Casticin continues to be isolated from and found out to have anti-inflammatory and anti-tumor properties. pathological changes by suppressing Th2 cytokine expression in mice with asthma. L. is a species of Verbenaceae distributed throughout China and other East Asian areas, and its fruit has been used to treat headache, gingival swelling, inflammation, and dizziness (Matsui et al., 2012). is also used to treat cancer in Sichuan, China (Matsui et al., 2009). Casticin has been isolated from or and reported to have anti-cancer effects, inhibit inflammation in LPS-induced acute lung injury in mice, and improve the cigarette smoke-induced acute lung inflammatory response in mice (Lee et al., 2015; Wang et al., 2016; Chou et al., 2018). We previously found that casticin can reduce pro-inflammatory cytokine and ICAM-1 expression by blocking the NF-B, MAPK, and PI3K/Akt pathways in IL-1-activated A549 human lung epithelial cells (Liou et al., 2014; Liou and Huang, 2017). Therefore, we speculated that casticin may improve asthma by suppressing inflammation and oxidative stress. In the current study, asthmatic mice were treated with casticin by intraperitoneal injection to evaluate whether casticin ameliorates asthma symptoms. We also investigated the effect of casticin on regulated immune function, oxidative stress, and inflammation in the murine model of asthma. Materials and Methods Animals Female BALB/c mice (6 to 8 8 weeks old, 20C25 g) were purchased from the National Laboratory Animal Center in Taiwan. All mice were housed in an air-conventional animal house at a controlled room temperature (22C24C) with a 12 h organic light/dark cycle. Drinking water and regular chow diet had been supplied = 12 each): regular (N group), healthful mice sensitized with regular saline and received 50 l DMSO (100%) by intraperitoneal shot; OVA-sensitized mice (OVA group), mice sensitized with OVA and received 50 l DMSO by intraperitoneal shot; prednisolone control (P group), mice sensitized with OVA and received 5 mg/kg prednisolone (dissolved in 50 l 100% DMSO) by intraperitoneal shot; and OVA-sensitized mice treated with 5 or 10 mg/kg casticin (dissolved in 50 l 100% DMSO) by intraperitoneal shot (CAS5 and CAS10 groupings, respectively). Open up in another window Body 1 Aftereffect of casticin on AHR and cell matters in BALF from asthmatic mice. (A) On times 1C3 and 14, mice sensitized with OVA by intraperitoneal shot (I.P.) and challenged with 2% OVA inhalation (I.H.) on times 14, 17, 20, 23, and 27. 1 hour prior to the OVA methacholine or problem inhalation, mice had been treated with I.P. prednisolone or casticin. (B) AHR (Penh beliefs) was assessed via inhalation of raising methacholine dosages (0C40 mg/ml). (C) Inflammatory cells and total cells had been assessed in BALF and (D) shown the percentage of eosinophils in BALF. The info are shown as means SEM of three indie tests (= 12). ? 0.05, ?? 0.01 set alongside the OVA control group. Mice had been divided into regular (N), OVA-sensitized mice (OVA), prednisolone control (P), and casticin treatment (CAS5 and CAS10) groupings. Evaluation of AHR Twenty-four hours following the last problem, AHR was assessed to judge airway work as referred to previously (Huang et al., 2017). All mice inhaled aerosolized methacholine (0 to 40 mg/ml) for 3 min before getting put into a chamber to record the improved pause (Penh) for dimension of AHR using whole-body plethysmograph (Buxco Consumer electronics, Troy, NY, USA). Histopathological Evaluation of Lung Tissues Mouse lung tissue had been removed and set with 10% formalin, and inserted in paraffin and sectioned (6-m-thick) for staining as referred Rabbit polyclonal to UGCGL2 to previously (Liou et al., 2016). Tissues BIIB021 manufacturer sections had been stained with hematoxylin and eosin (H&E) to see eosinophil infiltration or regular acid-Schiff (PAS; Sigma) to judge goblet cell hyperplasia. Inflammatory Cells in Bronchoalveolar Lavage Liquid Mice had been anesthetized with BIIB021 manufacturer 4% isoflurane and BIIB021 manufacturer bronchoalveolar lavage liquid (BALF) gathered as referred to previously (Kuo et al., 2017). Quickly, the mice had been incubated with an indwelling needle in to the trachea, the lungs cleaned 3 x with 1 ml regular saline, and supernatant collected to detect the known degrees of cytokines and chemokines. After a cytospin centrifugation, Giemsa stain (Sigma) was utilized to count number the cells and assess cell morphology. Furthermore, the percentages of eosinophils attained using the cell matters in 500 BALF cells. Glutathione (GSH) Assay A Glutathione Assay Package (Sigma) was utilized to research the degrees of glutathione (glutathione disulfide, decreased glutathione, and total.