INTRODUCTION:

Cadmium is one of heavy metal that was found in 1817 also in the 20^th century and declared as toxic heavy metal.Cadmium can be found deep in the crust, soil from a volcano eruption, rock eruption, mining, fossil remains, plant, industry and commonly found as a mineral that binds into O₂, chlorine, and Sulfur^1,2. Cadmium intoxication that is very popular in the world known as itai-itai disease in Japan³.

The organ that keeps more Cadmium in body are liver and kidney^2,4. Liver will become damaged from acute or chronic exposure toCadmium⁵.

Some researchers show that Cadmium caused structural changing in liver macroscopically or microscopically, research done by Refaiy and Eissa in 2013, rat liver that intoxicated with Cadmium, showed fatty liver, focal necrosis, karyopyknotic, karyolysis and kupffer cell proliferation^2,
6.

Cadmium can enter the body through food, drinking water also inhalant then carried by erythrocytes and albumin. Research on experimental animal stated that Cadmium has an adverse effect on the liver, kidney, lungs, bone, reproductive organ, hematopoietic system, nerve system and cardiovascular system⁵.

Generally, the mechanism of Cadmium toxicity is caused by oxidative stress. Oxidative stress is defined as an imbalance between the amount of ROS that produced with eliminated ROS from the antioxidant system⁷.

Antioxidant and chelating agent were generally used as therapy in heavy metal toxicity. Natural antioxidant may able to give some protection to oxidative stress. Antioxidant is a broad spectrum component which able to donor electron and neutralized free radical also prevent cell damage^8,9. The search of effective, non-toxic, and derived antioxidants from natural ingredients has been intensified in these recent years¹⁰.

Curcumin is a yellow-colored pigment that can be found in a plant with genus Curcuma with percentage 76% in the extract. Curcumin is included in polyphenol compound with antioxidant characteristic also can be found in green tea¹¹. Research shows that Curcumin can be used as antiinflammation and antioxidant also considered ashepatoprotective and antifibrosis component. Curcumin stated as phytochemistry that safe to use, non-toxic, genotoxic or teratogen even used in high dosage^{12, 13}.

Curcumin is widely used because it can improve health, but only few information about the effect of Curcumin as therapy in metal intoxication include the effect on liver damage⁷.

MATERIAL AND METHODS:

Experimental animal:

The animal used in this research were 20 female mice strain aged 16 weeks old, weighed 30 gram, divided into four groups and each group consisted of 10 female mice. Male mice used was20, aged 16 weeks, and weighed 30 gram. The mice have never been ever used to research and obtained from Pusat Veterinaria Farma Surabaya.

Research Location and Time:

The research was conducted in Laboratory of Experimental Animal, Faculty of Veterinary Medicine, University of Airlangga Surabaya during two months. The histological slide was done in Department of Veterinary Pathology, Faculty of Veterinary Medicine, University of Airlangga, and the examination was conducted in Department of Veterinary Anatomy, Division of Veterinary Histology, Faculty of Veterinary Medicine, University of Airlangga.

Research equipments:

The equipment used in this research were animal cages, syringe, needle 22G, surgical instruments, petri disk, feeding tube, balancer and Nikon Eclipse Cimicroscope.

Research materials:

The material used in this research were Cadmium Chloride (Sigma Aldrich) with dosage 4.45 mg/KgBW, Curcumin (Merck), CMCNa, aquadest, physiologic saline and formalin buffer.

Research procedure:

Estrus synchronized was done on female mice by injecting PMSG 5 IU and HCG 5 IU. The female mice then mated by monomating procedure and after 17 hours then continued with vaginal plug examination. Female mice with vaginal plug mean that it is the first day of pregnancy. Pregnant mice were divided into 4 groups consisting five mice per group, with total 20 mice. T0 as a negative control group, T1 as the positive control group which injected by Cadmium 4, 5mg/kgBWi.p in gestation day 7, T2 as the group which injected by Cadmium 4, 5mg/kgBWi.pin gestation day 7 then Curcumin therapy 150mg/kgBW were administered p.o ten days consecutively and T3 as the group which injected by Cadmium 4, 5mg/kgBWi.p during gestation day 7 and then Curcumin therapy 300mg/kgBW were administered p.o ten days consecutively. Gestation day 17 all mice were sacrificed for histopathological examination.

