Key Laboratory of Animal Diseases and Environmental Hazards of Sichuan Province, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China.
Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu 611130, China.
Nutrients. 2022 Jan 19;14(3):439. doi: 10.3390/nu14030439.
Ageratina adenophora is an invasive plant known for its toxicity to livestock. Current research on this plant has shifted from toxicity prevention to the beneficial utilization of plant resources. This study was performed to investigate the effects and mechanisms of cryptochlorogenic acid (CCGA) isolated from Ageratina adenophora on the inflammatory responses induced by lipopolysaccharide (LPS) in RAW264.7 cells. RAW264.7 cells were pretreated with CCGA (200, 100, and 50 μg/mL) and subsequently stimulated with LPS (1 μg/mL) for 16 h. The cytotoxicity of CCGA was tested using the Cell Counting Kit (CCK8). The mechanism of action of CCGA in attenuating inflammation was also identified using enzyme-linked immunosorbent assay (ELISA), quantitative reverse transcription-polymerase chain reaction, and Western blot. The results showed that CCGA had a maximal safe concentration of 200 mg/mL. Moreover, CCGA reduced the level of nitric oxide (NO) and iNOS in LPS-induced RAW264.7 cells (p < 0.01). In addition, CCGA reduced the levels of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, and IL-8) and cyclooxygenase-2 (COX-2) in LPS-induced RAW264.7 cells at both the mRNA and protein levels (p < 0.01). CCGA prevented the activation of nuclear factor-kappa B (NF-kB) in LPS-induced RAW264.7 cells via the inhibition of IKK and IκB phosphorylation and the degradation of IκB proteins (p < 0.01). This finding indicated that CCGA isolated from A. adenophora may be a potential candidate for the treatment of inflammation-related diseases.
紫茎泽兰是一种已知对牲畜有毒的入侵植物。目前对该植物的研究已从毒性预防转向植物资源的有益利用。本研究旨在探讨从紫茎泽兰中分离出的绿原酸（CCGA）对脂多糖（LPS）诱导的 RAW264.7 细胞炎症反应的影响及其作用机制。用绿原酸（200、100 和 50μg/ml）预处理 RAW264.7 细胞，然后用 LPS（1μg/ml）刺激 16 小时。用细胞计数试剂盒（CCK8）检测 CCGA 的细胞毒性。用酶联免疫吸附试验（ELISA）、定量逆转录聚合酶链反应和 Western blot 鉴定 CCGA 减轻炎症的作用机制。结果表明，CCGA 的最大安全浓度为 200mg/ml。此外，CCGA 降低了 LPS 诱导的 RAW264.7 细胞中一氧化氮（NO）和诱导型一氧化氮合酶（iNOS）的水平（p<0.01）。此外，CCGA 降低了 LPS 诱导的 RAW264.7 细胞中促炎细胞因子（TNF-α、IL-1β、IL-6 和 IL-8）和环氧化酶-2（COX-2）的水平，在 mRNA 和蛋白水平均有统计学意义（p<0.01）。CCGA 通过抑制 IKK 和 IκB 磷酸化以及 IκB 蛋白的降解，防止 LPS 诱导的 RAW264.7 细胞中核因子-κB（NF-κB）的激活（p<0.01）。这一发现表明，从紫茎泽兰中分离出的 CCGA 可能是治疗炎症相关疾病的潜在候选药物。
