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. 2020 May 25;11:760. doi: 10.3389/fphar.2020.00760

Figure 3.

Figure 3

Circumdatin D downregulated inflammatory cytokines in LPS-induced BV-2 cells. (A) Cells were stimulated with 1 μg/ml LPS in the absence or presence of circumdatin D (10, 20, and 50 μM) for 6 h for measurement of TNF-α, and 8 h for measurement of IL-1β. The production of TNF-α and IL-1ß were measured by ELISA. (B) Cells were stimulated with 1 μg/ml LPS in the absence or presence of circumdatin D (10, 20, and 50 μM) for 24 h. Total RNA was prepared, and the mRNA levels of TNF-α, IL-1β, iNOS and COX-2 were determined using RT-PCR. GAPDH mRNA served as a control. (C) Cells were stimulated with 1 μg/ml LPS in the absence or presence of circumdatin D (10, 20, and 50 μM) for 24h. The protein expressions of iNOS and COX-2 were determined by Western blot assay. The data are represented as a mean ± S.D. from independent experiments performed in triplicate (#compared with the control, *compared with LPS, */#P < 0.05, **/##P < 0.01, ***P < 0.001).