69As comparison with untreated CIAA mice, pretreatment by ART (60 μg/ml)
or RAP (30 μg/ml) for three days, or posttreatment by ART (60 μg/ml) or RAP
(50 μg/ml) for five days leads to SpO 2 increase to a normal reading. These results
demonstrated that ART and RAP can mitigate NO-mediated hypoxia, either in
pretreatment or in posttreatment. After treatment of LIAA mice with 60 μg/
ml ART or 0.5 mg/ml l-NMMA, SpO 2 elevates to as high as 79.67 ± 2.08 % or
81.67 ± 2.89 %, higher than LIAA mice (67.00 ± 1.73 %), but lower than control
mice (98.33 ± 0.58 %).
5.3.2.5 Effects of ART, RAP, and/or BLA on Synovial
Angiogenesis, Glycolysis, and Inflammation
The upregulation of HIF-1α and VEGF in CIAA mice were reversed by pretreat-
ment or posttreatment with ART or RAP, in which a more effectively inhibi-
tory effect on the induction of HIF-1α and VEGF was observed in pretreatment
mice than in posttreatment mice. For example, posttreatment by RAP (50 μg/
ml) upregulates HIF-1α for 2-folds. While HIF-1α is downregulated by pretreat-
ment, VEGF remains unchanged. LIAA mice treated by ART (60 μg/ml) show the
lower expression levels of HIF-1α and VEGF than untreated LIAA mice. In simi-
lar, l-NMMA (0.5 mg/ml) also downregulates HIF-1α and VEGF. The therapeutic
effects of ART and ART + BLA on the expression of HIF-1α, VEGF, and iNOS in
LIAA and CIAA mice were listed in Table 5.3.
5.3 ART Alleviates Adjuvant/LPS-Induced Synovitis
Table 5.3 Immunohistochemical analysis of HIF-1α, VEGF, and iNOS in LIAA and CIAA
mice treated by ART or ART + BLA
ART artemisinin; BLA betulilic acid; CIAA collagen-induced acute arthritis; HIF- 1 α hypoxia-
inducible factor 1α; iNOS inducible NOS; LIAA LPS-induced acute arthritis; VEGF vascu-
lar endothelial growth factor. The singular asterisk (*) represents different from the model
(p < 0.05); Double asterisks (**) represent very different from the model (p < 0.01); The singular
pound sign (#) represents different from the control (p < 0.05); Double pound signs (##) repre-
sent very different from the control (p < 0.01)
Group Immunohistochemical staining strength (n = 3)
HIF-1α VEGF iNOS
Control 11.81 ± 1.02 9.04 ± 2.00 13.45 ± 3.12
LIAA mice treated by 60 μg/ml 13.82 ± 1.22* 20.05 ± 3.11** 51.72 ± 4.48*
LIAA mice treated by 100 μg/ml BLA 22.83 ± 1.98* 11.27 ± 3.87** 18.45 ± 1.65**
LIAA mice treated by 60 μg/ml ART 75.19 ± 3.03 60.63 ± 4.51* 36.55 ± 5.25*
LIAA mice 99.96 ± 4.71# 110.68 ± 4.55## 92.67 ± 4.75##
CIAA mice treated by 60 μg/ml 32.16 ± 2.69** 7.01 ± 3.56* 23.45 ± 5.33*
CIAA mice treated by 100 μg/ml BLA 51.24 ± 3.95* 10.67 ± 1.98 64.73 ± 1.98*
CIAA mice treated by 60 μg/ml ART 26.50 ± 5.53* 9.45 ± 4.15* 8.13 ± 1.08*
CIAA mice 100.21 ± 5.88## 19.35 ± 3.79# 156.85 ± 5.55#