Printer Friendly

Dose-Dependent Morphological Changes of Cadmium Chloride on Kidney of Albino Mice.

Byline: Ghumman N.A., Naseem N., Latif W. and A.H. Nagi

Keywords: Cadmium, nephrotoxicity, crescentic nephropathy, interstitial inflammation.

INTRTODUCTION

There are various causes of toxicity among which heavy metals such as Cadmium (Cd) is the most important nephrotoxic substances. Toxicants manifest their toxicity in specific target organs as reported by European Commission and the World Health Organization1 with kidney being the most susceptible one. Cadmium was discovered by Friedrich Stromeyer in 1817 in Germany as a contaminant in Zinc carbonate.2 It is one of the poisonous heavy metals obtained as a byproduct of mining and refining zinc.3 Cadmium is a momentous toxin present in our environment.4 Fundamental sources of cadmium are soil,5 contaminated water and air by the procedure of mining, smelting and industrial waste.6 Cadmium is enormously being utilised at conventional industrial level.

The kinds of employees probably exposed to cadmium include manufacturersof alloys, aluminum, automobile, batteries, cadmium plates, ceramic and pottery, Cd-copper alloys, dental amalgam, electroplates, electrical condensers, electric instruments, glass, jewelry, luminous lamps, lithophane, pesticides, paints, photoelectric cells, pigments, plastics and many others.7 Cadmium is taken-up from the lungs and gastrointestinal tract and is transported via blood to liver and kidneys. Within the liver, production of metallothionein is initiated by the cadmium. Cadmium that is bound to metallothionein once released into plasma will be filtered into renal tubular fluid. Metallothionein is the protein that stores cadmium in body and plays a central role in the transportation of cadmium from liver via blood to kidneys.8

This metal accumulates within the kidneys and liver and incorporates a long biological half life of seventeen to thirty years in human being.9 After long-term cadmium exposure, renal tubular dysfunction may develop both in experimental animals and human.10 Cadmium exerts its toxic effects on the kidneys, respiratory system and the skeletal system and is classified as a human carcinogenby the International Agency for Research on Cancer (IARC), which is part of the World Health Organization.4 In occupationally exposed population, preliminary signs of glomerular devastation from cadmium are escalated elimination of high mass proteins like iron binding glycoproteins and albumin. Degree of detrimental effects on glomeruli is dose-dependent and once started, the glomerular damage is believed to be irreversible.11 In Japan, post-menopausal women who were exposed to immoderate concentrations of cadmium over their lifespan, "Itai-itai" or ouch-ouch disease was observed.

In these females, the primary source of cadmium exposure was through their diet, because those residential areas of Japan were significantly polluted with cadmium12. Signs and symptoms of "itai-itai" disease comprise of osteomalacia and alarming osteoporosis with concurrent grievous renal dysfunction.Cd levels in urine, blood, faeces, hair, kidney, liver and different tissues have been utilised as biological indicators of cadmium exposure.13 Based on an extensive exposure to this toxic metal this study was designed to observe the morphological changes in kidneys of Albino Mice induced by different doses of oral preparations of anhydrous cadmium chloride.

MATERIALS AND METHODS

It was an experimental interventional, randomized controlled study in adult mice. Forty Eight male and female albino mice of BALB/c strain, 6 - 8 weeks old weighing 30 +- 5g, were included in the study. Animals were separated gender wise in different cages and maintained in the Animal House of the University of Health Sciences, Lahore under controlled environment (temperature 22-25AdegC, humidity 65% +- 5) and light and dark cycle of 12 hours each. Albino mice were segregated in 4 groups with one control group and 3 experimental groups each comprising of 12 mice. In this foregoing experiment, cadmium was used as cadmium chloride (CdCl2) orally on alternate days for 8 weeks. According to the body weight (5 mg/kg body weight) the dose was calculated and mixed with distilled water14 (Table 1). The control group was given normal diet and plain tap water. Serum creatinine was measured at the end of the experiment by using commercially available kits (RandoxCR510, LOT: 216982).

Urinary proteins were determined by strip method (Roche Diagnostic GmbH). Blood samples from each group were collected by cardiac puncture. At the end of the experiment, mice were sacrificed and kidneys were removed immediately, fixed in 10% formalin, routinely processsed and stained with haematoxylin and eosin, Periodic Acid Schif staining and Periodic Acid Schiff-Methenamine Silver stain.

