Wang Ruxia, Kang Zhiruo, Li Mingang, Zhang Zehui, Guo Yuru, Niu Shengli, Han Limei, Tian Chunlian, Liu Mingchun
Key Laboratory of Livestock Infectious Diseases, Ministry of Education, and Key Laboratory of Ruminant Infectious Disease Prevention and Control (East), Ministry of Agriculture and Rural Affairs, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, 120 Dongling Road Shenhe Dist., Shenyang, 110866, PR China.
College of Laboratory Animal Medicine, Liaoning University of Traditional Chinese Medicine, 79 Chongshan Road Huanggu Dist., Shenyang, 110847, PR China.
Poult Sci. 2025 Jun 7;104(9):105409. doi: 10.1016/j.psj.2025.105409.
The clinical utility of ceftiofur primarily stems from its broad-spectrum antibacterial properties. However, its limited oral bioavailability significantly constrains therapeutic effectiveness. Although natural flavonoids have been well-documented to facilitate intestinal drug transport, the potential of luteolin to enhance ceftiofur intestinal absorption and the mechanistic basis for this effect remain to be elucidated.
This study aims to investigate the effects of luteolin on duck intestinal absorption of ceftiofur and elucidate its molecular mechanisms.
This study employed duck everted gut sacs and Caco-2 cell monolayers to examine ceftiofur intestinal absorption in the absence and presence of luteolin. The specific inhibitors verapamil (P-glycoprotein, P-gp), Ko143 (breast cancer resistance protein, BCRP), and probenecid (multidrug resistance-associated protein 2, MRP2) were used to identify absorption-limiting transporters. The real-time quantitative PCR (qRT-PCR) and Western blot were performed to assess the transcription and translation of efflux transporters and tight junction proteins. Molecular docking evaluated the binding affinity of luteolin toward key transporters.
Ceftiofur was poorly absorbed in the duck intestine via an active transport mechanism. Critically, luteolin enhanced the intestinal absorption of ceftiofur. Furthermore, probenecid, the MRP2 inhibitor, promoted ceftiofur intestinal absorption. The qRT-PCR and Western blot detection results revealed that luteolin downregulated ceftiofur-related efflux transporter MRP2 and tight junction proteins (Occludin, Claudin-1, and ZO-1). Molecular docking results indicated that luteolin exhibited strong binding affinity to these key transporter proteins.
Luteolin increased ceftiofur intestinal absorption by downregulating the transcription and translation level of MRP2 or tight junction proteins (Occludin, Claudin-1, and ZO-1), supporting its role as an oral bioavailability enhancer.
头孢噻呋的临床效用主要源于其广谱抗菌特性。然而，其口服生物利用度有限，这显著限制了治疗效果。尽管天然黄酮类化合物已被充分证明可促进肠道药物转运，但木犀草素增强头孢噻呋肠道吸收的潜力及其作用机制仍有待阐明。
本研究旨在探讨木犀草素对鸭肠道头孢噻呋吸收的影响，并阐明其分子机制。
本研究采用鸭外翻肠囊和Caco-2细胞单层模型，在有无木犀草素的情况下检测头孢噻呋的肠道吸收。使用特异性抑制剂维拉帕米（P-糖蛋白，P-gp）、Ko143（乳腺癌耐药蛋白，BCRP）和丙磺舒（多药耐药相关蛋白2，MRP2）来鉴定吸收限制转运体。采用实时定量PCR（qRT-PCR）和蛋白质免疫印迹法评估外排转运体和紧密连接蛋白的转录和翻译水平。分子对接评估木犀草素与关键转运体的结合亲和力。
头孢噻呋通过主动转运机制在鸭肠道中吸收较差。关键的是，木犀草素增强了头孢噻呋的肠道吸收。此外，MRP2抑制剂丙磺舒促进了头孢噻呋的肠道吸收。qRT-PCR和蛋白质免疫印迹检测结果显示，木犀草素下调了与头孢噻呋相关的外排转运体MRP2以及紧密连接蛋白（闭合蛋白、Claudin-1和闭锁小带蛋白-1）。分子对接结果表明，木犀草素与这些关键转运蛋白具有很强的结合亲和力。
木犀草素通过下调MRP2或紧密连接蛋白（闭合蛋白、Claudin-1和闭锁小带蛋白-1）的转录和翻译水平增加了头孢噻呋的肠道吸收，支持其作为口服生物利用度增强剂的作用。
