Artemisinin and Nitric Oxide Mechanisms and Implications in Disease and Health

60 5 ART for Anti-inflammation

We can conveniently evaluate the therapeutic efficacy of CEF, phytol, alcohol,
or a drug combination on BIA by measuring the hypoxic parameters, including
NO, LA, and SpO 2. From these measurements, we noticed that CEF injection or
diluted alcohol drinking can eradicate the hypoxia-derived consequences, but phy-
tol only partially normalizes the hypoxic parameters (Table 5.1).
In CIA mice established by intradermal or intra-articular injection of CII-CFA,
on the other hand, administration of ART or RAP enables a dramatic decrease of
the NO level, even lower than the control. A combination of ART with RAP, or
RAP with alcohol in BIA-CIA mice leads to the considerable repression of NO
production. In particular, an extremely lower NO level (0.448 mM) was detected
in an ART-injected CIA mouse. In contrast, untreated BIA-CIA and CIA mice
exhibit higher NO levels, up to 10 mM in the maximum.
Intriguingly, CIA mice without any treatment can give rise to a low NO level
almost equal to that of the control, which might be attributed to the decay of CII-
CFA-triggered NO after a longer duration of postimmunization. Actually, three
weeks have been passed from CII-CFA boosting to NO detection.

Table 5.1 Measurement of hypoxic parameters for evaluation of potential antiarthritic drugs in
BIA mice

CEF cefotaxime (subcutaneous injection); LA lactic acid; NO nitric oxide; SpO 2 saturation per-
centages of O 2. Measurements were conducted in BIA mice after drug administration for 3 days
or 5 days. The injected volume of each drug or a drug combination is 200 μl in all groups of
treatment. The singular asterisk (*) represents statistically significant difference from the control
(p < 0.05), and double asterisks (**) indicate statistically very significant difference from the
control (p < 0.01)

Treatment NO (μM) LA (mM) SpO 2 (%)

Control 7.15 ± 0.30 7.88 ± 0.13 83.00 ± 1.83

BIA (nontreatment) 9.87 ± 0.48* 12.58 ± 0.19** 70.50 ± 2.08*

Injection of BIA with 15 μg/ml

CEF for 3 days

2.97 ± 0.13** 7.83 ± 0.07 80.50 ± 0.71

Injection of BIA with 15 μg/ml

CEF for 5 days

3.12 ± 0.15** 7.91 ± 0.05 82.50 ± 0.71

Injection of BIA with 60 μg/ml

phytol for 3 days

5.14 ± 0.13* 6.79 ± 0.05 69.50 ± 0.71*

Injection of BIA with 60 μg/ml

phytol for 5 days

4.62 ± 0.15* 11.50 ± 0.05** 78.50 ± 0.71

Injection of BIA with 15 μg/ml

CEF + 60 μg/ml phytol for 3 days

7.57 ± 0.26 14.36 ± 0.05** 75.50 ± 0.71

Injection of BIA with 15 μg/ml

CEF + 60 μg/ml phytol for 5 days

4.84 ± 0.15* 13.57 ± 0.05** 80.50 ± 0.71

Injection of BIA with 60 μg/ml

phytol and feeding with 15 %

alcohol for 3 days

7.30 ± 0.13 6.79 ± 0.05 76.50 ± 0.71

Injection of BIA with 60 μg/ml

phytol and feeding with 15 %

alcohol for 5 days

5.91 ± 0.15* 8.32 ± 0.05 76.50 ± 0.71