Examination Procedure and Data Analysis:

The sample that had been obtained then fixated in formalin buffer and then stained with Hematoxylin and Eosin (HE). Liver histopathology examination was done with 400x magnification in various field of views with Nikon Eclipse Ci microscope. The scoring and grading model was used in this research refers to Arsad¹³: (0) showing no changes, and (1) (2) and (3)indicating mild, moderate and severe changes, respectively, while the grading was determined by percentage as follows: Changes less than 30% (<30%) showing mild changes, changes less than 30% – 50%,(<30% – 50%) indicating moderate changes and changes more than 50% (>50%) showing severe changes.

RESULT:

The statistic analytical were using Kruskal Wallis test and continued with Mann Whitney U, the result showed that T1 had numerous histopathological changes than the other groups 8, 80±1, 643; p< 0.01 then followed by T2 group 6, 60±1, 517; p< 0.01. T1 as the negative control group showed slightest number of histopathological changes with 1, 40 ± 0, 894; p< 0.01 and followed by T3 1, 60±1, 140; p< 0.01. The result showed that the number of histopathological changes is decreased in T3 group with 300mg/kgBW dose.

Figure 2.

The microscopic figure of liver (Hematoxylin-Eosin staining: 400x magnificent; Olympus CX21 microscope). Figure T0 showed normal centralist vein with radial structured, figure T1 showed the damage of liverparenchym that intoxicated with Cadmium, the structure of liver was changing and there was an accumulation of erythrocytes in sinusoid. Black arrow showed necrotic hepatocyte. Group T2 showed the healing of hepatocyte structure and the decreasing of necrotic hepatocyte. Groups T3 showed radial structured of hepatocyte and clear cytoplasm with normal centralist vein. Black arrow shows normal sinusoid and there was no erythrocyte accumulation

DISCUSSION:

The liver is the main target organ of Cadmium. Almost all Cadmium is absorbed and accumulated in liver¹³. The recent research was done to know the effect of Cadmium administration in liver showed apoptotic and necrotic on hepatocyte¹⁵. The damage caused by Cadmium generally is because the oxidative stress that binds to SH group and competitive to calcium^{16, 2, 17}.

Cadmium cause necrotic on central vein area, structure changing from hepatocyte plates that surroundcentral vein, also there is infiltration of the inflammatory cell around central vein and sinusoid beside fatty acid that also can be found, focal necrotic, picnotic on the nucleus, karyolysis, and cell Kupfer proliferation. The similar result found in recent study^2,18,19. Administration of Cadmium parenterally causes accumulation of Cadmium rapidly in the liver. Recent study shows that acute exposure of Cadmium causes hepatocyte necrotic. Generally necrotic of hepatocyte characterized by its centrolobular and expand to all livers’ lobus². Research done by El-Refaiy and Eissa, 2013 shows a liver histopathological figure with Mallory Azan staining, there was increasing collagen fiber in group that given Cadmium in zone I.

Histopathology examination from group that given Curcumin (T2 and T3) showed there was decreasing of livers’ damage and it was significant compare with group that given Cadmium 4.5 mg/kgBW (T1). Curcumin with dosage 150 mg/kgBW6, 60±1,517; p< 0.01 (T2) showed the decreasing of livers’ damage compare to T1that given only Cadmium 8, 80 ±1,643; p< 0.01.K3 group with dosage of Curcumin 300 mg/kgBW1, 60 ± 1,140; p< 0.01showed decreasing of livers’ damage and was significant nearly the same as control negative group (T0)1,40 ± 0,894; p< 0.01. This is indicated that Curcumin can decrease the number of cell death in liver.

Research done in rodent and in vitro model, Curcumin showed the potency as protective agent in some cases like nephrotoxicity^20,
21, immunotoxicity^{22, 23}, pulmonary disease²⁴, intoxicity in reproductive track^25-28, neurotoxicity²⁹, intoxi city in colon³⁰ and hepatotoxicity. The ability of Curcumin in decreasing livers’ damage is by decreasing livers’ inflammation, decreasing the effect of oxidative stress³¹, inhibit the activation of stellate cell^32,33 and support the function of mitochondria. Curcumin shows protective effect in molecular livers’ damage by regulating Keap1/Nrf2/ARE pathway³⁴.

CONCLUSION:

The conclusion is the administration of Curcumin after treated with Cadmium can decrease the number of cell death and improve the quality of liver histopathology.

CONFLICT OF INTEREST:

The author declared that there is no conflict of interest regarding the publication of this article.

