INTRODUCTION:

Entamoeba histolytica is a gastrointestinal protozoon caused amoebiasis disease for persons by contaminated food and water with cyst or trophozoite. The introduction of the parasite does not necessarily develop into symptoms, a bigger percentage of infected persons are asymptomatic and they can be in this state for as long as a year but after signs seem, the disease can be dangerous because it may present diarrhea which may lead to acute dehydration^1,2.

Phagocytosis starts with the ligation of the Galactose/ N-Acetyl galactosamine (Gal/GalNAc lectin) lectin³ and in order to promote degradation, amoebapores and cysteine proteinases are secreted to the phagosome².

There are evidences of the main role of cysteine proteinases as a virulence factor for E. histolytica being involved in the breach of the mucus barrier, which is crucial in the pathogenesis of amoebiasis⁴.

E. histolytica is needs a high level of iron to continue, this protozoan pathogen is capable to get iron from host proteins, example hemoglobin, ferritin, Lactoferrin and transferrin. That E. histolytica trophozoites endocytose ferritin by clathrin-coated pits and degrade this protein by means of specific cysteine proteases in the endosome/lysosome pathway⁵. Ferritin is a hetero polymer composed of 24 subunits of two types, H and L, and the proportion of each subunit depends on the main function of the protein. For example, in the liver and spleen⁶. The molecular mechanisms of iron acquisition in protozoa are still poorly understood, iron storage is extremely important to all forms of life, thus ferritin is found in the three domains of living beings, Bacteria, Archaea, and Eukarya. However, some eukaryotic cells like yeasts and protozoa apparently do not have a ferritin molecule, but have evolved other ways to store and maintain their own iron homeostasis^{7, 8, 6}.

Many parasitic pathogens, including Trypanosoma, Leishmania and Entamoeba have advanced convergent techniques of (heme-iron) gaining from this molecule of host. Heme-iron is released by digested a protein portion of hemoglobin after capturing it by pathogen protozoa through the specific surface receptors or phagocytosis^{9, 10,11}.

The aim of this work was to study the serum melatonin, leptin,iron and Lactoferrin levels in patients with amoebiasis.

MATERIAL AND METHODS:

Specimens:

60 samples were collected from patients and 30 healthy male who attended the laboratory Hospitals in AL-Najaf province, Stool samples were collected from male patients and serum preparing from blood samples were also collected from the same patients by vein-puncture into sampling tubesby centrifugation at 3000 rpm for 5 minutes (Backman/counter, Germany) and divided the serum into four parts; each of them was kept in deep freeze at -20ºC till used for the determination of melatonin, leptin, iron and Lactoferrin. All samples collection was accepted by the Institutional Ethics Committee of the Faculty of Science at the University of Kufa and all participants signed informed consent forms.

Specimen processing:

Samples of stool were examined by light microscope using X40 objectivelens for diagnostic the intestinal parasites according to¹².

The Kits:

The serum melatonin concentration, serum leptin and serum Lactoferrin in the current Study were estimated by Eliza Kits example melatonin (Immuno – Biological Laboratories, IBL-America. RE52131), leptin (Ray Biotech, Inc. China with number code Cat#: ELH-Leptin-001), Lactoferrin (Kono Biotech/ Bulgaria /catalogue number KN0888Hu) and spectro Kits such as Iron/ Biolabo/ France (02160Maizy France).

Statistical analysis:

Data were analyzed using the software packages Graph pad prism for Windows (5.04, Graph pad software Inc. USA), Data are presented as the mean ± standard error (SE). The comparison between the patients and control groups were analyzed by student t- test. A p-value < 0.05 was considered significant.

RESULTS:

Serum melatonin:

The results revealed significant increase (P < 0.05) in serum concentration of melatonin (26.32±1.431 pg/mL) in patients with amoebiasis as compared to control group (8.41 ± 0.302 pg/mL), as seen in (fig. 1)

Fig.1: Comparison between melatonin level of amoebiasis patients and Healthy group.

*Significant difference (P<0.05) between control group and patients

Serum leptin:

The results revealed significant decrease (P < 0.05) in serum concentration of leptin (4.867±0.530 ng/mL) in patients with amoebiasis as compared to control group (8.042 ± 1.161 ng/mL), as seen in (fig 2)

Fig.2: Comparison between leptin level of amoebiasis patients and Healthy group.

*Significant difference (P<0.05) between control group and patients

Serum iron:

The results revealed significant decrease (P < 0.05) in serum concentration of iron (43.53±0.921 µg/dL) in patients with amoebiasis as compared to control group (119.81 ± 0.613 µg/dL), as seen in (fig. 3)

Fig. 3: Comparison between iron level of amoebiasis patients and Healthy group.

*Significant difference (P<0.05) between control group and patients

Serum Lactoferrin:

The results revealed significant decrease (P < 0.05) in serum concentration of Lactoferrin (13.96±1.072 ng/mL) in patients with amoebiasis as compared to control group (22.05 ± 1.210 ng/mL) as seen in (fig. 4)

Fig.4: Comparison between Lactoferrin level of amoebiasis patients and Healthy group.

*Significant difference (P<0.05) between control group and patients

DISCUSSION:

The current study has revealed that the serum leptin, Lactoferrin and iron were significant decrease in E. histolytica infection patients compared to healthy group. While serum melatonin was significant increase in E. histolytica infection patients compared to healthy group. The decrease in iron level in patients with E. histolytica may be due to the pathogenicity of this parasite dependent on the relationship between iron concentration and adhesion of parasite on epithelial cell ¹³.Whereas the source of iron maybe from the hemolysis of red blood cells from lesion occur by the parasite. The consuming of iron by E. histolytica may cause a decrease in the iron levels¹⁴.

The results of this study showed low concentration of serum leptin in patients infected with E. histolytica parasite the reason of this lowerin serum leptin may be due to a role of leptin in host resistance against disease or infection in adult and children with congenital deficiency of the leptin receptor ¹⁵,or may be to increased susceptibility to amoebiasis due to removal of the leptin receptor in the intestinal epithelium by infection 16, ^{17, 18, 5}.Many intestinal mucosal damages occur in cases of human intestinal parasitic infections such as pathological changes in the villi of epithelial cells, inflammation, intestinal abscesses, epithelial cell destruction and ulceration in intestinal epithelial layer ¹⁹.Leukophagocytosis is one of phagocytic capacity of amoebaeare continually in connection with leukocytes in vivo and essential to damage them to continue²⁰.

The current study also revealed high serum concentration of melatonin in patient with E. histolytica in compared with control group; this increased may be due to leukocytosis occur by infection with this parasite and interaction with leucocyte which causes to increased melatonin concentration and this case well-known immunomodulator^21,22.

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Received on 14.06.2017 Modified on 14.07.2017

Research J. Pharm. and Tech 2017; 10(10):3471-3473.

DOI: 10.5958/0974-360X.2017.00620.5