Statistical Analysis

The data was entered and analysed using SPSS 21.0. Mean values were taken for quantitative variables (Serum creatinine and urinary proteins). Frequencies and percentages were given for qualitative variables (histopathological changes in kidney). Fisher's exact test was applied.

RESULTS:

A total of 48 male and female albino mice of 6-8 weeks were taken. The animals were distributed into four groups with 12 mice in each group as A, B, C and control group. After a week of acclimatisation, the experiment was started. After a period of 8 weeks, the experiment was terminated and animals were sacrificed after taking blood sample via cardiac puncture. Results were analysed using Fischer's Exact test and P value was found to be significant in case of all the variables (Table 2, 3, 4).The changes noticed were mainly shown in the tables below. These include biochemical changes like proteinuria and S/Creatinine (Fig. 1-2). Histopathological changes like glomerular adhesions, glomerular crescent formation, tubular necrosis, tubular vacuolar degeneration, mesangial hypercellularity, interstitial fibrosis, vascular sclerosis, capillary wall thickness, tubular cast and interstitial inflammation.

Many other histopathological changes were also noticed in the animals of group C such as glomerular sclerosis, mesangial widening, vascular congestion, perivascular inflammation, swelling of the nuclei of the tubular epithelaial cells, fibrinoid change in the afferent arterioles, lobulation of glomeruli and atrophy of glomeruli, however these changes were not observed in a significant number of animals that could be elaborated (Fig. 3-8).

Table 1: Groups of experimental animals.

###Group###Mice###Intervention###Dosage/ Alternate Day###Route###Duration

###Control###12###Normal diet###None###Oral###8 weeks

###A###12###CdCl2###5 mg/kg###Oral###8 weeks

###B###12###CdCl2###10 mg/kg###Oral###8 weeks

###C###12###CdCl2###15 mg/kg###Oral###8 weeks

Table 2:

Histological Variables

Glomerular Adhesions

###Absent###50%

Groups

###n(%)###n(%)###n(%)###n(%)

Control###12 (100)###0 (0.0)###0 (0.0)###0 (0.0)

###A###2 (16.7)###6 (50.0)###4 (33.3)###0 (0.0)

###B###0 (0.0)###6 (50.0)###6 (50.0)###0 (0.0)

###C###0 (0.0)###5 (41.7)###5 (41.7)###2 (16.7)

Fisher's Exact Test = 39.689; p-value < 0.001

Glomerular Crescent Formation

Control###12 (100)###0 (0.0)###0 (0.0)###0 (0.0)

###A###3 (25.0)###7 (58.3)###2 (16.7)###0 (0.0)

###B###3 (25.0)###3 (25.0)###6 (50.0)###0 (0.0)

###C###7 (58.3)###2 (16.7)###3 (25.0)###0 (0.0)

Fisher's Exact Test = 22.176; p-value < 0.001

Tubular Necrosis

Control###12 (100)###0 (0.0)###0 (0.0)###0 (0.0)

###A###0 (0.0)###8 (66.7)###3 (25.0)###1 (8.3)

###B###0 (0.0)###4 (33.3)###7 (58.3)###1 (8.3)

###C###0 (0.0)###0 (0.0)###8 (66.7)###4 (33.3)

Fisher's Exact Test= 54.043;p-value < 0.001

Tubular Vacuolar Degeneration

Control###12 (100)###0 (0.0)###0 (0.0)###0 (0.0)

###A###0 (0.0)###10 (83.3)###2 (16.7)###0 (0.0)

###B###0 (0.0)###6 (50.0)###5 (41.7)###1 (8.3)

###C###0 (0.0)###7 (58.3)###5 (41.7)###0 (0.0)

Fisher's Exact Test= 46.609;p-value < 0.000

Table 3:

Mesangial Cellularity

###Normal###Mild###Moderate###Severe

Groups

###n(%)###n(%)###n(%)###n(%)

Control###12 (100)###0 (0.0)###0 (0.0)###0 (0.0)

###A###0 (0.0)###6 (50.0)###6 (50.0)###0 (0.0)

###B###0 (0.0)###0 (0.0)###7 (58.3)###5 (41.7)

