Download the morphometric characteristics of the peyers` patcheslymphatic

Elena N. Morozova* et al. International Journal Of Pharmacy & Technology
Available Online through
www.ijptonline.com
ISSN: 0975-766X
CODEN: IJPTFI
Research Article
THE MORPHOMETRIC CHARACTERISTICS OF THE PEYERS' PATCHES
LYMPHATIC NODULES OF THE SMALL INTESTINE IN RATS AFTER
ADMINISTRATION OF CYCLOPHOSPHAMIDE
Elena N. Morozova*, Vitaliy N. Morozov, Svetlana V. Zabolotnaya, Aleksei V. Tverskoi
Belgorod State University 308015, Russia, Belgorod, Pobeda St. 85
Email: [email protected]
Received on 04-06-2016
Accepted on 27-06-2016
Abstract
The morphometric features of the Peyers' patches lymphatic nodules of the small intestine in rats after
cyclophosphamide injection were studied. In the experimental group the length and width of the lymphatic nodules
and germinal centers were smaller than those in the control group on the 7 and the 30 day with the most evident
changes on the 30 day of the experiment. The morphometric characteristics in the experimental group as well as in
the control one were identical by the 90 day. Basing on that we can assume these changes show adaptive abilities of
the small intestine immune system after acute immunosuppression.
Key words: rat, small intestine, Peyer's patches, lymphatic nodules, cyclophosphamide, morphometric features.
Introduction
Today the problem of immunodeficiency states has become extremely important [1, 2]. The excessive influence of
the factors which depress or stimulate the immune system causes the development of the immune deficiency. The
combined influence of toxic substances characterizes most of cases in general. The such substances can accumulate in
products both of phytogenic and animal origin [3]. Considering the fact that the intestine directly communicates with
a great number of various nutrients, coming from environment and serves as an entrance for many pathogens, mucous
associate lymphoid tissue (MALT) plays a very important part in general and local immune protection [4-7].
Cyclophosphamide, commonly used as an anti-neoplastic drug, can cause adverse side-effects including
immunotoxicity, urotoxicity and immunosuppession [8]. Immunosuppression is a state of temporary or permanent
immunity dysfunction and can make organism more sensitive to pathogens due to the damage of immune system [9].
Thus, the aim of the research was to study the morphometric peculiarities of the Peyers' patches lymphatic nodules of
small intestine in rats after cyclophosphamide injection.
IJPT| June-2016 | Vol. 8 | Issue No.2 | 14097-14101
Page 14097
Elena N. Morozova* et al. International Journal Of Pharmacy & Technology
Methods
The study was carried out on 36 white mature male rats. Animals were divided into 2 groups. The rats of Group 1
were injected cyclophosphamide intramuscularly at a dose of 200 mg/kg body weight. The rats of Group 2 were out
of toxic influence. The rats were taken out of the experiment on 7, 30 and 90 day after cyclophosphamide injection.
The formalin-fixed, paraffin-embedded blocks obtained from all rats were performed for routine histopathological
examination. The specimens of small intestine (5-6 microns) were stained with hematoxylin-eosin and examined by
automated morphometric complex comprising a light microscope Olympus CX41, a digital camera and a personal
computer with a set of application programs. Using a licensed program "Morpholog" the height and width of the
lymphatic nodules and germinal centers were measured [10]. The resulting digital data were processed by using the
program “STATISTICA”.
Main part
In the intact Group the patches surface was smooth and facing into the intestinal lumen. The adjacent areas were
covered with villi, which partially closed its surface. One row of the lymphatic nodules was found in the lamina
propria and submucosa. Germinal centers were predominantly located at the base of lymphatic nodules (Fig. 1a, b).
1
1
2
3
3
4
а
4
2
b
Figure-1. The lymphatic nodules of the Peyers' patches in the small intestine (a - on the 30 day after the
cyclophosphamide injection, b – intact rats): 1 - the dome of the lymphatic nodule, 2 - germinal center of lymphatic
nodule, 3 - peripheral zone, 4 - internodular zone. Hematoxylin & Eosin. ×400.
The height and width of the Peyers' patches lymphatic nodules of the small intestine in Group 1 were smaller than
those in Group 2 by 7.7% and 19.0% and the height and width of the germinal centers - by 21.6% and 15.0%
respectively on the 7 day of the experiment (Table 1). The same tendency was observed on the 30 day of the
experiment: the height and width of the lymphatic nodules decreased by 23.5% and 0.5%, the height and width of the
IJPT| June-2016 | Vol. 8 | Issue No.2 | 14097-14101
Page 14098
Elena N. Morozova* et al. International Journal Of Pharmacy & Technology
germinal centers - by 30.5% and 1.8% respectively. The morphometric parameters of the lymphatic nodules of Group
1 became equal to these in Group 2 on the 90 day of the experiment.
Table 1: The morphometric parameters of the Peyers' patches lymphatic nodules of the small intestine in the
mature rats of Groups 1 and 2. M±m (n=36).
The morphometric parameters of the Peyers' patches lymphatic nodules (in microns)
Parameters
7 day
Group 1
Height of the lymphatic 903.0+14.1*
nodule
Width of the lymphatic 888.0+28.9*
nodule
Group 2
978.0+31.5
30 day
Group 1
808.0+31.5*
Group 2
998.0+38.3
90 day
Group 1
786.0+14.9
Group 2
755.0+33.6
1096.0+41.6
748.0+13.0
752.0+30.0
783.0+17.6*
970.0+48.2
488.0+14.1
499.0+9.4*
390.0+14.3
513.0+25.3
625.0+15.9*
846.0+38.1
Height of the germinal 518.0+21.4*
661.0+33.0
374.0+14.0*
center
Width of the germinal 627.0+9.6*
737.0+22.7
504.0+19.4
center
Note: * - significant differences from the control data if p<0.05.
