Effect of Potassium Dichromate on Ovary and Uterus in Female Mice

 

Abdul-Hadi AbbasHadi^1*, Kareema Habeeb Dohan²

¹Department of Laboratory Investigations, Faculty of Science, University of Kufa, Iraq

²Department of Biology, Faculty of Science, University of Kufa Iraq

 

ABSTRACT:

The histopathological alterations in ovary and uterus were evaluated to investigate the possible effects ofhexavalent chromium as potassium dichromate (K₂Cr₂O₇)in female albino mice.Forty-five adult female albino mice were used in this study.The mice treated orally with 500 and 1000 ppm of potassium dichromate that dissolved in tap water for 10, 20, and 30 days.The histologic examination by light microscope for the ovaries and uteri in potassium dichromate-treated mice revealed different histopathological alterations in these organs, whilst these organs of control groups exhibited a normal architecture. In addition, the results showed that the histological changes depend on the concentration and duration of exposure to potassium dichromate.

 

 

INTRODUCTION:

Chromium (Cr)  is the sixth most abundant element in the earth’s crust, where it is joined with iron and oxygen as chromite ore⁽¹⁾.Trivalent chromium (Cr³⁺) is the most stable oxidation state in which chromium is found in living organisms. Hexavalent chromium(Cr⁶⁺) is the second most stable form and a strong oxidizing agent, especially in acidic media. Hexavalent chromium is bound to oxygen as chromate (CrO₄^2–) or dichromate (Cr₂O₇^2–) with a strong oxidative capacity⁽²⁾.The biological systems require minute  amounts of chromium element for their normal functions. In contrast, the unsafe industrial usage for chrome can lead to pollution of the environment and to unfavorable various effects of organisms⁽³⁾. the hexavalent chromium is a toxic compound and has a potential toxicity for organisms⁽⁴⁾. The environmental pollution, occupational problems and health hazards of exposure to chromium compounds in animals and human were well documented in several reports^{(1, 2, 3, 4, 5,6,7,8)}.

 

Moreover, the developmental and reproductive toxicity of chromium are summarized by OEHHA (2009)⁽⁹⁾ and  Marouani etal. (2015)⁽¹⁰⁾.Thus, the purpose of the current study was to investigate the possible effects of potassium dichromate on histological aspects of ovary and uterus in albino mice.

 

MATERIALS AND METHODS:

Fort-five adult female albino mice (Mus musculus) were used in this study. The present study was conducted at the animal house and animal laboratories of Faculty of Science /University of Kufa.  Healthy mice weighing between 35-40 gm were used in this experiment. The animals were maintained in an air-conditioned room in separated plastic cages at controlled environment of 22-25 °C throughout the study. The animals received pellets and tap water adlibitum.

 

The female mice were randomly distributed into three groups (15 mice in each group). Each group was subdivided into three secondary groups (5 mice in each group) in separated cages. Tow concentrations of potassium dichromate (500 ppm and 1000ppm) dissolved in clean tap water were investigated. The orally concentrations as drinking water of potassium dichromate were determined according to Trivedi etal. (1989)⁽¹¹⁾. The control group received only drinking water. The chromium treated groups were examined for three progressive time periods (10, 20, and 30 days) from the beginning of exposure.

 

The reproductive effects of potassium dichromate were evaluated in estrus phase.At the end of the experimental periods (10, 20, and 30 days), all mice were anesthetized, using a mixture of ketamine and xylazine i.m , and then they were sacrificed ⁽¹²⁾. For histopathological study, ovaries and uteri were removed and cleaned. Thereafter, these organs were fixed immediately in 10% formalin solution for later histological preparation.Ordinary histological technique was followed to prepare slides from specimens of uteri and ovaries from all animal groups to study the alterations that may be found in chromium-treated animal groups. The preparation of microscopic slides and staining techniques were performed according to Bancroft and Stevens (1982)⁽¹³⁾.

