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 Table of Contents  
ORIGINAL ARTICLE
Year : 2020  |  Volume : 4  |  Issue : 1  |  Page : 52-58

Biochemical and histological evaluation of testicular and hepatic acute toxicity induced by formaldehyde and the possible protective effect of curcumin in adult male albino rats


Department of Forensic Medicine & Clinical Toxicology, Faculty of Medicine for Girls, Al-Azhar University, Cairo, Egypt

Date of Submission25-Dec-2019
Date of Decision06-Jan-2020
Date of Acceptance08-Jan-2020
Date of Web Publication20-Apr-2020

Correspondence Address:
Associate Professor Wesam Abdelsalam Abdelwahab
Associate Professor at Forensic Medicine and Clinical Toxicology Department, Faculty of Medicine for Girls, Al-Azhar University
Egypt
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/sjamf.sjamf_111_19

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  Abstract 


Background and aim Formaldehyde (FA) is an organic carbon compound with adverse health effects by inducing oxidative stress that affect multiple tissues of the exposed animals such as liver, brain, and gonads. Curcumin has desirable preventive therapeutic properties owing to its antioxidant and anti-inflammatory properties. This study aimed to evaluate toxic effects of FA on liver and testes of the adult male albino rats and assess the probable protective role of curcumin against these effects through biochemical and histological studies.
Methodology Twenty-four adult male albino rats were divided into four groups: control, curcumin (100 mg/kg/day by intragastric tube), FA (10 mg/kg/day intraperitoneally), and FA+curcumin, daily for 2 weeks. Blood samples, livers, and testes were obtained for biochemical and histological evaluation.
Results FA resulted in elevation of aspartate aminotransferase, alanine aminotransferase, and total bilirubin and decrease in albumin and testosterone levels and alterations in the normal hepatic and testicular structures. Treatment with curcumin improved these changes.
Conclusion FA administration damages and deteriorates hepatic and testicular structures and functions, and curcumin improved that, which might be attributed to its antioxidative and anti-inflammatory properties.
Recommendation It is recommended to use curcumin as a protective agent in FA toxicity.

Keywords: formaldehyde, hepatic, testicular toxicity, curcumin


How to cite this article:
Abdelwahab WA, Megahed RM. Biochemical and histological evaluation of testicular and hepatic acute toxicity induced by formaldehyde and the possible protective effect of curcumin in adult male albino rats. Sci J Al-Azhar Med Fac Girls 2020;4:52-8

How to cite this URL:
Abdelwahab WA, Megahed RM. Biochemical and histological evaluation of testicular and hepatic acute toxicity induced by formaldehyde and the possible protective effect of curcumin in adult male albino rats. Sci J Al-Azhar Med Fac Girls [serial online] 2020 [cited 2020 May 30];4:52-8. Available from: http://www.sjamf.eg.net/text.asp?2020/4/1/52/282861




  Introduction Top


Formaldehyde (FA) is highly water-soluble aldehyde ingested in fresh water, food, and drugs [1]. All civilized population are exposed to FA, as it is part of general outdoor and indoor environment [2].

FA is found in commonly used products such as cosmetics and cleaning agents [3] produced from burning of tobacco [4] and used in storage of cadavers, tissue fixation in microscopic laboratories, and as dental coating materials. Moreover, formalin is found in dialysis solutions [5].

FA is present in the manufacturing process of some vaccines such as diphtheria, polio, and tetanus preparations, so infants can be exposed to its toxicity [6]. Several drugs that are used in the treatment of cancers are formulated with FA [7]. Some hair smoothing products contain FA [8].

FA induces oxidative stress, leading to toxicity in multiple tissues such as liver and gonads [9]. It has high ability for reacting with DNA, RNA, and protein, which leads to many health hazard [10].

FA increases cell apoptosis in the liver [11], affects cytochrome P450 [12], declines serum testosterone level, and increases infertility in men [13].

