Evodiamine protects against airway remodelling and inflammation in asthmatic rats by modulating the HMGB1/NF-κB/TLR-4 signalling pathway

Figures & data

Figure 1. Influence of evodiamine on cytokine levels in the serum (A) and BALF (B) of asthmatic rats. Values are means ± SD (n = 8); ^@@p < 0.01 compared to the normal group; **p < 0.01 compared to the asthma group.

Table

Primer	Forward	Reverse
TLR-4	5′-CCCTGAGGCATTTAGGCAGCTA-3′	5′-AGGTAGAGAGGTGGCTTAGGCT-3′
MyD88	5′-GGCATTTCACTGCTTGATGT-3′	5′-TGACATTCCCATGAAACCTC-3′
NF-κB	5′-GCTACACAGAGGCCATTGAA-3′	5′-GTGGAGGAAGACGAGAGAGG-3′
HMGB1	5′-TTAGTCCCAGCGAAGGCTAT-3′	5′-CAAGTTTCCTGAGCAATCCA-3′
GAPDH	5′-GCAAGTTCAACGGCACAG-3′	5′-CGCCAGTAGACTCCACGAC-3′

Figure 2. Evodiamine reduces the thickness of airway smooth muscle and the airway wall in intact small bronchioles of asthmatic rats. Values are means ± SD (n = 8); ^@@ p < 0.01 compared to the normal group; ** p < 0.01 compared to the asthma group.

Figure 3. Effects of evodiamine on IFN-γ and IgE levels in lung tissue homogenates of asthmatic rats. Values are means ± SD (n = 8); ^@@p < 0.01 compared to the normal group; **p < 0.01 compared to the asthma group.

Figure 4. Effects of evodiamine on the infiltration of inflammatory cells in the BALF of asthmatic rats. Values are means ± SD (n = 8); ^@@p < 0.01 compared to the normal group; **p < 0.01 compared to the asthma group.

Figure 5. Immunohistochemical analyses of the expression of NF-κB protein in the lung tissue of asthmatic rats. Values are means ± SD (n = 8); ^@@p < 0.01 compared to the normal group; **p < 0.01 compared to the asthma group.

Figure 6. Expression of TLR-4, MyD88, NF-κB, and HMGB1 mRNA in the lung tissue of evodiamine-treated asthmatic rats. Values are means ± SD (n = 8); ^@@p < 0.01 compared to the normal group; **p < 0.01 compared to the asthma group.

Figure 7. Histopathological changes in the lung tissue of evodiamine-treated asthmatic rats. (A) H&E staining of lung tissue and histopathological score. (B) PAS staining of lung tissue and PAS score. Values are means ± SD (n = 8); ^@@p < 0.01 compared to the normal group; **p < 0.01 compared to the asthma group.

Figure 8. In silico molecular docking shows the interaction of the TLR-4 protein and evodiamine. The solid area in the protein structures represents the area of interaction.

Table 1. Docking scores for TLR-4 protein with ligand molecule Evodiamine.

Docking Score for TLR-4 with ligand Evodiamine
Mode of ligand	Affinity (kcal/mol)	Distance from rmsd l.b	Best mode rmsd u.b
1	−8.9	0.000	0.000
2	−8.8	13.120	16.496
3	−8.8	2.930	3.627
4	−8.7	1.049	2.163
5	−8.7	12.131	13.776
6	−8.6	11.944	14.646
7	−8.6	12.283	15.506
8	−8.6	11.870	14.942
9	−8.5	8.502	9.972