 

The stained sections of ovaries and uterus from each animal were observed under the compound microscope to evaluate the histological features and structural changes. Photomicrographs were taken by camera mounted microscope at different magnifications .

 

RESULTS AND DISCUSSION:

The results of present study demonstrated that administration of potassium dichromate at concentration 500 and 1000 ppm for 10, 20 and 30 days produced a variety of histopathological changes in the ovaries and uteri of exposed animals as compared to normal histological examination of ovary  and uterus tissues in control mice.Furthermore, this study showed that the histological changes were observed in treated animals depend on the concentration and duration of exposure to potassium dichromate, and the  concentration 1000 ppm for 30 days had pronounced effects on histological features of these organs.

 

Histologic study of ovary:

The microscopic examination of ovary sections of control mice showed normal histological structure. In contrast, the gross histopathological changes in the ovaries in treated mice with potassium dichromate included retarded growth of primary follicles, degeneration and necrosis of primary and secondary follicles, arrested follicular growth and development at the secondary follicular stage, absence of antral cavity in some growing follicles,large numbers of atretic follicles throughout the stromal spaces, distorted nucleus in oocytes, presence of edematous fluid in the follicular antrum, abnormality arrangements of thecal cells and granulosa cells, pyknotic nuclei of the follicular cells, dilated blood vessels, and hemorrhage in stromal spaces. (Figures 1, 2, 3, 4, 5 and 6).

 

Our findings revealed different histological changes in ovarian tissue  of mice. These abnormalities may be related to the accumulation and direct effects of potassium dichromate in this organ. The recent studies demonstrated that chromium levels in ovary tissue for exposed rats and pigs were significantly higher than the control animals ^{(10, 14,15,16)}.

 

 

Figure (1): Ovary section of control group shows ovarian follicles (OF), corpus luteum(CL)and normal distribution of cellular elements. (H and E, 40x).

 

 

Figure (2): Ovary section of potassium dichromate-treated group (500 ppm) for 20 days showsretarded growth (RG) of primary follicles,and hemorrhage (H) in stromal spaces. (H & E, 100x).

 

 

Figure (3): Ovary section of potassium dichromate-treated group (500 ppm) for 30 days showsdegeneration (D) of oocytes, arrested follicular growth and hemorrhage (H). (H and E, 200x).

 

Figure (4): Ovary section of potassium dichromate-treated group (1000 ppm) for 10 days shows presence of edematous fluid (EF) in the follicular antrum and absence of antral cavity in some growing follicles (H and E, 40x).

 

Figure (5): Ovary section of potassium dichromate-treated group (1000 ppm) for 20 days showspyknosis (P) of follicular cells and abnormality arrangements of thecal cells and granulosa cells (H and E, 100x).

 

Figure (4-22): Ovary section of potassium dichromate-treated group (1000 ppm) for 30 days shows sever damage in ovarian tissue,large numbers of atretic follicles (AF) and dilatation (D) in blood vessels (H and E, 40x).

 

Recently, it has been reported that chromium compounds causes oxidative damage in ovarian tissues of treated animals by increasing the generation of free radicals and or by decreasing antioxidant levels in the body ^{(10, 15,17,18)}. Consequently, the free radical mechanism may be represent the cause of pathologic changes in ovary.

 

According Marouani et al. (2015)⁽¹⁰⁾, any effect of xenobiotics such chromium(VI) on the stages of follicular maturation and differentiation reflects an effect on the ovarian physiology, thereby impairing the normal reproductive competency in affected females. In previous report, it may be postulated that chromium affects the follicular maturation and differentiation in ovary of mice ultimately resulting in a reduction the number of ova released ⁽¹⁹⁾. Another study indicates that chromium(VI) causes early reproductive senescence by increased germ cell apoptosis, depleting the healthy primordial follicle pool resulting in a reduced follicular reserve and increasing follicular atresia ⁽²⁰⁾.

