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EVALUATION OF THE IMPACT OF CADMIUM CHLORIDE AND LEAD ACETATE ON THE LUNG TISSUE OF ALBINO FEMALE RATS BY PHYSIOLOGICAL AND HISTOLOGICAL METHODS
Q.A. Razooqi , I.S. Dalas, E.M. Harz
College of Education for Pure Sciences, University of Tikrit, Tikrit/Iraq. * Corresponding author: [email protected], [email protected]
Abstract. This study examined female rats' physiological and histological responses to cadmium chloride and lead acetate. Histological lung tissue examinations included oxidative stress and antioxidant status. Six female rat groups were studied. A group that served as a control was provided with water that had been distilled. The dosage of cadmium chloride that was administered to the second group was 5 mg/kg, whereas the dosage that was administered to the third group was 10 mg/kg. The fourth group received a dosage of lead acetate that was 50 mg/kg, whereas the fifth group received 100 mg/kg. All of the standard concentrations of lead acetate and cadmium chloride were administered to the sixth group in accordance with their protocol. Following thirty days of treatment of cadmium chloride and lead acetate to rats, the levels of oxidative indicators like MDA and 8-OHDG showed a substantial increase (P < 0.05). On the other hand, the levels of antioxidants like GSH, SOD, and CAT showed a significant drop (P < 0.05 following the administration of these substances. Histological research has shown that exposure to cadmium chloride and lead acetate is associated with an increased risk of blood clots in the lungs as well as a thickening of the pulmonary alveolar wall. This is in comparison to a control group. These results demonstrate that cadmium chloride and lead acetate treatment adversely affected lung tissue's physiological and histological properties. The researchers discovered that the detrimental effect was more pronounced when the two drugs were administered concurrently to rats.
Keywords: lead acetate, cadmium chloride, 8-OHDG, lung tissue.
List of Abbreviations
CdCl2 - Cadmium chloride
Pb(C2H3O2)2 - Lead acetate
8-OHDG - 8-hydroxy-2' -deoxyguanosine
MDA - Malondialdehyde
GSH - Glutathione
SOD - Superoxide dismutase
CAT - Catalase enzyme
Introduction
Lead and cadmium are significant heavy metals prevalent in contaminated ecosystems. Both natural and anthropogenic sources, such as metal mining, smelting, and various chemical products, expose humans and animals to pollutants. These contaminants are prevalent in the environment, resulting in reciprocal detriment to living organisms and additional adverse effects on human health (Sultan et al, 2018). Heavy metals and their buildup in the environment increased due to the nineteenth-century industrial revolution, which gradually increased heavy metal use in industry. Due to their strong environmental resistance, these metals rapidly infiltrate the food
chain (Barn et al., 2019; Kumar & Sharma, 2019). Due to their lengthy biological half-life, sluggish metabolic rate, and limited excretion, these heavy metals can accumulate in the placenta, small intestine, liver, and kidneys. Heavy metal accumulation can poison these organs. (Xiong, 2020; Zhu, 2021; Mohamed, 2022). Cadmium (Cd) is a heavy metal found in nature as CdCl2, CdS, CdS, and CdO (Janicka et al, 2014). Cadmium is found in water, soil, tobacco smoke, and numerous businesses like battery manufacture. Cadmium combines with calcium, zinc, and iron in the body to limit their efficiency (Jain, 2020). Cadmium indirectly forms free oxygen radicals in the cell, damaging mitochondrial electron transport chain sites. Free radicals alter cell DNA, proteins, lipids, and carbohydrates. Some organs may develop morphological problems from this condition (Fouad & Jresat, 2015; Zhang et al., 2021). Many sectors use lead, including lead-acid batteries and colouring additives. Lead is hazardous and harms bodily and fetal organs (Zhang et al., 2020; Kabamba & Tuakuila, 2020). Continuous lead poisoning can cause lead buildup and
death (Nkwunonwo et al., 2020). Numerous studies show lead metal damages humans and animals. Male sexual drive, sperm production, quality, hormone synthesis, and management are affected (Pizent et al., 2012). Another study found lead damages the hypothalamic axis, pituitary gland, and testicles. In rats, lead acetate lowers antioxidant enzyme activity in the male testicles, affecting steroid synthesis and sperm parameters (Oyeyemi et al., 2019). Lead acetate in albino rats' drinking water for 45 days reduced testicular weight, testosterone, and sperm. The medication lowered antioxidant enzymes and raised MDA, indicating oxidative stress (Hassan et al., 2019).