Curcumin has multiple biological and pharmacological effects including antioxidant and anti-inflammatory [14]. It binds to free radicals, so protects DNA from oxidative damage [15].

This study aimed to evaluate toxic effects of FA on liver and testes of adult male albino rats and to assess the probable protective role of curcumin against these effects through biochemical and histological studies.


  Methodology Top


Twenty-four adult male albino rats (obtained from Helwan animal breeding farm, Cairo, Egypt.), weighing 150–200 g were used in this study. They were maintained in stainless steel cages in a well-ventilated animal house at normal temperature (22±5°C) under a 12 : 12-h light–dark cycle. They were fed normal feeding and given water. They were kept under suitable conditions for one week for adaptation before the start of the experiment.

Handling of animals was done following the rules for the experimental research ethics, approved by Research Ethics Committee at faculty of Medicine for Girls Al-Azhar University, Egypt, and conforms with the Guide for the Care and Use of Laboratory Animals [16].

Tested substances

FA was in liquid form of 40% concentration. Curcumin powder was dissolved in distilled water before administration to the rat. They were obtained from El Gomhouria Company for Chemical and Medical Trading, Cairo, Egypt.

Experimental design

Rats were divided equally into four groups: control (rats were received normal feeding and distilled water only), curcumin (rats were received curcumin 100 mg/kg/day by intragastric tube) daily for 2 weeks [17]; FA (rats were administered FA 10 mg/kg/day intraperitoneally daily for 2 weeks) [17], and FA+curcumin (rats were received them both daily for 2 weeks).

Serum and tissue collection

At the end of experiment, blood samples were taken from the medial canthus of the eyes of rats using glass capillaries while they were under anesthesia by diethyl ether. The samples were collected in clean dry test tubes and left at room temperature for 20 min to clot. Blood samples were centrifuged at 4000 rpm for 15 min to separate the sera. The sera were then stored at −20°C for biochemical analysis.

Then all animals were killed by decapitation while they were under anesthesia by diethyl ether inhalation. Both testes and livers were dissected and utilized for histological study by hematoxylin–eosin stain. Testis was weighed before histological study.

Biochemical evaluation

  1. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were assessed according to studies by IFCC [18],[19], respectively.
  2. Albumin was assessed according to Doumas et al. [20].
  3. Total bilirubin was assessed according to Walter and Gerald [21].
  4. Testosterone serum level was measured by enzyme-linked immunosorbent assay kit (Diaplus, USA; Diagnostic System Laboratories Inc., STAT FAX, Webster, Texas, USA) as recommended by the manufacturer.


Histological study

Livers and testes specimens were prepared for staining with hematoxylin–eosin stain according to Kieranan [22].

Statistical analysis

Data were statistically expressed as means±SD and compared using the one-way analysis of variance followed by Tukey’s post-hoc test. Level of P value less than 0.05 is used as the criterion of significance. Statistical analysis was performed using the Statistical Package for the Social Sciences, version 22, for Windows (SPSS Inc., California, USA).


  Results Top


Biochemical evaluation

Regarding liver

FA significantly increased levels of ALT, AST, and bilirubin. However, albumin level was significantly decreased as compared with control rats ([Table 1]). Administration of curcumin with FA showed significant decrease in serum levels of ALT, AST, and bilirubin and significant increase in albumin level as compared with FA group.
Table 1 Comparative analysis of mean values of aspartate aminotransferase, alanine aminotransferase, albumin, and total bilirubin in all studied groups

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Regarding testes

FA induced a significant decrease in testosterone hormone level and testes weight, but after co-administration of curcumin, testosterone hormone level and testes weight significantly increased when compared with FA group ([Table 2]).
Table 2 Comparative analysis of mean values of testis weight and testosterone hormone level in all studied groups