 

Recent reports showed that chromium(VI) decreased the ovarian granulosa cell proliferation through cell cycle arrest and decreased cyclin-dependent kinases (CDKs)⁽²¹⁾. Results of Banu etal. (2011)⁽²²⁾ indicated that chromium(VI) induced DNA fragmentation and apoptosis of granulosa cells in a time-dependent manner through cytochrome C and caspase-3 dependent intrinsic apoptotic pathways.

 

Histologic study of uterus:

The histological appearance of uterus in potassium dichromate-treated mice showed structural alterations in comparison with the control groups. The main histopathological changes observed in uteri were hyperplasia in the lamina propria of endometrium, thickening of endometrium folds, cytoplasm vacuolization and degeneration in some cells of the columnar epithelium of endometrium, degeneration in the cellular elements of connective tissue and necrosis of cuboidal epithelial cells of uterine glands, atrophy of smooth muscles resulting in decreased thickness of  myometrium, dilatation in blood vessels, destruction the walls of uterine arteries, hemorrhage and presence of edematous fluid. (Figures 7, 8, 9, 10, 11 and 12).

 

There is limited information on the effect of potassium dichromate on the uterine tissue in mice. Recently, Samuel et al. (2011)⁽²³⁾ found that chromium(VI) level in both blood serum and uterine tissue was higher in lactational  rats that treated with chromium.

 

Figure (7): Uterus section of control group shows perimetrium (P), myometrium (M), endometrium (E), and normal architecture of uterine tissue (H and E, 40x).

 

Figure (8): Uterus section of potassium dichromate-treated group (500 ppm) for 20 days showsthickening (T) of endometrium folds and fibrous tissue proliferation (P). (H and E, 40x).

 

Figure (9): Uterus section of potassium dichromate-treated group (500 ppm) for 30 days shows a damage (D) in uterine glands andpresence of edematous fluid (EF) (H and E, 100x).

 

Figure (10): Uterus section of potassium dichromate-treated group (1000 ppm) for 10 days shows necrosis (N) incolumnar epithelium of endometrium,atrophy of myometrium anddilatation (D) in blood vessels, (H and E, 200x).

 

Figure (11): Uterus section of potassium dichromate-treated group (1000 ppm) for 20 days showshyperplasia (H) in the lamina propria of endometrium and abnormal features (AF) of  uterine glands (H and E, 100x).

 

Figure (12): Uterus section of potassium dichromate-treated group (1000 ppm) for 30 days showscytoplasm vacuolization (CV) in epithelium of endometrium and degeneration (D)  in the cellular elements myometrium. (H and E, 200x).

 

Kumar etal. (2013) ⁽²⁴⁾ have reported that administration of chromium resulted in oxidative stress in female reproductive system of rats that was reflected by altered histoarchitecture, with atrophy of endometrial glands in uterus, hyperplasia of uterine epithelium and fibrous tissue proliferation. Recent reports stated that the subacute treatment with chromium(VI) causes oxidative stress in uterus inducing epithelial and stromal cells apoptosis by activation of Bax and p53 proteins. Moreover, increased reactive oxygen species (ROS) production in uterus may be responsible of the deleterious effects observed in the uterus cells ^(10,25).    

Finally, it has been proposed that the histological damage observed in uterus could be the result of increased ROS level in uterus and/or of decreased steroid hormones secretion particularly estrogen. Thus, the subacute treatment with chromium(VI) affects the ovary and uterus and causes delayed follicular development and extended estrous cycle with impaired blood levels of follicle stimulating hormone (FSH) and luteinizing hormone (LH) ⁽¹⁰⁾.

 

CONCLUSION:

Oral exposure of hexavalent chromium as potassium dichromate at 500 and 1000 ppm via drinking water for 10, 20, and 30 days in mice induced marked impact on the ovaries and uteri in various levels.It is therefore necessary to take precautions when using chromium compounds in common applications for a long time to avoid the chromium toxicity.

 

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Accepted on 20.11.2017          © RJPT All right reserved

Research J. Pharm. and Tech 2018; 11(2):429-433.