Materials and Methods
Each of six female rat groups had five similar-weight rats. The rats were randomly assigned to groups. After two weeks of acclimatization, the animals received the following doses: The experiment's control group received 30 days of distilled water. Five milligrams per kilogram of body weight of cadmium chloride was given to the second group. 30 days were spent on this dose (AL-Kraie et al., 2020). The third group received double the permitted dose of cadmium chloride (10 mg/kg body weight) for 30 days gave the fourth group 50 mg of lead acetate per kg of body weight for 30 days. The fifth group received 100 mg of double lead acetate per kg of body weight for 30 days. The sixth group received a dual-dose immunization of five and fifty milligrams per kilogram of cadmium chloride and lead acetate. This immunization lasted 30 days.
Determination of the doses of pollutants
The doses of cadmium chloride and the usual dose of lead acetate were determined based on previous studies, while the doses were doubled to clarify that exposure to more pollutants because double negative toxic effects or leads to the death of animals.
Preparing the studied samples
A) Blood serum samples
Upon conclusion of the experiment, the mated female rats were euthanized to procure blood samples. Blood samples were collected and processed to isolate serum for biochemical analysis of oxidative components.
B) Histological preparation
Subsequent to the extraction of the lung tissue, it was rinsed with water, fixed in 10% formalin for twenty-four hours, dehydrated using ascending concentrations (70%, 80%, 95%, 100%, 100%), cleared with xylene, and ultimately embedded in paraffin. All of these procedures were conducted subsequent to the dissection of the lung tissue. Paraffin-fixed lung tissue sections were sliced to a thickness of five millimeters and subsequently stained with hae-matoxylin and eosin. An optical microscope with a magnification of 400x was employed to examine the samples (Anthony, 2016).
Statistical Analysis
The statistical analysis was carried out by means of the analysis of variance (ANOVA) test, and the Duncan test was utilized to evaluate the significance of the findings while maintaining a significance level of (P < 0.05). The year 1989, Lars and World.
Ethical approval
We conducted the research was performed in accordance with the minutes from the Department of Biology and the Ethical Committee of Tikrit University, Iraq, College of Education for Pure Sciences, Department of Biology, during the first session on 16/9/2024.
Results
The physiological indicators, specifically oxidative stress markers (MDA, 8-OHDG), went up significantly (P < 0.05) in females that were exposed to cadmium chloride and lead acetate (Fig. 1, 2). On the other hand, antioxidants showed a significant decrease in their levels (Fig. 3, 4, 5) (GSH, SOD, and CAT) and demonstrated a noteworthy decrease (P < 0.05) in comparison to the control group. As shown in Figures 8, 9, 10, 11, 12, 13, 14, and 15, the administration of varying doses of cadmium chloride and lead acetate to rats resulted in significant histological damage to lung tissue. Blood congestion, thickening of pulmonary alveolar walls, and an increased presence of inflammatory cells characterized this damage. We observed this damage in comparison to the control (Fig. 6, 7).
Fig. 1. Different doses of cadmium chloride and lead acetate given to rats led to a significant increase (P < 0.05) in the level of Malondialdehyde (MDA) compared to the control group
Fig. 2. Different doses of cadmium chloride and lead acetate administered to rats led to a significant increase (P < 0.05) in the level of 8-hydroxydeoxy-guanosine (8-OHdG) compared to the control group
Fig. 3. Dosing rats with different doses of cadmium chloride and lead acetate resulted in a significant decrease (P < 0.05) in the level of superoxide dis-mutase (SOD) compared with the control group
Fig. 4. Giving rats different doses of cadmium chloride and lead acetate led to a significant decrease (P < 0.05) in the catalase enzyme (CAT) compared to the control group
Fig. 5. Giving rats different doses of cadmium chloride and lead acetate led to a significant decrease (P < 0.05) in the level of glutathione (GSH) compared to the control group
Fig. 6. A lung segment from the control group, illustrating the alveoli (Alv), pulmonary sacs (AVS), and bronchioles (Br) in their typical morphology. H & E 100X
Fig. 7. Lung section of the control group showing the alveoli (Alv) and bronchioles (Br) in their normal shape. H & E 400X
Fig. 9. The group's lung, which received low-dose cadmium chloride treatment, displays normal blood vessel (CON) and bronchial tube (Br) congestion. H & E 400X
Fig. 11. A lung slice from the group that received low-dose lead acetate treatment revealed fibrosis (Fb), an accumulation of inflammatory cells (IF), and thickening of the blood vessel walls (TW). H & E 100X
Fig. 8. A normal-appearing cross-section of the bronchial tubes (Br) and congestion of blood vessels (CON) in the group that received low-dose cadmium chloride in their lungs. H & E 100X