REFERENCES:

1. Jarup L, Hellstorm L, and Alven T. Low Level Exposure to Cadmium and Early Kidney damage: the OSCAR Study. Occupational and Environmental Medicine. 2000; 57: 668-672.

2. Sinha M, Manna P and Parames CS. Induction of Necrosis in Cadmium-induced Hepatic Oxidative stress and Its Prevention by The Prophylactic Properties of Taurine. J. Trace Elements in Medicine and Biology. 2009; 23(4): 300-313.

3. Matović, V, Buha A, Dukic´-C´osicD and Bulat Z. Insight into the Oxidative Stress Induced by Lead and/or Cadmium in blood, Liver and Kidney. Elsevier. Food and Chemical Toxicology. 2015; 78: 130-140.

4. Matovic´ V, Dukic´-C´osic´ D, Buha A, and Bulat Z. Route Dose and Duration of Exposure to Cadmium-relevance to Oxidative Stress Induction. In: Bogaert, L., Coppens, N. (Eds.), Peroxidases: Biochemical Characteristics, Functions and Potential Applications. Nova Science Publishers, Inc, New York, 2013. 159–175.

5. Casalino E, Sblano C, Calzaretti G, and Landriscina C. Acute Cadmium Intoxication Induces Alpha-class Glutathione S-transferase Protein Synthesis and Enzym Activity in Rat Liver. Toxicology. 2006; 217: 240-245.

6. Liu L, Tao R, Huang J, He X, QuL, Jin Y, Zhang S and Fu Z. Hepatic oxidative stress and inflammatory responses with cadmium exposure in male mice. Environmental Toxicology and Pharmacology. 2015; 39: 229-236.

7. Garcia-Nìño WR and Chaverrì JP. Protection Effect of Curcumin Against Heavy Metals-induced Liver Damage. Elsevier. Food and Chemical Toxicology. 2014; 69: 182–201.

8. Lobo V, Patil A, Phatak A and Chandra N. Free radicals, antioxidants and functional foods: Impact on human health. Pharmacognosy Review. 2010; 4: 118–126.

9. Saeidnia S and Abdollahi M. Antioxidants: friends or foe in prevention or treatment of cancer: the debate of the century. Toxicology and Applied Pharmacology. 2013; 271:49–63.

10. Tapia E, Zatarain-Barron Z, Hernandez-Pando R, Zarco-Marquez G, Molina-Jijon E, Cristobal-Garcia M, Santamaria J, Pedraza-Chaverri J. Curcumin reverses glomerular hemodynamic alterations and oxidant stress in 5/6 nephrectomized rats. Phytomedicine. 2013; 20: 359–366.

11. Anthony B, Merina B, Iyer VS, Judy N, Lennert K, Joyal S. Apilot-cross Over Study to Evaluate Human Oral bioavability of BCM-95 (biocurcumax^TM); a Novel Bioenhanced Preparation of Curcumin. Indian Journal Pharmaceutical Sciences. 2008; 70(4): 445-449.

12. Dutta S, Padhye S, Priyadarsini KI, Newton C. Antioxidant and antiproliferative activity of Curcumine semicarbazone. Bioorganic and Medicinal Chemistry Letters. 2005; 15(11): 2738-2744.

13. Černý D, Lekić N, Váňová K, Muchová L, Hořínek A, Kmoníčková E, Zídek Z, Kameníková L, Farghali H. Hepatoprotective effect of Curcuminin lipopolysaccharide/D-galactosamine model of liver injury in rats: Relationship to HO-1/CO antioxidant system. Fitoterapia. 2011; 82: 786-791.

14. Arsad SS, Norhaizan ME and Hazilawati H. Histopathologic Changes in Liver and Kidney Tissues from Male Sprague Dawley Rats Treated with RhaphidophoraDecursiva (Roxb.) Schott Extract. J CytolHistol. 2014; 54: 1-6

15. DelRaso NJ, Foy BD, Gearhart JM, and Frazier JM. Cadmium Uptake Kinetics in Rat Hepatocytes: Correction for Albumin Binding. Toxicological Sciences. 2003; 26(1): 19-30.

16. Obianime AW and Roberts II. Antioxidants, Cadmium-Induced Toxicity, Serum Biochemical and the Histological Abnormalities of the Kidney and The Testes of The Male Wistar Rats. Nigerian Journal of Physiological Sciences. 2009; 24(2): 177-185.