Minimal measure of the morphometric parameters of the internodular zones in Group 1 in comparison with Group 2
were detected on the 30 day of the experiment. These changes prove that cyclophosphamide has a great influence on
the T-dependent zone (internodular zone) and B-dependent zone (lymphatic nodules with germinal center) on the 7
and the 30 days of the experiment. Throughout the experimental period, the lymphatic nodules in rats of Group 1
preserved the main morphological and morphometrical features. However, the boundary between the germinal center
and the peripheral zone of the nodules became indistinct on the 30 day. This phenomenon can be explained by cell
quantity decreasing in the peripheral zone. Frequently enough there were identified Peyer’s patches lymphatic
nodules of the small intestine with double domes (Fig. 2). These changes may be connected with the too large doses
of cyclophosphamide.
3
1
2
Fig. 2. The dome of the lymphatic nodules in rats of Group 1 on the 30 day of the experiment: 1 - epithelium, 2 lamina propria of the mucosa, 3 - lymphocytes. Hematoxylin & Eosin. ×600.
IJPT| June-2016 | Vol. 8 | Issue No.2 | 14097-14101
Page 14099
Elena N. Morozova* et al. International Journal Of Pharmacy & Technology
According to the data of S.Y.C. Rolph et al. , T. Nakahara et al. the use of high-dose cyclophosphamide in clinical
practice is based on the cytostatic effects of the drug, leading to inhibition of actively proliferating cells [11, 12].
Lymphocytes are the first to undergo the cyclophosphamide influence because they refer to the most proliferative
type of cells [13]. It can explain mentioned above lesions of the Peyer's patches of the small intestine.
Conclusions
1. A high degree reactivity manifested in evident changes of morphometric characteristics of the Peyer’s patches
lymphatic nodules of the small intestine was observed after cyclophosphamide injection (200 mg/kg).
2. The most significant changes of the morphometric parameters were detected on the 7 and 30 days of the
experiment.
3. The usual morphometric parameters found at the latest stages of the experiment characterize the immune system
ability for adapting to the acute immunosuppression.
References
1.
Dimitriev, D.A. i Rumjanceva, E.G., 2002. Sovremennye metody izuchenija vlijanija zagrjaznenija
okruzhajushhej sredy na immunnuju sistemu. Gigiena i sanitarija, 3: 68-71 (in Russian).
2.
Bethany, W., Humble, M.C. and Lawrence, B.P., 2011. Environmental toxicants and the developing immune
system: a missing link in the global battle against infectious disease. Reproductive Toxicology, 31(3): 327-336.
3.
Magdzіk, V.V., 2006. Іmunіtet populjacіі: vchora, s'ogodnі, zavtra. Іnfekcіjnі hvorobi, 3: 5-13 (in Ukrainian).
4.
Ruddle, N.H. and Akirav, E.M., 2009. Secondary lymphoid organs: responding to genetic and environmental
cues in ontogeny. Journal of immunology, 183: 2205-2212.
5.
Heel, K.A., McCauley, R.D. and Papadimitriou, J.M., 2008. Peyer’s patches. Journal of Gastroenterology and
Hepatology, 12(2): 122-136.
6.
Forchielli, M.L. and Walker, W.A., 2005. The role of gut-associated lymphoid tissues and mucosal defense.
British Journal of Nutrition, 93: 41-48.
7.
Kashhenko, S.A. i Shtepa, S.Ju., 2007. Morfometricheskie pokazateli beloj pul'py selezenki polovozrelyh krys
posle vvedenija im ciklofosfana. Ukrainskij morfologicheskij al'manah, 5(4): 127-128 (in Russian).
8.
Sakthivel, K.M. and Guruvayoorappan, C., 2015. Acacia ferruginea inhibits cyclophosphamide-induced
immunosuppression and urotoxicity by modulating cytokines in mice. Journal of Immunotoxicology, 12(2): 154-163.
IJPT| June-2016 | Vol. 8 | Issue No.2 | 14097-14101
Page 14100
9.
Elena N. Morozova* et al. International Journal Of Pharmacy & Technology
Gong, Y., Wu, J. and Li, S.-T., 2015. Immuno-enhancement effects of Lycium ruthenicum Murr. polysaccharide on
cyclophosphamide-induced immunosuppression in mice. International Journal of Experimental and Clinical
Medicine, 8(11): 20631-20637.
10. Ovcharenko, V.V. i Mavrich, V.V., 2004. Komp’juterna programa dlja morfometrichnih doslіdzhen'
“Morpholog”. Svіdotstvo pro reєstracіju avtors'kogo prava na tvіr # 9604, data reєstracії 19.03.2004 (in
Ukrainian).
11. Rolph, S.Y.C., Cooley, M.S. and Sewell, W.A., 2006. Cyclophosphamide augments inflammation by reducing
immunosupression in a mouse model of allergic airway disease. Journal of Allergy and Clinical Immunology,
117(3): 635-641.
12. Nakahara, T., Uchi, H. and Lesokhin, A.M., 2010. Cyclophosphamide enhances immunity by modulating the
balance of dendritic cell subsets in lymphoid organs. Blood, 115(22): 4384-4392.
13. Gladstone, D.E., Zamkoff, K.W. and Krupp, L., 2006. High-dose cyclophosphamide for moderate to severe
refractory multiple sclerosis. Archives of neurology, 63(10): 1388-1393.
Corresponding Author:
Elena N. Morozova*,
Email: [email protected]
IJPT| June-2016 | Vol. 8 | Issue No.2 | 14097-14101
Page 14101