Figure 1.
Effect of methotrexate and/or BV on ankle diameter, paw volume and paw withdrawal latency.
Panel A: percentage change in ankle diameter, panel B: paw volume changes and panel C: percentage inhibition of paw withdrawal latency (PWL) from pre-injection values in adjuvant induced arthritic rats over a period of 21 days. Data are represented as mean ± SD. a or b: significantly different from the corresponding Arth or MTX group, respectively at P<0.05 using repeated measures of ANOVA followed by Tukey-Kramer Multiple Comparison Test.
Figure 2.
Effect of methotrexate and/or BV on arthritic index and gait score.
Panel A: arthritic index score and panel B: gait score on day 21 in adjuvant induced arthritis. Data are represented as medians and interquartile ranges (n = 10). a or b: significantly different from the corresponding Arth or MTX group, respectively at P<0.05 using Kruskal-Wallis test followed by Dunn's Multiple Comparison Test.
Figure 3.
Methotrexate and/or BV effect on Tissue necrosis factor-alpha expression in synovial membrane of hind paw.
Immunohistochemical staining (×100) of paw sections of A: normal rat shows almost negative immunostaining for TNF-α, B: arthritic non-treated rat shows massive immunostaining, C: MTX treated rat shows moderate immunostaining, D: BV treated rat shows mild immunostaining and E: concurrently treated rat with MTX and BV shows minimal TNF-α expression. F: mean optical density of synovial membrane stained with TNF-α immunostaining in different studied groups. Data are represented as mean ± SD (n = 10). a, b, c or d: significantly different from the corresponding Normal, Arth, MTX or BV group respectively at P<0.05 using one-way ANOVA followed by Tukey-Kramer Multiple comparison test.
Figure 4.
Methotrexate and/or BV effect on NF-κB (p65) expression in synovial membrane of hind paw.
Immunohistochemical staining (×400) of paw sections of A: normal rat shows almost negative immunostaining, B: arthritic non-treated rat shows massive immunostaining, C: MTX treated rat shows moderate immunostaining, D: BV treated rat shows mild immunostaining, E: concurrently treated rats with MTX and BV shows minimal NF-κB p65 expression. F: mean optical density of synovial membrane stained with NF-κB p65 immunostaining in different studied groups. Data are represented as mean ± SD (n = 10). a, b, c or d: significantly different from the corresponding Normal, Arth, MTX or BV group respectively at P<0.05 using one-way ANOVA followed by Tukey-Kramer Multiple comparison test.
Figure 5.
Photomicrographs of hind paw sections stained by H&E (×100).
A: paw section from normal rat shows normal appearance of synovial membrane (syn), cartilage (cart) and bone. B: paw section from arthritic non-treated rat shows extensively expanding synovial membrane with pannus formation. C: paw section from arthritic rat treated with MTX shows moderate infiltration with pannus formation. D: paw section from arthritic rat treated with BV alone shows normal synovial membrane with mild infiltration. E: paw section from arthritic rat concurrently treated with MTX and BV shows intact histological structure of cartilage, bone and synovial membrane.
Table 1.
Methotrexate and/or bee venom effect on serum AST, ALT and TNF-α in adjuvant arthritic rats.
Figure 6.
Methotrexate and/or BV effect on NF-κB (p65) expression in liver.
Immunohistochemical staining (×400) of liver sections of A: normal rat shows almost negative immunostaining, B: arthritic non-treated rat shows moderate immunostaining, C: MTX treated rat shows intense immunostaining, D: BV treated rat shows mild immunostaining and E: concurrently treated rat with MTX and BV shows minimal NF-κB p65 expression. F: mean optical density of liver tissue sections immunostained with NF-κB p65 in different studied groups. Data are represented as mean ± SD (n = 10). a, b, c or d: significantly different from the corresponding Normal, Arth, MTX or BV group respectively at P<0.05 using one-way ANOVA followed by Tukey-Kramer Multiple comparison test.
Figure 7.
Photomicrographs of liver sections stained by H & E (×400).
A: liver section of normal rat shows normal central vein (CV) and surrounding hepatocytes (h). B: liver section of arthritic non-treated rat shows degeneration in hepatocytes (d). C and D: liver sections of arthritic rat treated with MTX alone shows severe fatty change (f), degeneration (d) of hepatocytes, congested portal tract (PT) and lost cell boundaries with distortion of normal architecture (circle). E: liver section of arthritic rat treated with BV alone shows normal hepatic architecture. F: liver section of arthritic rat concurrently treated with MTX and BV showing mild congestion in the central vein (CV) with diffuse kupffer cells proliferation (arrow) in between hepatocytes.
Figure 8.
Bee venom effect on plasma concentrations, synovial fluid and tissue concentrations of methotrexate single dose.
Panel A: plasma concentrations of methotrexate, panel B: different tissue concentrations of methotrexate. Data are represented as mean ± SD or percentage change of MTX group (n = 15).* means significantly different from the corresponding MTX group at P<0.05 using unpaired student's t-test.
Table 2.
Effect of bee venom on pharmacokinetic parameters and constants of a single dose of methotrexate.