Matthew R Desrosiers  1 Alexis Mittelman  2 Pamela J Weathers  1 Affiliations

Abstract

Artemisia annua L. and artemisinin, have been used for millennia to treat malaria. We used human liver microsomes (HLM) and rats to compare hepatic metabolism, tissue distribution, and inflammation attenuation by dried leaves of A. annua (DLA) and pure artemisinin. For HLM assays, extracts, teas, and phytochemicals from DLA were tested and IC50 values for CYP2B6 and CYP3A4 were measured. For tissue distribution studies, artemisinin or DLA was orally delivered to rats, tissues harvested at 1 h, and blood, urine and feces over 8 h; all were analyzed for artemisinin and deoxyartemisinin by GC-MS. For inflammation, rats received an intraperitoneal injection of water or lipopolysaccharide (LPS) and 70 mg/kg oral artemisinin as pure drug or DLA. Serum was collected over 8 h and analyzed by ELISA for TNF-α, IL-6, and IL-10. DLA-delivered artemisinin distributed to tissues in higher concentrations in vivo, but elimination remained mostly unchanged. This seemed to be due to inhibition of first-pass metabolism by DLA phytochemicals, as demonstrated by HLM assays of DLA extracts, teas and phytochemicals. DLA was more effective than artemisinin in males at attenuating proinflammatory cytokine production; the data were less conclusive in females. These results suggest that the oral consumption of artemisinin as DLA enhances the bioavailability and anti-inflammatory potency of artemisinin.

Keywords: Artemisia annua; artemisinin; cytochrome P450 inhibition; inflammation; tissue distribution.

Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

figure 1
Figure 1 Chemical structures of artemisinin (A) and deoxyartemisinin (B).
figure2
Figure 2 Artemisinin distribution in tissues and serum of male (A) and female (B) rats after oral delivery of artemisinin as dried leaf A. annua (DLA) or pure artemisinin. n = 3, *; p ≤ 0.05; error bars = SEM.
figure3
Figure 3 Deoxyartemisinin (deoxyAN) distribution in tissues and serum of male (A) and female (B) rats after oral delivery of artemisinin as dried leaf A. annua (DLA) or pure artemisinin. n = 3, *; p ≤ 0.05; error bars = SEM.
figure4
Figure 4 Artemisinin (AN) and deoxyartemisinin (DeoxyAN) accumulation in urine of male (A,B) and female (C,D) rats after oral delivery of artemisinin as dried leaf A. annua (DLA) or pure artemisinin. n = 8, *; p ≤ 0.05; error bars = SEM.
figure5
Figure 5 Artemisinin (A) and deoxyartemisinin (B) found in the urine of male and female rats orally gavaged with either pure artemisinin or dried leaves of A. annua (DLA). n = 8; * p ≤ 0.05; error bars = SEM.
fig 6
Figure 6 Production of proinflammatory cytokines TNF-α and IL-6 in male (A,C) and female (B,D) rats after LPS challenge and treatment with either pure artemisinin (AN) or DLA (equal artemisinin doses). n = 5–6 for experimental conditions, n = 3 for injection control; error bars = SEM; *, p ≤ 0.05 when comparing LPS+DLA to LPS Control; #, p ≤ 0.05 when comparing LPS+AN to LPS Control.

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