17. Wang B, Li Y, Shao C and Tan Y. Cadmium and Its Epigenetic Effects. Current Medical Chemistry. 2012; 19: 2611-2620.

18. Klaassen CD, Liu J and Choudhuri S. Metallothionein: an Intacellular Protein to Protect Against Cadmium Toxicity. Annu Rev Pharmacol Toxicol. 1999; 39: 267-295.

19. Ersan Y, Ari I, and Koc E. Effect of Cadmium Compounds (Cadmium Para Hydroxybenzoate and Cadmium Chloride) on The Liver Mature Mice. Turkish Journal of Zoology. 2008; 32(2): 115-119.

20. Gong P, Chen FX, Ma GF, Feng Y, Zhao QY and Wang R. Endomorphin I Effectively Protects Cadmium Chloride-Induced Hepatic Damage in Mice. Toxicology. 2008; 251: 35-44.

21. Tarasub N, Tarasub C, Devakul W, and Ayutthaya N. Protective role of Curcumin on Cadmium-induced nephrotoxicity in rats. Jurnal Environmental Chemical Ecotoxicology. 2011; 3:17–24.

22. Deevika B, Asha S, Taju G, and Nalini T. Cadmium acetate induced nephrotoxicity and protective role of Curcumin in rats. Asian Journal Pharmacology Clinical. 2012; 5: 186–188.

23. Pathak N and Khandelwal S. Comparative efficacy of piperine, Curcumin and picroliv against Cdimmunotoxicity in mice. Biometals 2008; 21: 649–661.

24. Alghasham A, Ta S, and Ar M. Effect of Cadmium-polluted water on plasma levels of tumor necrosis factor-a, interleukin-6 and oxidative status biomarkers in rats: protective effect of Curcumin. Food Chemical Toxicology. 2013;59: 160–164.

25. Rennolds J, Malireddy S, Hassan F, Tridandapani S, Parinandi N, Boyaka P and Cormet-Boyaka E.. Curcumin regulates airway epithelial cell cytokine responses to the pollutant Cadmium. Biochemical Biophysical Research. Communication. 2012; 417: 256–261.

26. Souza V, Bucio L, Jay D, Chavez E, Gutierrez-Ruiz M.C. Effect of Cadmium on calcium transport in a human fetal hepatic cell line (WRL-68 cells). Toxicology 1996; 112: 97–104.

27. Salama A and El-Bahr S. Effect of Curcumin on Cadmium-induced oxidative testicular damage in rats. Journal Medicine Research Institute. 2007; 28: 167–173.

28. Oguzturk H, Ciftci O, Aydin M, Timurkaan N, Beytur A, and Yilmaz F. Ameliorative effects of Curcumin against acute Cadmium toxicity on male reproductive system in rats. Andrologia 2012; 44: 243–249.

29. Singh P, Deora K, Sankhla V and Mogra P. Curcumin rendered protection against Cadmium chloride induced testicular damage in Swiss albino mice. Journal Cell Molecular Biology. 2012; 10: 31–38.

30. Daniel S, Limson JL, Dairam A, Watkins GM andDaya S. Through metal binding, Curcumin protects against lead- and Cadmium-induced lipid peroxidation in rat brain homogenates and against lead-induced tissue damage in rat brain. J. Inorg. Biochem. 2004; 98: 266–275.

31. Mathuria N and Verma R. Curcumin ameliorates aflatoxin-induced lipid peroxidation in liver, kidney and testis of mice – an in vitro study. Acta Pol. Pharm. 2007; 63, 413–416.

32. Zheng S, Yumei F and Chen A. De novo synthesis of glutathione is a prerequisite for Curcumin to inhibit hepatic stellate cell (HSC) activation. Free Radic. Biol. Med. 2007; 43: 444–453.

33. Priya S and Sudhakaran P. Curcumin-induced recovery from hepatic injury involves induction of apoptosis of activated hepatic stellate cells. Indian J. Biochem. Biophys. 2008; 45: 317–325.

34. Subudhi U, Das K, Paital B, Bhanja S and Chainy G. Alleviation of enhanced oxidative stress and oxygen consumption of L-thyroxine induced hyperthyroid rat liver mitochondria by vitamin E and Curcumin. Chem. Biol. Interact. 2008; 173: 105–114.

Received on 20.11.2017 Modified on 20.12.2017

Research J. Pharm. and Tech. 2018; 11(3): 863-866.

DOI: 10.5958/0974-360X.2018.00160.9