RESEARCH ARTICLE Direct Stimulatory Effects of Oral Dihydroartemisinin on Red and White Blood Cell Stem Cells in the

Lungs of Rats
Utoh-Nedosa U A1*, Akah P A2, Nedosa K S3, Onyedibe I K4, Nedosa I V5, Agbata C A1, Okoye T C2, Njoku G6
Abstracts: Dihydroartemisinin (DHA) has been shown to produce rapid treatment of uncomplicated and Plasmodium falciparum malaria. The effects if oral DHA on the Lungs of albino rats were investigated.Four dosage regimens of oral DHA were tested on adult albino rats for five days and seven days. One of these doses was tested twice on one of the experimental groups. The tested dosages were 1mg/kg; 2mg/kg; 60mg/kg and 80mg/kg. The 1mg/kg dose was repeated to the test group after a one week rest period as the fifth experiment.The histological tissue photomicrographs of the study showed that oral DHA produced dose; concentration and time dependent strong stimulation of the erythropoietic stem cells in the lungs of the DHA-treated rats which was absent in the photomicrographs of control rats. The findings of this study show that oral dihydroartemisinin has a first potential usefulness in blood expansion in anaemia and a second potential usefulness in cancer treatment of cancerous blood cell diseases (possibly leukaemia), as anti-cancer effects of dihydroartemisinin have been demonstrated in many studies. Key Words: Dihydroartemisinin, Stimulatory effects, Stem cells, lungs INTRODUCTION Tremendous success of malaria treatment with dihydroartemisinin and artesunate has been recorded1, 2, 3, 4. Safety of oral treatments with dihydroartemisinin has also been noted 10. As there is still no consensus on the mechanism of action of dihydroartemisinin, studies on the effects of DHA on host systemic organs seem relevant. The present study investigated the effects of oral DHA on the lungs of Wistar albino rats. MATERIALS AND METHODS Dihydroartemisinin made by Beigin COTEC was used for the study. Five rats which weighed 104-106 grams and 75-90grams were evaluated for the effects of four oral dosage regimens and a repeated dosage regimen of dihydroartemisinin (DHA).Four rats of equivalent weights as the test rats were given orally administered distilled water to serve as controls in each experiment.
1Dept.

The tested doses of DHA were 1mg/ kg DHA; 2mg/ kg DHA; 60mg/kg DHA; and 80mg/kg DHA. A group of rats were tested twice with the 1mg/kg dosage regimen of DHA. The rats which weighed 75-90 grams received 1mg/kg rat weight of DHA for 5 or 7 days, rested for 0ne week and received the same dosage regimen again for 5days or 7 days. The rats which weighed 104-106gms received the 1mg/kg, 2mg/kg, 60mg/kg or the 80mg/kg rat weight of DHA regimen for 5 or 7 days. The rats were acclaimed for two weeks and were fed drinking water and rat chow ad libitum throughout the period of the experiments. The lungs of the test and control rats were harvested after gross anatomical observations of the organs in situ. Tissue photomicrographs of the lungs of DHA-treated and control rats were prepared using standard procedures. RESULTS Histological observation of tissue micrographs of the DHA-treated and control rats showed that DHA stimulated haemopoiesis stem cells in haemopoietic sites in the lungs shown in figures 1-3. The stimulatory action of DHA at these germ tissue sites was dose, repetition and time dependent. The stem cell stimulatory activity of DHA was identified through their initiation of cellular multiplication activity at a point that was followed by an exponential expansion outwards and upwards of the newly proliferated tissue. DISCUSSIONS Erythropoietic stem cell stimulatory activity of DHA occurred at sites indicated by arrows in figures 1-3. The stem cell stimulatory activity of DHA was signified by the initiation of cellular multiplication activity at a point and its exponential expansion outwards is observable at the sites pointed to by the arrows in fig. 1-3.