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Histological examination

Liver

Liver in control and curcumin group revealed normal hepatic architecture ([Figure 1]). FA induced disruption in the normal hepatic architecture ([Figure 2]). Co-administration of curcumin with FA improved most of these changes ([Figure 3]).
Figure 1 Liver section from control displayed (a) normal hepatic cords (HC) around central vein (CV), separated by blood sinusoids (S), flattened endothelial lining of central vein (arrow head), and polyhedral eosinophilic hepatocytes with central rounded vesicular one (white arrows) or two nuclei (black arrow). (b) Portal triad (P) at corner of hepatic lobule branch of hepatic artery, portal vein, and bile duct (BD) lined with cubical epithelial cells. Curcumin group (c–d) normal hepatic pattern as control. Hematoxylin–eosin stain, ×400.

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Figure 2 Liver section of formaldehyde group displayed congested central vein (CV), congested blood sinusoids (S), and widespread vacuolated and ballooned hepatocytes. Most hepatocytes have multiple intracytoplasmic vacuoles (thin black arrows) and small deeply stained pyknotic nuclei (white arrow head). Some vacuoles coalesce with each other (thick black arrows). (b) Dilated and congested portal vein (PV) with mononuclear cellular infiltration (curved arrows) and dilated bile ductule (BD) at the portal tract area (P). Hematoxylin–eosin stain, ×400.

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Figure 3 Liver section of formaldehyde+curcumin group displayed (a) preservation of hepatic architecture, hepatic cords (HC) arrangement around the central vein (CV) separated by blood sinusoids (S), polyhedral eosinophilic hepatocytes with central rounded vesicular one (white arrows) or two nuclei (black arrow), few hepatocytes with multiple intracytoplasmic vacuoles of different sizes and centrally located rounded nuclei. (b) Portal triad (P) showing congested branch of portal vein (PV) and normal bile duct (BD) is lined with cubical epithelial cells. Hematoxylin–eosin stain, ×400.

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Testis

Testis in control and curcumin group revealed normal architecture ([Figure 4]). FA induced disruption in the normal testicular architecture ([Figure 5]). Co-administration of curcumin with FA improved most of these changes ([Figure 6]).
Figure 4 Testicular tissue sections of control: (a) normal structure of mature closely packed seminiferous tubules (ST), narrow interstitial spaces (IS) contained interstitial tissue in between ST. (b) ST surrounded by thin regular basement membrane (thin arrow), lined by  Sertoli cells More Details(thick arrows), normal spermatogenic series, including spermatogonia (Sg), primary spermatocytes, spermatids (Sp), spermatozoa (S)well-developed IS contains clusters of Leydig cells (L) in between tubules. Curcumin group (c–d): normal pattern appears as control (hematoxylin–eosin stain, ×100 (a, c), ×400 (b, d).

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Figure 5 Testicular tissue sections of formaldehyde group: (a, b) disrupted tubules with loss of their normal architecture, widening of IS with areas of hemorrhage (H). (b) Vacuoles (V) and cellular debris (arrow) in lumen of some ST. (c) Vacuoles and hyaline material (Hy) observed in lumen of tubule. (d) Vacuoles, cellular debris of exfoliated cells (arrows) separated from their basement membrane observed in lumen of tubule. Pyknotic interstitial L around congested blood vessels (BV) in IS (hematoxylin–eosin stain), ×100 (a, c), ×400 (c, d).

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Figure 6 Testicular tissue sections from formaldehyde+curcumin-treated group. Seminiferous tubules (ST) are surrounded by thin regular basement membrane (thin arrow), lined by Sertoli cells (thick arrows), and normal spermatogenic series, including spermatogonia (Sg), primary spermatocytes (PS), spermatids (Sp), spermatozoa (S). Well-developed interstitial tissue (IS) contains clusters of Leydig cells (L) in between the tubules (hematoxylin–eosin stain, ×100 (a), ×400 (b).

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  Discussion Top


FA has received increased attention as a pollutant with adverse health effects by inducing oxidative stress that affect multiple tissues of the exposed animals such as liver and gonads [23].