###C###0 (0.0)###0 (0.0)###4 (33.3)###8 (66.7)

Fisher's Exact Test= 59.369;###p-value < 0.001

Vessel Thickness (Sclerosis)

Control###12 (100)###0 (0.0)###0 (0.0)###0 (0.0)

###A###12 (100)###0 (0.0)###0 (0.0)###0 (0.0)

###B###12 (100)###0 (0.0)###0 (0.0)###0 (0.0)

###C###6 (50.0)###6 (50.0)###0 (0.0)###0 (0.0)

Fisher's Exact Test= 32.619;p-value < 0.005

Table 4: Glomerular Capillary Wall Thickness after 8 Weeks.

###Normal###Focally Thick###Diffusely Thick

Groups

###n (%)###n (%)###n (%)

Control###12 (100)###0 (0.0)###0 (0.0)

###A###4 (33.3)###8 (66.7)###0 (0.0)

###B###2 (16.7)###3 (25.0)###7 (58.3)

###C###0 (0.0)###3 (25.0)###9 (75.0)

Fisher's Exact Test= 42.45; p-value 50% necrosis in mice of group C, B and A, as well as a few of them showed tubular necrosis upto 25-50% (Figure 7)and these consequence of cadmium are well established.21

Regarding tubular vacuolar degeneration, it was also found in all the experimental mice. This damage was in accordance with the other results mentioned above with more damage in group C followed by B and A (Figure 3) as described by other researchers.22,18,20 In mice, cadmium also induced deposition of eosinophilic casts, mainly protein cast in the tubules (Figure 4). Severe interstitial inflammation was also a commendable observation in this study. The degree of interstitial inflammation was exactly in accordance with the other results of this study revealing that it was dose dependent in all groups. So majority of the group C animals were found to develop moderate to severe inflammation (Figure 3) as narrated in the past studies by Prozialeck et al. in 2009.19

Blood vessels were also not spared and developed thickness of their wall (sclerosis). Vascular sclerosis by the cadmium toxicity have been explored by Messner and Bernhard (2010)23 that showed that cadmium is a distinctive risk factor for vascular disease.

It is concluded that Cadmium is a potential nephrotoxic chemical and can lead to progressive renal failure by inducing dose-dependent pathological changes in glomeruli. In this study pathological changes in proximal tubules, interstitium and blood vessels are also observed culiminating to tubulointerstitial fibrosis. As this chemical is a naturally occurring toxicant present in air, water, soils, and foodstuffs, hence its emission may be reduced and monitored for better healthcare of community and environment.

ACKNOWLEDGMENT

I would like to express my unrestrained appreciation to the Vice chancellor of University of Health Sciences Lahore for supporting this research.Sincere thanks also go to Mr. Sameer Anjum and Mr. Usman Ali and Mrs. Sadia Hasnat who helped me and guided me regarding my laboratory work.

Author's Contribution

NAG: Substantial contribution of concept and design of work, write-up, analysis and data interpretation and results compiling. NN: Supervision and revising it critically for important intellectual content. WL: Help in biostatics and data analysis. AHN: Overall supervision in concept and design of work and final approval of the work.

Funding Source

University of Health Sciences, Lahore.

Conflict of Interest: There is no conflict of interest.

REFERNECES

1. European Commission and the World Health Organization International Programme on Chemical Safety. Proceedings of the International Workshop on the health significance and early detection of nephrotoxicity. Toxicol. Let. 1989; 46: 1-301.

2. Morrow H. "Cadmium and Cadmium Alloys". Kirk-Othmer Encyclopedia of Chemical Technology, 2010: pp. 1-36.

3. The Element Cadmium. Cadmium-element properties and periodic table information 2012. "[online] Available at: . [Accessed 13th July 2018].

4. Satarug S, Garrett SH, Sens MA, Sens DA. Cadmium, environmental exposure, and health outcomes. Environ. Health. Perspect. 2010; 118: 182-190.

5. Agency for Toxic Substances and Disease Registry (ATSDR). Toxicological Profile for Cadmium. US Department of Human and Health Services, 1999.

6. Casado M, Anawar HM, Garcia-Sanchez, A, Santa-Regina I. Cadmium and zinc in polluted mining soils and uptake by plants (El Losar mine, Spain). Int. J. Environ. Pollut. 2008; 33: 146-159.

