, is traditionally used to clear heat and purge fire, relieve irritability and thirst, diuresis and catharsis. Our preliminary research revealed that total flavonoid extracts from Lophantherum gracile Brongn. (TFLG) can reverse the inflammatory response induced by LPS in mice, but the exact mechanism is still unclear.
The main chemical constituents of TFLG were first analyzed via liquid chromatography tandem high-resolution mass spectrometry (LC-HRMS). We then used network pharmacology to screen potential targets of TFLG to treat inflammation. Subsequently, inflammatory models were established in 3D4/2 cells and SPF Kunming mice via lipopolysaccharide. The predicted potential targets and associated signaling pathways were further validated through ELISA, western blot analysis, and qRT-PCR.
The LC-HRMS results revealed that approximately 36 compounds with contents greater than 0.1 % were identified in TFLG. Network pharmacology analysis revealed that 103 common targets of TFLG are associated with inflammation. The results of molecular docking indicated that the main ingredients of L. gracile (quercetin, kaempferol, and luteolin) can exert anti-inflammatory effects through binding with the inflammatory targets TNF, IL-6, and IL-1β. Our experimental results demonstrated that TFLG reversed the upregulation of IL-6, IL-1β, IL-12, and TNF-α and the downregulation of IL-10 in both in vitro and in vivo inflammatory models induced by LPS. The qRT-PCR results were consistent with the ELISA results above. Western blot analysis revealed that TFLG reduced the expression levels of p65, p-p65, p-IκB-α, p38, and p-p38 and increased the expression of IκB-α.
TFLG can exert an anti-inflammatory effect by inhibiting the NF-κB and MAPK signaling pathways, which aligns with the anticipated outcomes of network pharmacology. This study offers new evidence supporting the potential application of TFLG as a medication for reducing inflammation.
淡竹叶，又称丹竹叶，传统上用于清热泻火、除烦止渴、利尿通便。我们的初步研究表明，淡竹叶总黄酮提取物（TFLG）可逆转小鼠体内脂多糖诱导的炎症反应，但其确切机制仍不清楚。
首先通过液相色谱串联高分辨率质谱（LC-HRMS）分析TFLG的主要化学成分。然后利用网络药理学筛选TFLG治疗炎症的潜在靶点。随后，通过脂多糖在3D4/2细胞和SPF级昆明小鼠中建立炎症模型。通过酶联免疫吸附测定（ELISA）、蛋白质免疫印迹分析和实时荧光定量聚合酶链反应（qRT-PCR）进一步验证预测的潜在靶点和相关信号通路。
LC-HRMS结果显示，TFLG中鉴定出约36种含量大于0.1%的化合物。网络药理学分析表明，TFLG的103个共同靶点与炎症相关。分子对接结果表明，淡竹叶的主要成分（槲皮素、山奈酚和木犀草素）可通过与炎症靶点肿瘤坏死因子（TNF）、白细胞介素-6（IL-6）和白细胞介素-1β（IL-1β）结合发挥抗炎作用。我们的实验结果表明，TFLG在脂多糖诱导的体外和体内炎症模型中均逆转了IL-6、IL-1β、IL-12和TNF-α的上调以及IL-10的下调。qRT-PCR结果与上述ELISA结果一致。蛋白质免疫印迹分析显示，TFLG降低了p65、磷酸化p65（p-p65）、磷酸化IκB-α（p-IκB-α）、p38和磷酸化p38（p-p38）的表达水平，并增加了IκB-α的表达。
TFLG可通过抑制核因子κB（NF-κB）和丝裂原活化蛋白激酶（MAPK）信号通路发挥抗炎作用，这与网络药理学的预期结果一致。本研究为TFLG作为抗炎药物的潜在应用提供了新的证据。
