Li Qing, Shi Xue, Huang Hong, Gao Qian, Sun Qingya, Meng Yao, Niu Lihang, Xie Chunfeng, Yang Cheng
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, 300353, People's Republic of China; High-throughput Molecular Drug Screening Centre, Tianjin International Joint Academy of Biomedicine, Tianjin, 300070, People's Republic of China.
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, 300353, People's Republic of China.
J Ethnopharmacol. 2025 Feb 11;341:119359. doi: 10.1016/j.jep.2025.119359. Epub 2025 Jan 11.
The whole plant of Laggera alata is frequently utilize to remedy inflammatory diseases including nephritis as a traditional Chinese medicine. However, its active ingredients and mechanism of action against sepsis-associated acute kidney injury (SA-AKI) are unknown.
This study aimed to identify active compounds from L. alata that inhibit renal inflammation and ameliorate SA-AKI, and to elucidate their mechanisms of action.
The chemical constituents were separated from the ethyl acetate layer of L. alata methanol extract by column chromatography over silica gel, medium-pressure liquid chromatography and semipreparative high-performance liquid chromatography. Extensive spectroscopic techniques were applied to determine the chemical structures. The anti-inflammatory efficiency was measured by analyzing the NO production in RAW 264.7 cells. The levels of IL-6, IL-1β, CCL-2 and CCL-5 mRNA were determined by qRT-PCR. Cecal ligation and puncture (CLP) surgery is a frequently applied method to establish the mouse sepsis model. Sepsis was thus induced in mice via CLP. The effect in the treatment of SA-AKI was evaluated by H&E staining and ELISA detection. Western blotting was used to evaluate the protein levels involved in ferroptosis, NF-κB and MAPK signaling pathways.
Twelve compounds were obtained from L.alata including four unreported sesquiterpenoids (1-4). Compound 5 exhibited the most significant inhibitory effect on NO production with the IC value of 6.034 μM and could restrain the mRNA expression of inflammatory factors IL-6, IL-1β, CCL-2 and CCL-5. The in vivo results demonstrated that compound 5 alleviated the renal injury by decreasing the serum IL-6, IL-1β, Cr, and BUN levels, reducing the kidney contents of Cys-C and KIM-1, and regulating the kidney levels of MDA, GSH, ferrous iron, GPX4, FTH1, and SLC7A11. Furthermore, Compound 5 also repressed the NF-κB and MAPK pathways in vitro and in vivo.
This study revealed that compound 5 could ameliorate SA-AKI through exerting its anti-inflammatory and anti-ferroptosis effects via NF-κB and MAPK pathways. The current research supported the traditional use of L.alata in the treatment of renal diseases.
作为一种传统中药，六棱菊全草常用于治疗包括肾炎在内的炎症性疾病。然而，其有效成分以及针对脓毒症相关性急性肾损伤（SA-AKI）的作用机制尚不清楚。
本研究旨在从六棱菊中鉴定出抑制肾脏炎症并改善SA-AKI的活性化合物，并阐明其作用机制。
通过硅胶柱色谱、中压液相色谱和半制备高效液相色谱从六棱菊甲醇提取物的乙酸乙酯层中分离化学成分。应用多种光谱技术确定化学结构。通过分析RAW 264.7细胞中一氧化氮（NO）的产生来测定抗炎效率。通过qRT-PCR测定白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）、趋化因子配体2（CCL-2）和趋化因子配体5（CCL-5）mRNA的水平。盲肠结扎和穿刺（CLP）手术是建立小鼠脓毒症模型常用的方法。因此，通过CLP诱导小鼠发生脓毒症。通过苏木精-伊红（H&E）染色和酶联免疫吸附测定（ELISA）检测评估对SA-AKI的治疗效果。蛋白质印迹法用于评估铁死亡、核因子-κB（NF-κB）和丝裂原活化蛋白激酶（MAPK）信号通路中相关蛋白的水平。
从六棱菊中获得了12种化合物，包括4种未报道的倍半萜类化合物（1-4）。化合物5对NO的产生表现出最显著的抑制作用，IC值为6.034 μM，并且可以抑制炎症因子IL-6、IL-1β、CCL-2和CCL-5的mRNA表达。体内实验结果表明，化合物5通过降低血清IL-6、IL-1β、肌酐（Cr）和尿素氮（BUN）水平，减少肾脏中胱抑素C（Cys-C）和肾损伤分子-1（KIM-1）的含量，并调节肾脏中丙二醛（MDA）、谷胱甘肽（GSH）、亚铁离子、谷胱甘肽过氧化物酶4（GPX4）、铁蛋白1（FTH1）和溶质载体家族7成员11（SLC7A11）的水平，减轻肾脏损伤。此外，化合物5在体外和体内均抑制NF-κB和MAPK信号通路。
本研究表明，化合物5可通过NF-κB和MAPK信号通路发挥抗炎和抗铁死亡作用，从而改善SA-AKI。目前的研究支持了六棱菊在治疗肾脏疾病方面的传统用途。