7. National Institute of Occupational Safety and Health. Testimony on Occupational Exposure to Cadmium, 1990.

8. Zalups RK, "Molecular handling of cadmium in transporting epithelia". Toxicol Appl Pharmacol. 2003; 186 (3): 163-188.

9. Nordberg G, Nogawa K, Nordberg M, Friberg L. Cadmium.In: Handbook on toxicology of metals. Academic Press, 2007: 65-78.

10. Barbier O, Jacquillet G, Tauc M, Cougnon M, Poujeol P: Effect of heavy metals on, and handling by, the kidney. Nephron Physiol. 2005; 99 (4): 105-10.

11. Jarup L. "Cadmium overload and toxicity." Nephrology Dialysis Transplantation, 2002; 17 (Suppl. 2): 35-39.

12. Ikeda M, Ezaki T, Moriguchi J, Fukui Y, Ukai H, Okamoto S, Sakurai H. "The threshold cadmium level that causes a substantial increase in urine of general populations." Tohoku. J. Exp. Med. 2005; 205: 247-261.

13. Lauwerys RR, Bernard AM, Roels HA, Buchet JP. Cadmium: Exposure markers as predictors of nephrotoxic effects. Clin Chem. 1994; 40 (7): 1391-1394.

14. Prabu MS, Selvarajan N, Hemalatha S, Rameshkumar T. Hepatoprotective effect of Andrographiespaniculata against cadmium induced toxicity in male Wistar rats. Toxicol. Int. 2008; 15: 21-25.

15. Waisberg M, Joseph P, Hale B, Beyersmann D. Molecular and cellular mechanisms of cadmium carcinogenesis. Toxicology, 2003; 192: 95-117.

16. Sanchez-Chardi A, Penarroja-Matutano C, Borras M, Nadal J. Bioaccumulation of metals and effects of a landfill in small mammals Part III: structural alterations. Environ. Res. 2009; 109: 960-967.

17. Thomas LD, Hodgson S, Nieuwenhuijsen M, Jarup L. Early kidney damage in a population exposed to cadmium and other heavy metals. Environ. Health. Perspect. 2009; 117: 181-184.

18. Abdel-Moneim AM, Said KM. Acute effect of cadmium treatment on the kidney of rats: biochemical and ultra-structural studies. Pak. J. Biol. Sci. 2007; 10: 3497-3506.

19. Prozialeck WC, Edwards JR, Lamar PC, Liu J, Vaidya VS, Bonventre JV. Expression of kidney injury molecule-1 (Kim-1) in relation to necrosis and apoptosis during the early stages of Cd-induced proximal tubule injury. Toxicol. Appl. Pharmacol. 2009; 238: 306-314.

20. Kukner A, Colakoglu N, Kara H, Oner H, Ozogul C, Ozan E. Ultrastructural changes in the kidney of rats with acute exposure to cadmium and effects of exogenous metallothionein. Biol. Trace Elem. Res. 2007; 119: 137-146.

21. Erfurt C, Roussa E, Thevenod F.. Apoptosis by Cd2+ or CdMT in proximal tubule cells: different uptake routes and permissive role of endo/lysosomalCdMT uptake. Am. J. Physiol. Cell Physiol. 2003; 285: C1367-C1376.

22. Jihen H, Imed M, Fatima H, Abdelhamid K. Protective effects of selenium (Se) and zinc (Zn) on cadmium (Cd) toxicity in the liver and kidney of the rat: histology and Cd accumulation. Food Chem. Toxicol. 2008; 46: 3522-3527.

23. Messner B, Bernhard D. Cadmium and cardiovascular diseases: cell biology, pathophysiology, and epidemiological relevance. Biometals. 2010; 23: 811-822.
COPYRIGHT 2018 Knowledge Bylanes
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2018 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Publication:Biomedica
Date:Dec 31, 2018
Words:2718
Previous Article:Comparative Study of Betulinic Acid Versus Simvastatin on Total Cholesterol and HDL in Hyperlipidemic Model.
Next Article:Fatphobia: An Emerging Obsession among Students of Private Medical College, Lahore, Pakistan.
Topics:

Terms of use | Privacy policy | Copyright © 2022 Farlex, Inc. | Feedback | For webmasters |