Lipid peroxidation and reactive oxygen species (ROS) are involved in FA-induced toxicity [24].

It has been focused on the protective effect of plant products or medicinal plants that having antioxidant properties in reducing free radical-induced tissue damage [25].

Curcumin, a yellow pigment from Curcuma longa, has desirable preventive therapeutic properties owing to its antioxidant and anti-inflammatory properties [26].

This study aimed to evaluate the toxic effects of FA on both testes and liver of the adult male albino rats and to assess the probable protective role of curcumin against these effects through biochemical and histological studies.

Results of the present study revealed that acute FA toxicity resulted in significant increase in serum levels of AST, ALT, and total bilirubin. However, albumin level significantly decreased as compared with the control group. Co-administration of curcumin with FA caused significant decrease in the serum levels of ALT, AST, and bilirubin and significant increase in albumin level when compared with the FA group.

These results were in agreement with Afrin et al. [27] and could be owing to the changes in the histological structure of the livers of the rats that were exposed to FA, as Kimbell et al. [28] found that FA caused cytotoxicity by reacting directly with tissue constituents.

In the present study, FA caused alternations of normal histological structure of the liver, which was improved by co-administration of curcumin. This was in agreement with Kotob et al. [29] and were supported by Gurel et al. [30], who stated that FA can cause severe tissue injury by producing ROS.

However, JiménezVillarreala et al. [31] found no significant changes in histological analysis of the liver between control group and FA group.

The current work revealed significant reduction in weight of testis in FA group in comparison with control group. Co-administration of curcumin with FA resulted in improvement in weight of testis in comparison with FA group.

These findings agreed with those of Askaripour et al. [32], whereas Zahra et al. [13] found no significant differences between FA and control group in testis weight.

Hegazy et al. [23] attributed testicular weight reduction in FA group to seminiferous tubule atrophy, as was the histological findings in the present study.

In the present study, FA resulted in significant decrease in testosterone hormone level but curcumin with FA increased its level.

This was in accordance with Askaripour et al. [32] and could be owing to induction of oxidative stress that affects male’s reproduction system [9].

Previous studies with FA showed significant changes in testis structure, sex hormone profile, and sperm parameters [33].

FA-induced oxidative stress, ROS generation, and methylation of DNA lead to chromosomal damage, fragmentation, protein cross-linking of DNA, and structural and functional alterations of enzymes, hormones, and proteins [31].Hegazy et al. [23] explained the decrease in the testosterone by the deformation and high lipid accumulation of Leydig cells after administration of FA that affects testosterone biosynthesis, as were found in the present study.

FA caused atrophy and degeneration of seminiferous tubules by increasing ROS production in many tissues including reproductive organs [34]. Excessive free radical production in testis may increase germ cell apoptosis [33].

FA in the present study disrupted normal histological structure of rat testis, but co-administration of curcumin improvement that. This was in agreement with Vosoughi et al. [35].

Inhibition of generation of free radicals or antioxidant activity plays an important role in protection against testicular toxicity and so curcumin succeeded in reducing free radical-induced tissue damage [36].

The anti-inflammatory effect of curcumin is owing to its ability to inhibit important enzymes that mediate inflammatory processes such as cyclooxygenase-2, lipoxygenase and inducible nitric oxide synthase [26].

Conclusion and Recommendation

FA caused damages in hepatic and the testicular structures and deteriorated their functions. Oral administration of curcumin showed protective effects and improved their functions and structures, which might be attributed to its antioxidative and anti-inflammatory properties. So, curcumin is recommended as a protective agent in FA toxicity.

Acknowledgements

The authors thanks Dr Mona Abd-Elgalil; Lecturer at Histology Department, Faculty of Medicine for Girls, Al-Azhar University for her assistance in histological examination.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6]
 
 
    Tables

  [Table 1], [Table 2]



 

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