The exponential multiplicative and growth activity of the erythropoietic stem cell sites in response to the stimulatory action of DHA is well illustrated by the most visible site in fig. 3 marked by double arrows. This site in fig. 3 demonstrates that these oral DHA treatment- stimulated increases in erythrocyte and leucocyte stem cell multiplication, growth and expansion outwards (activity) were observable histologically These oral DHA stimulatory multiplication and maturation effects on the erythrocyte and leucocyte stem cells produced statistically significant (P0.01, P0.05) increases in the white blood cell count and in the packed cell volume which were similarly dose; repetition and time dependent as the histologically observable stimulatory effects on the erythropoiesis stimulation stem cells. For both types of responses, the 2mg/kg dose of DHA produced the maximal effect7. Thus, the erythrocyte and leucocyte population increases stimulated by oral DHA treatment followed the same pattern as the growth, proliferation and maturation of the erythrocyte and leucocyte germ (precursor) tissues seen in photomicrographs of the lungs of DHA-treated rats in fig. 1-3. These erythropoietic stem cell and blood cell population increases were absent in control rats. CONCLUSIONS The findings of this study show that Dihydroartemisinin directly stimulated the erythropoietic stem cells which multiplied; grew and matured to result in the reported statistical increases in the erythrocyte and leucocyte population. These findings suggest that the anti-cancer effects of dihydroartemisinin 5, 6, 7, 8, 9might be of use in the treatment of cancer that involves generation of red or white blood cells apart from the fact that these de-novo erythropoiesis- stimulatory effects of DHA on erythropoietic stem cells of the lungs are useful for the treatment of anaemia. REFERENCES AND NOTES
1. World Health Organization. (1994-1995).The role of artemisinin and its derivatives in the current treatment of malaria (WHO/MAL/94.1067), WHO, Geneva. 2. Cumming J. N., Ploypradith P. Posner G. H. (1997). Antimalarial Activity of Artemisinin (qinghaosu) and related Trioxanes: mechanism of action. Adv.J. Pharmacol, 37:253-297 [Pub Med]. 3. Asawamahaskda W., Ittratt I. Pu Y-m, Ziffer H. and Meshnick S. R. (1994).Reaction of antimalarial endoperoxides with Specific Parasite Proteins, Antimicrob. Agents Chemother. 38:1854-1858 [PubMed]. 4. MHTML. (2008).THE USE OF ANTIMALARIAL RUGS, Part II: 1.8.Artemisinin And Its Derivatives mht MHTML Document 9/19/2008.Ahsan Manan Khan Bhutta Internet Explorer. 5. Huan- Huan Chen, Hui-Jun Zhou and Xin Fang. (2003).Inhibition of human cancer cell line growth and human umbilical vein endothelial cell angiogenesis by artemisinin derivatives in vitro. Pharmacological Research 48:231-236.

of Pharmacology and Toxicology, Nnamdi Azikiwe University, P. M. B. 5025, Awka, Anambra State, Nigeria. E-mail: unedosa@yahoo.com *Corresponding author
2Dept.

of Pharmacology and Toxicology, University of Nigeria, Nsukka, Enugu State, Nigeria.

3Evangelical

Churches of West Africa Hospital (ECWA), Egbe, P. M. B.,202, Kogi State, Nigeria.
4Department

of Medical Microbiology, University of Jos Teaching Hospital, P.M. B. 2076, Jos, Plateau State, Nigeria.
5Department

of Industrial Microbiology, Federal University of Technology, Owerri, Imo State, Nigeria.

6Medical

Instructional Unit of Jos Teaching Hospital, P.M. B. 2076, Jos, Plateau State, Nigeria.

Inventi Rapid: Molecular Pharmacology Vol. 2011, Issue 4 [ISSN 0976-3856]

2011pmp099, CCC: $10 Inventi Journals (P) Ltd Published on Web 06/10/2011, www.inventi.in

RESEARCH ARTICLE

Figure(1)

Figure(2)

Figure(3)

Figure 1-3: Photomicrographs of the lungs of oral dihydroartemisinin-treated rat in which arrows point at the erythrocyte and leucocyte stem cells stimulated by dihydroartemisinin treatment. The effect of DHA treatment caused growth, proliferation and maturation of these stem cells into new red and white blood cells

Figure 4: Photomicrographs of the lungs of a control rat

6. Efferth T., Dunstan H., Sauerbrey A., Miyachi H. and Chitambar C.R. (2001). The antimalarial artesunate is also active against cancer. Int. J.Oncol. 18:767-73. 7. Woerdebag H. J., Moskal T. A., Pras N., Kishimoto S., Sudo K., KanamaruT. and Brem H. (1993).Cytotoxicty of artemisinin-related endoperoxides to Enrlich ascites tumor cells. J. Nat. Prod. 56:849-59. 8. Lai H and Singh N. P. (1995). Selective cancer cell cytotoxicity from exposure to dihydroartemisinin and holotrasferrin. Cancer Lett.91:41-6. 9. Singh N. P. and Lai H. (2001).Selective toxicity of dihydroartemisinin and holotransferrin toward human breast cancer cells. Life Science; 79:49-56. 10. A. U. Utoh-Neddosa, P. A. Akah, T. C. Okoye and C.O. Okoli .(Evaluation of the Toxic Effects of Dihydroartemisinin on the Vital Organs of Wistar Albino Rats, American Journal of Pharmacology and Toxicology 4:169-273. 11. Nosten F. C, Luxemburger and F. O. Ter Kuile. (1994).Treatment of Multidrug-resistant Plasmodium falciparum malaria with 3-day artesunate mefloquine combination. J. Infect. Disease, 170: 971-977.

Inventi Rapid: Molecular Pharmacology Vol. 2011, Issue 4 [ISSN 0976-3856]

2011pmp099, CCC: $10 Inventi Journals (P) Ltd Published on Web 06/10/2011, www.inventi.in