Research Center of Natural Resources of Chinese Medicinal Materials and Ethnic Medicine, Jiangxi University of Chinese Medicine, Nanchang, 330004, China.
Research Center of Natural Resources of Chinese Medicinal Materials and Ethnic Medicine, Jiangxi University of Chinese Medicine, Nanchang, 330004, China.
J Ethnopharmacol. 2024 Nov 15;334:118515. doi: 10.1016/j.jep.2024.118515. Epub 2024 Jul 6.
In Mongolian medicine, Loulu flower (LLF), the dried inflorescence of Rhaponticum uniflorum (L.) DC. from the Compositae family, has been used to clear heat and relieve toxicity for millennia, particularly in the treatment of pneumonia.
To reveal the effects of LLF on mice with lipopolysaccharide (LPS)-stimulated acute lung injury (ALI) and elucidate the underlying mechanisms.
ALI was established in BALB/c mice via nasal drops administration of LPS (5 mg/kg). The mice were then orally administrated with various doses of LLF extracts and the positive drug dexamethasone (DEX, 5 mg/kg), once daily for seven consecutive days. Last day, after being stimulated with LPS for 6h, the mice were closed dislocation of cervical vertebra, the serum, bronchus alveolar lavage fluid (BALF) and lung tissue were put into the EP tube and stored at -80 °C for further analysis. The changes of histopathology were tested by hematoxylin and eosin stain (H&E), the levels of, IL-1β, IL-18, TNF-α and IL-4 in BALF and serum were measured by ELISA. The pathways related to the treatment of ALI were predicted by network pharmacology. The expression levels of TLR4/NF-κB and NLRP3 signaling pathway-associated proteins, COX-2 and ERK were tested by western blotting. The levels of P65 and NLRP3 in lung tissues were determined by immunofluorescence analysis.
LLF total extract and the extract parts could alleviate the inflammatory cell infiltration, thicken the alveolar walls in lung tissues, reduce the levels of IL-18, IL-1β in BALF, the TNF-α in both BALF and serum, meantime enhance the level of IL-4 in BALF and serum in mice with LPS-induced ALI. Our network pharmacology and comprehensive gene ontology analyses revealed the active constituents of LLF and the pathways, including TLR4/NF-κB, NLRP3 and MAPK signaling pathways, which play significant roles in ALI. Furthermore, both the total extract and its extraction portions suppressed the expressions of proteins related with the COX-2, p-ERK and TLR4/NF-κB signaling pathway (TLR4, p-IκB, p-p65), as well as the NLRP3 signaling pathway (NLRP3, cleaved caspase-1, caspase-1, IL-1β).
LLF could improve the pathological changes and reducing inflammatory reactions in mice induced by LPS. The mechanism may be related to the modulation of the TLR4/NLRP3 signaling pathways.
在蒙医中，柳兰花（LLF），即菊科柳兰属植物柳兰的干燥花序，已被用于清热解毒数千年，特别是在治疗肺炎方面。
揭示 LLF 对脂多糖（LPS）刺激的急性肺损伤（ALI）小鼠的作用，并阐明其潜在机制。
通过鼻腔滴注 LPS（5mg/kg）在 BALB/c 小鼠中建立 ALI。然后，将小鼠口服不同剂量的 LLF 提取物和阳性药物地塞米松（DEX，5mg/kg），每天一次，连续七天。最后一天，用 LPS 刺激 6h 后，颈椎脱位处死小鼠，将血清、支气管肺泡灌洗液（BALF）和肺组织放入 EP 管中，于-80°C保存，以备进一步分析。通过苏木精和伊红染色（H&E）检测组织病理学变化，通过 ELISA 法检测 BALF 和血清中 IL-1β、IL-18、TNF-α和 IL-4 的水平。通过网络药理学预测与 ALI 治疗相关的途径。通过 Western blot 检测 TLR4/NF-κB 和 NLRP3 信号通路相关蛋白 COX-2 和 ERK 的表达水平。通过免疫荧光分析测定肺组织中 P65 和 NLRP3 的水平。
LLF 总提取物及其提取物部分可减轻 LPS 诱导的 ALI 小鼠肺组织中炎性细胞浸润，肺泡壁增厚，降低 BALF 中 IL-18、IL-1β水平，降低 BALF 和血清中 TNF-α水平，同时提高 BALF 和血清中 IL-4 水平。我们的网络药理学和综合基因本体分析揭示了 LLF 的活性成分以及 TLR4/NF-κB、NLRP3 和 MAPK 信号通路等在 ALI 中起重要作用的途径。此外，总提取物及其提取部分均抑制了与 COX-2、p-ERK 和 TLR4/NF-κB 信号通路（TLR4、p-IκB、p-p65）以及 NLRP3 信号通路（NLRP3、cleaved caspase-1、caspase-1、IL-1β）相关的蛋白表达。
LLF 可改善 LPS 诱导的小鼠的病理变化和减轻炎症反应。其机制可能与 TLR4/NLRP3 信号通路的调节有关。
