At 24 hours after the final challenge, the mice were sacrificed, and the BALF was collected, which was separated into the supernatant and cell pellet. After 2 days, mice received OAA for 1 hour. Evans blue and DNP-HSA mixture was intravenous injected 30 minutes before the termination of experiment. (D) Mice received OAA for 1 hour SH-4-54 before the injection of compound 48/80. aair-15-214-s003.ppt (165K) GUID:?EE9EC615-24AA-4AAA-ACC9-D1B0D3E9BE29 Supplementary Fig. S2 Mch-induced pulmonary resistance. Pulmonary resistance in OVA-induced allergic airway inflammation mice were detected by forced oscillation technique. The mouse was uncovered various concentration of Mch, and respiratory resistance (A), elastance (B), and compliance (C) were measured. Each data point represents the mean standard error of the mean of Mouse monoclonal to SNAI2 3 impartial experiments. aair-15-214-s004.ppt (164K) GUID:?C58C4247-D86B-4B55-96CD-4C2D428EC34B Supplementary Fig. S3 Effect of OAA on cell viability. The cytotoxicity of OAA on (A) RBL-2H3, (B) RPMCs, and (C) mBMMCs was measured by MTT assay. Each data point represents the mean standard error of the mean of 3 impartial experiments. aair-15-214-s005.ppt (145K) GUID:?08DCA0BC-0EFA-454B-89CF-C950F02A8634 Supplementary Fig. S4 Effects of OAA on IgE-mediated passive cutaneous and compound 48/80-induced systemic anaphylaxis mice models. (A) Evans blue pigmentation in the IgE-mediated PCA mice model. The ear skin of mice (n = 5/group) was sensitized by intradermal injection of anti-DNP IgE (0.5 mg/site) for 48 hours. OAA was orally administered at doses of 2, 10, and 50 mg/kg body weight 1 hour before intravenous injection of a DNP-HSA and 4% Evans blue (1:1) mixture. After 30 minutes, the ears were collected to measure dye pigmentation. (B) The dye was extracted as described in the Materials and Methods section and detected using a spectrophotometer. (C) Serum histamine levels in the compound 48/80-induced systemic anaphylaxis mice model. Each data point represents the mean standard error of the mean of 3 impartial experiments. aair-15-214-s006.ppt (279K) GUID:?D836A0DD-975B-4EF5-ADFB-54CAE18F0B27 Supplementary Fig. S5 Comparative effects of OA and OAA on OVA-induced allergic airway inflammation. (A) Serum IgE levels and BALF pro-inflammatory cytokines such as TNF- (B) and IL-4 (C) were measured by enzyme-linked immunosorbent assay. (D) -hexosaminidase levels in BALF was measured using by spectrophotometer. Each data point SH-4-54 represents the mean standard error of the mean of 3 impartial experiments. aair-15-214-s007.ppt (164K) GUID:?9EADA492-E0EF-4C49-B347-EBEDA8B3BC1B Supplementary Fig. S6 Comparative effects of OA and OAA on mast cell degranulation and pro-inflammatory cytokine expression. (A) -hexosaminidase levels were determined using a spectrophotometer. (B) Histamine levels were determined using a fluorescence plate reader. The secretion SH-4-54 of pro-inflammatory cytokines such as TNF- (C) and IL-4 (D) was measured by enzyme-linked immunosorbent assay. Each data point represents the mean standard error of the mean of 3 impartial experiments. aair-15-214-s008.ppt (170K) GUID:?F149A309-65FE-437D-A95B-A4A88E9E6FA3 Abstract Purpose Asthma is usually a complex, heterogeneous chronic inflammatory airway disease with multiple phenotypes. There has been a great progress in managing asthma, but there are still unmet needs for developing uncontrolled asthma treatments. The present study aimed to determine the effectiveness of oleanolic acid acetate (OAA) from against allergic airway inflammation and the underlying mechanism of action with a focus on mast cells. Methods To SH-4-54 investigate the effect of OAA in allergic airway inflammation, we used the ovalbumin (OVA)-sensitized and challenged mice. To examine allergic airway inflammation associated with immune responses of mast cell activation (adzuki bean) is one of the most abundant crops in Asia and has long been used in traditional medicine in Korea, China, and Japan. The water or ethanol (EtOH) extract of has been found to downregulate adipocyte proliferation and differentiation in obesity,8 phosphorylation of the JAK2/STAT3 pathway in arthritis,9 and allergic inflammation by inhibiting calcium influx and downstream Lyn/Syk/phospholipase C (PLC) signaling in mast cells.10 extracts also reduce atopic skin inflammation by regulating Th1 and Th2 responses.11 Oleanolic acid is a bioactive triterpenoid isolated from and has been studied for its anti-allergic and anti-airway inflammatory properties.12,13,14 Oleanolic acid acetate (OAA) is an improved compound with an acetyl group added to oleanolic acid. We previously reported that OAA could suppress rheumatoid arthritis by regulating T cell immune responses.15 Based on these reports, we expected that OAA inhibits allergic inflammation involved in.