Fig. 10. A section of the lung from the group that was treated with low-dose cadmium chloride demonstrates the thickening of the alveolar walls (TW) and the congestion of blood vessels (CON). H & E 400X
Fig. 12. Blood congestion (CON) was observed in the lung region of the group that received low-dose lead acetate treatment. H & E 400X
Fig. 13. A piece of the lungs from the people who were given high doses of lead acetate shows that the walls of the blood vessels have thickened (TW) and that inflammatory cells have gathered (IF), along with crowding (CON). H & E 100X
Fig. 14. Fibrosis inside the lung tissue (Fb) and thickening of the blood vessel walls (TW) in the group that was treated with high-dose lead acetate (G). H & E 400X
Fig. 15. The low-dose group treated with cadmium chloride and lead acetate had extensive blood hem-orrhaged within their lung tissue, as shown in this part of their lung (H). H & E 400X
Discussion
An organism's tissues absorb, distribute, and accumulate heavy metals based on several factors. The properties of the metals, their forms, their route of exposure, their ability to bind to cell ligands, and the species' sensitivity are among these factors. Both cadmium and lead enter the bloodstream after absorption in the intestines (Swiergosz-Kowalewska, 2001; Tim-chalk et al., 2006). According to Mladenovi'c et al. (2014), the increase in the concentration of Malondialdehyde (MDA) may be due to the formation of free radicals and their attack on the saturated fatty acids present in the cell membranes, which leads to the formation of fatty acid peroxidation, causing a loss of function of the cell membranes due to the loss of their bio-fluidity and thus cell death (Demirkol et al., 2012). Cadmium chloride, on the other hand, is a pollutant that causes the production of free radicals, which reduce the efficiency of antiox-idants, and thus cause an imbalance between the level of antioxidants and free radicals, which causes an increase in the oxidative state (Alfar-hani et al., 2021). Metal exposure leads to oxidative stress, which in turn produces free radicals and lipid peroxidation at varying concentrations of cadmium chloride. Further, lead acetate can obstruct metabolism by binding to the -SH groups of numerous proteins, including enzymes. As a result, the activity of SOD and CAT decreases, leading to a drop in their levels. This is supported by studies conducted by Ma-tovic et al. (2015) and Flora et al. (2012). It not only leads to an increase in red blood cells and a decrease in glutathione levels, but also generates a significant amount of MDA and H2O2. Omobowale et al. conducted the study in 2014. The study's results showed that the group that was given low doses of chloride and lead acetate had a worse reaction. If it causes a clear drop in antioxidants and a significant rise in the levels of oxidation products (8-OHDG and MDA), along with clear histological damage in lung tissue compared to the other groups, then these results are in line with what Erel (2004) and Andjelkovic et al. (2019) say. The role of cadmium is that it produces a large amount of free radicals that work for the oxidative degra-
dation of lipoprotein and DNA; cadmium stimulates apoptosis and/or cell necrosis. It creates reactive oxygen species like superoxide radicals, hydroxyl ions, and hydrogen peroxide (EL-Refaiy & Eissa, 2013; Al-Derawi, 2018). Lead is a very toxic pollutant, and there are many signs of poisoning that appear on the body due to exposure to this pollutant, including intestinal and neurological signs. This depends on the period and amount of exposure to this substance, which leads to both acute tox-icity as well as chronic toxicity (Alomran & Shleamoon, 1988; Schwartz, 2001). Many studies have confirmed that exposure to lead accumulates in the kidney cortex and marrow. In rats dosed with lead, in addition, it was found that exposure to lead causes more toxic effects on various organs of the body. For example, it causes an effect on the kidneys, lungs, bones, liver, heart, and blood, and finally the toxic effect may reach the testicles and the brain (Aziz et al., 2012). In addition, chronic exposure to lead leads to the appearance of inflammatory cells in the tissues of the body. Lead's interaction with bodily tissues'
enzymes and proteins may weaken the antioxidant defense mechanism, triggering a traditional inflammatory response through the generation of reactive oxygen species (ROS). Recent studies have indicated that the cause of pathophysiological changes in body tissues, including the liver, for example, may be due to oxidative stress or programmed cell death (Fortoul et al., 2004).
Conclusion
This study concludes that when animals receive combined doses of lead acetate and cadmium chloride, the effects are more pronounced and lead acetate has a more detrimental effect on lung tissue than cadmium chloride, both physiologically and histologically.
Conflict of interest
The authors declare no conflict of interest.
Acknowledgments
We extend our thanks and appreciation to the workers in the animal house of the College of Veterinary Medicine at Tikrit University.
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