Suppression of lipopolysaccharide-induced COX-2 expression via p38MAPK, JNK, and C/EBPβ phosphorylation inhibition by furomagydarin A, a benzofuran glycoside from Magydaris pastinacea

Figures & data

Figure 1. Chemical structures of the isolated compounds.

Table 1. ¹H (600 MHz) and ¹³C (100 MHz) NMR data for compounds 1–2 (CD₄O, δ in ppm).

Compound 1			Compound 2
Position	δC, type	δH (J in Hz)	δC, type	δH (J in Hz)
2 3	146.2, CH 106.6, CH	7.69, d (2.4) 6.77, d (2.4)	145.5, CH 106.2, CH	7.69, d (2.4) 6.72, d (2.4)
4	123.5, C		110.0, CH	6.94, s
5	129.1, C		130.3, C
6	142.6, C		140.7, C
7	135.2, C		135.3, C
8	144.0, C		143.1, C
9	127.1, C		125.8, C
1'a	23.6, CH2	3.11, m	25.9, CH2	3.11, m
1'b		3.27, m		3.24, m
2′	35.9, CH2	2.58, m	35.0, CH2	2.68, m
3′	177.9, C		176.5, C
1′'	29.7, CH2	3.53, d (6.6)
2′'	124.9, CH	5.11, t (6.6)
3′'	132.2, C
4′'	18.1, CH3	1.83, s
5′'	25.8, CH3	1.71, s
1′''	108.0, CH	4.69, d (7.8)	106.8, CH	4.68, d (7.8)
2′''	75.7, CH	3.57, t (8.4)	74.3, CH	3.57, t (8.4)
3′''	78.1, CH	3.47, m	76.6, CH	3.47, m
4′''	71.1, CH	3.45, m	69.7, CH	3.45, m
5′''	78.6, CH	3.34, m	77.3, CH	3-34, m
6′''a	62.5, CH2	3.76, dd (5.4, 12)	61.1, CH2	3.76, dd (5.4, 12)
6′''b		3.90, dd (2.4, 12)		3.90, dd (2.4, 12)

Figure 2. Main HMBC and DQF-COSY correlations of compounds 1–2.

Figure 3. Furomagydarin A reduced COX-2 expression in LPS-stimulated RAW264.7 macrophages. (A) Cells were treated with vehicle or indicated concentrations of furomagydarin A for 30 min, followed by the treatment with LPS (100 ng/ml) for another 24 h. The COX-2 level was determined by immunoblotting. Each column represents the mean ± SEM of seven independent experiments. (B) Cells were treated with vehicle or indicated concentrations of furomagydarin B for 30 min, followed by the treatment with LPS (100 ng/ml) for another 24 h. The COX-2 level was determined by immunoblotting. Each column represents the mean ± SEM of six independent experiments. (C) Cells were treated with furomagydarin A (1–10 μM) for 30 min, followed by the treatment with LPS (100 ng/ml) for another 6 h. The extent of COX-2 mRNA was determined by an RT-qPCR assay as described in the ‘Materials and methods’ section. Each column represents the mean ± SEM of eight independent experiments. (D) Cells were transiently transfected with COX-2-luc or COX-2–3’UTR-luc and renilla-luc for 24 h. Luciferase activity was determined after treatment with LPS (100 ng/ml) for another 24 h. Data represent the mean ± SEM of eight independent experiments performed in duplicate. *P < 0.05, compared with the control group; #P < 0.05, compared with the group treated with LPS alone. (E) Cells were treated with furomagydarin A (10 μM) for 24 h. Cell viability was then determined using MTT and trypan blue exclusion assays. Data represent the mean ± SEM of four independent experiments performed in duplicate.

Figure 4. Furomagydarin A reduced LPS-induced C/EBP, p38MAPK or JNK phosphorylation in RAW264.7 macrophages. Cells were treated with furomagydarin A for 30 min, followed by the treatment with LPS (100 ng/ml) for another 30 min. The extent of C/EBP (A), p38MAPK (B) or JNK (C) phosphorylation was determined by immunoblotting. Each column represents the mean ± SEM of six independent experiments. *p < 0.05, compared with the control group; #p < 0.05, compared with the group treated with LPS alone.