Li Qingzhong, Yang Shurong, Yang Shuling, Xin Fuzhen, Wang Meijing
Department of Anesthesia, the Third Hospital of Jinan, Jinan 250132, PR China.
Clinical Laboratory of Jinan municipal Central Hospital, Jinan 250014, PR China.
Int Immunopharmacol. 2015 Sep;28(1):724-30. doi: 10.1016/j.intimp.2015.07.035. Epub 2015 Aug 7.
This study was designed to investigate the anti-inflammatory effect of phlomisoside F (PMF) isolated from Phlomis younghusbandii and to explore the possible pharmacological mechanisms. Anti-inflammatory effects of PMF were evaluated by using carrageenan-induced rat paw edema test, dimethylbenzen-induced ear edema test, acetic acid-induced vascular permeability and cotton pellet granuloma test. Furthermore, the releases of pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) were determined by ELISA. To explore the potential mechanisms, expressions of iNOS and COX-2 were determined by quantitative real-time PCR and western blotting assays. In addition, the expressions of nuclear p65, cytosolic p65, IκB, p38, p-p38, p-ERK1/2, ERK1/2, JNK and p-JNK were determined by western blotting assay. Our results indicated that PMF administered orally could not only significantly decrease rat paw edema in rats and ear edema in mice, but also reduce the vascular permeability in mice and granuloma weights in rats. In vitro, the releases of LPS-induced pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) and enzymes (iNOS and COX-2) were decreased significantly by PMF treatment in RAW 264.7 cells. In addition, the LPS-induced up-regulations of nuclear p65, p38, p-p38, p-ERK1/2, JNK and p-JNK proteins in RAW 264.7 cells significantly decreased by PMF, and expressions of cytosolic p65 and IκB were obviously up-regulated after treatment with PMF. In conclusion, we suggested that the PMF is a promising potential anti-inflammatory drug, and PMF could down-regulate expressions of pro-inflammatory cytokines and mediators by inhibiting the NF-κB/MAPK pathways.
本研究旨在探讨从绵参中分离得到的绵参苷F（PMF）的抗炎作用，并探索其可能的药理机制。通过角叉菜胶诱导的大鼠足跖肿胀试验、二甲苯诱导的小鼠耳肿胀试验、醋酸诱导的血管通透性试验和棉球肉芽肿试验评估PMF的抗炎作用。此外，采用酶联免疫吸附测定法（ELISA）检测促炎细胞因子（TNF-α、IL-6和IL-1β）的释放。为探索潜在机制，通过定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测诱导型一氧化氮合酶（iNOS）和环氧化酶-2（COX-2）的表达。另外，采用蛋白质免疫印迹法检测细胞核p65、细胞质p65、IκB、p38、磷酸化p38（p-p38）、磷酸化细胞外信号调节激酶1/2（p-ERK1/2）、细胞外信号调节激酶1/2（ERK1/2）、c-Jun氨基末端激酶（JNK）和磷酸化JNK（p-JNK）的表达。我们的结果表明，口服PMF不仅能显著减轻大鼠足跖肿胀和小鼠耳肿胀，还能降低小鼠血管通透性和大鼠肉芽肿重量。在体外，PMF处理可显著降低RAW 264.7细胞中脂多糖（LPS）诱导的促炎细胞因子（TNF-α、IL-6、IL-1β）和酶（iNOS和COX-2）的释放。此外，PMF可显著降低LPS诱导的RAW 264.7细胞中细胞核p65、p38、p-p38、p-ERK1/2、JNK和p-JNK蛋白的上调，并且PMF处理后细胞质p65和IκB的表达明显上调。总之，我们认为PMF是一种有前景的潜在抗炎药物，并且PMF可通过抑制核因子κB（NF-κB）/丝裂原活化蛋白激酶（MAPK）通路下调促炎细胞因子和介质的表达。
