Figure 12.
(A) Effect of crude C. majus latex (S1), protein fraction (S2), and alkaloid-rich fraction (S3) on the secretion of interleukin 6 (IL-6) by RAW 264.7 cells. Consecutive numbers (1–6) on the x axis correspond to different protein concentrations in the crude latex (S1) and protein fraction (S2) applied on cells: 1 = 1.6 × 10−4 mg/mL, 2 = 4.0 × 10−4 mg/mL, 3 = 1.6 × 10−3 mg/mL, 4 = 8.0 × 10−3 mg/mL, 5 = 1.6 × 10−2 mg/mL, 6 = 8.0 × 10−2 mg/mL. In the case of alkaloid-rich fraction (S3), consecutive numbers (1–6) correspond to its different dilutions: 1 = 2 × 10−4, 2 = 5 × 10−4, 3 = 2 × 10−3, 4 = 10−2, 5 = 2 × 10−2, 6 = 10−1. Cells treated with crude latex and its fractions in all dilution ranges tested (10−1–2 × 10−4) did not secrete IL-6 in detectable amounts. LPS-treated cells (100 ng/µL) (positive control) secreted IL-6 at a level of 424.7 pg/μL. (B) Simultaneous analysis of the effect of crude C. majus latex, alkaloid-rich fraction, and protein fraction on RAW 264.7 cell viability. Among the tested samples, the strongest cytotoxicity was shown for the crude latex and the alkaloid-rich fraction in the range of 8.0 × 10−3–8.0 × 10−2 mg/mL (numbers 4–6). The protein fraction showed the lowest toxicity to RAW 264.7 cells in all analyzed concentrations.