Fig 1.
The β-hexosaminidase and histamine release rates of RBL-2H3 cells treated with 200 ng/ml anti-DNP IgE and 500 ng/mL DNP-BSA for from 15 minutes to 60 minutes and with 2.5 μg/mL-20 μg/mL compound 48/80 for 30 minutes. n = 3.
Values indicate mean±SEM. Two-way ANOVAs followed by the Sidak test were employed compared to the control group. *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.00005 compared with the control group.
Fig 2.
RBL-2H3 cells treated with H2O, 10μg/mL Compound 48/80.
For IgE-positive groups, 200 ng/mL anti-DNP IgE was added and incubated for 24 hours, and then 500 ng/mL DNP-BSA were added at the same time as drug treatment. PBS was added to IgE-negative groups as vehicles of anti-DNP IgE and DNP-BSA. A, B, C, D: Cellular morphology of RBL-2H3 cells Magnification: × 200. E, F: β-hexosaminidase and histamine release rates of RBL-2H3 cells. n = 3. Values indicate mean±SEM. Unpaired student t-tests were employed compared to the control group. Eight guinea pigs per group were respectively injected into 0.9% NaCl, 0.75 mg/kg Compound 48/80. The 2 mg OVA was injected on Day 1, Day 3, and Day 5. On Day 21, another 2 mg OVA was injected to induce IgE-mediated allergy. G, H: Related β-hexosaminidase and histamine levels in guinea pigs. Values indicate mean±SD. Unpaired student t-tests were employed between each group. #, P < 0.05; ##, P < 0.005; ###, P < 0.0005; ####, P < 0.00005, compared to control group. *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.00005.
Fig 3.
The β-hexosaminidase and histamine release rates of RBL-2H3 treated with 3 drugs.
Cells were respectively treated with H2O, 10% QKL, 10% iodixanol, and 100 μg/ml vancomycin for 30 minutes. The 200 ng/ml anti-DNP IgE was added and incubated for 24 hours, and then 500ng/ml DNP-BSA were added at the same time as drug treatment. H2O was used as the vehicle of anti-DNP IgE and DNP-BSA. Qingkailing injection = QKL; iodixanol = Iodi; vancomycin = Van. n = 3.Values indicate mean±SEM. One-way ANOVA followed by Tukey test were employed. *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.00005.
Fig 4.
Determination of components of QKL by HPLC.
(A) UV absorption at 254 nm; (B) ELSD. Column: Hypersil ODS-2 C18 column (4.6 mm × 250 mm, 5 μm). Mobile phases: methanol (A) and 0.1% formic acid in water (B). The gradient was as follows: 0-5 min: 5% A, 95% B; 5-10 min: 25% A, 75%; 10-40 min: 60% A, 40% B; 40-50 min: 80% A, 20% B; 50-60 min: 90% A, 10% B. Solvent flow rate of 1 mL/min. Temperature: 35°C.
Fig 5.
RBL-2H3 cells treated with H2O, 10μg/mL compound 48/80, 10% QKL, 10% iodixanol, and 100 μg/ml vancomycin for 30 minutes.
For IgE-positive groups, 200 ng/ml anti-DNP IgE was added and incubated for 24 hours, and then 500 ng/ml DNP-BSA were added at the same time as drug treatment. PBS was added to IgE-negative groups as vehicles of anti-DNP IgE and DNP-BSA. Magnification: × 200.
Fig 6.
The β-hexosaminidase and histamine release rates of RBL-2H3 cells treated with 10% H2O, 10 μg/mL compound 48/80, 10% QKL, 10% iodixanol, and 100 μg/ml vancomycin.
For IgE-positive groups, 200ng/ml anti-DNP IgE was added and incubated for 24 hours, and then 500ng/ml DNP-BSA were added at the same time as drug treatment. PBS was added to IgE-negative groups as vehicles of anti-DNP IgE and DNP-BSA. n = 3. Values indicate mean±SEM. Unpaired student t-tests were employed between each group. *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.00005 compared with the control group. #, P < 0.05; ##, P < 0.005; ###, P < 0.0005; ####, P < 0.00005 compared with the control group.
Fig 7.
Related β-hexosaminidase and histamine levels in guinea pigs.
Eight guinea pigs per group were respectively injected into 0.9% NaCl, 0.2 ml QKL, 0.2 mL iodixanol, and 0.1 g/kg vancomycin. The 2 mg OVA was injected on Day 1, Day 3, and Day 5. On Day 21, another 2 mg OVA was injected to induce IgE-mediated hypersensitivity. Values indicate mean±SD. Unpaired student t-tests were employed between each group. #, P < 0.05; ##, P < 0.005; ###, P < 0.0005; ####, P < 0.00005, compared to control group. *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.00005.
Fig 8.
The concentration of IgE in the blood plasma of guinea pigs induced by OVA, compound 48, and QKL injection.
Eight guinea pigs per group were treated with 0.9% NaCl (control), 0.75 mg/kg compound 48/80, 0.2 mL/300 g Qingkailing injection, 0.2 mL/300 g iodixanaol injection, and 0.1 g/kg vancomycin for 30 minutes. The 2 mg OVA was used following animal model method. The concentrations of IgE measured by Elisa kits. Values indicate mean±SD. Unpaired student t-tests were employed. *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.00005, compared to control group.
Fig 9.
The concentration of cytokines in the blood plasma of guinea pigs induced by OVA, compound 48, and QKL injection.
Eight guinea pigs per group were treated with 0.9% NaCl (control), 0.2 ml/300 g QKL injection, and 0.75 mg/kg compound 48/80 for 30 minutes. The 2 mg OVA was used following animal model method. The concentrations of IL-4, IL-13, C3a, C5a, SC5b-9, and IL-6 were measured by ELISA kits. Values indicate mean±SD. Unpaired student t-tests were employed. *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.00005, compared to control group.
Fig 10.
The concentration of cytokines in the blood plasma of guinea pigs induced by OVA, iodixanol, vancomycin and injection.
Eight guinea pigs per group were treated with 0.9% NaCl (control), 0.2 ml/300 g iodixanol injection, and 0.1 g/kg vancomycin for 30 minutes. The 2 mg OVA was used following animal model method. The concentrations of IL-4, IL-13, C3a, C5a, SC5b-9, and IL-6 were measured by ELISA kits. Values indicate mean±SD. Unpaired student t-tests were employed. *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.00005, compared to control group.