The Effect of Pseudoephedrine (Sudafed) on Kinetic activity and histology of Livers and Kidneys in Albino Mice

Ban Jasim Mohamad¹, Faeza Aftan Zghair²

¹Biology Department, College of Science, University of Baghdad. Baghdad, Iraq.

²Department of Pathology and Forensic Medicine, College of Medicine, Al-Iraqia University, Baghdad, Iraq.

*Corresponding Author E-mail: ban.jasim@sc.uobaghdad.edu.iq. faizarawi@yahoo.co.uk

ABSTRACT:

Pseudoephedrine (PSE) or (Sudafed) is one of the sympathomimetic group of drugs (ephedrine, PSE and amphetamines) which effects cardiovascular system, respiratory system, and gastrointestinal tract. However, only little researches had supported its effect on solid abdominal organs. This study aims to investigate the effects of different doses of Sudafed in the liver and kidney of albino mice. The current study included 18 albino mice grouped into 2 groups: control (3 mice), and acute group (15 mice). The acute group was further subdivided into 5 subgroups, each subgroup of 3mice wasgiven a lonely intaperitonial injection of 0.3ml of the following conc. (500mg/kg, 250mg/kg, 125mg/kg, 62.52mg/kg, and 31.24mg/kg) for 24hrs. After the mentioned period, the mice of all subgroups were sacrificed and the livers and kidneys were removed, processed, sectioned and stained for histological analysis. Results of liver analysis using 500mg/Kg Sudafed intraperitoneallyshowed mild ballooning degeneration of hepatocytes and central vein congestion, while lower doses (250mg/Kg – 31.42mg/Kg) revealed less prominent effect or no significant pathological changes.Moreover, sections from the kidney with the 500mg/Kg Sudafed intraperitoneally showed mild hydropic swelling of tubular epithelium with congestion of intertubular blood vessels and relatively healthy glomeruli. Lower doses revealed no significant pathological changes. Conclusion: The present study demonstrated various pathological effects of PSE on kinetic activity, and histology of Livers and Kidneys of albino mice.

KEYWORDS: Pseudoephidrine, Sudafed, Toxic effect of PSE.

INTRODUCTION:

Pseudoephedrine (Sudafed) is one of the oldest molecules used in the treatment of nasal and sinus congestion or to address congestion in the Eustachian tubes of the ear¹, as well as to treat symptoms of rhinitis and rhinorrhea². It increases blood pressure, heart rate, respiratory rate, causes bronchial tree dilation, relaxation of the smooth muscles of the gastrointestinal tract, pupillary dilation, vasoconstriction of cutaneous blood vessels, vasodilation in the skeletal muscles, redistribute blood from the skin to the skeletal muscles and brain. These effects are more pronounced with higher doses and may cause complications during maximal exercise.

It also has a number of other uses, and can be administered as a stimulant or as a pharmaceutical agent that promotes wakefulness, and improvement running performance³and is included on the International Olympic Committee's list of banned substances⁴. This is mainly due to its chemical similarity to amphetamines and the assumption that (PSE) drug may be ergogenic in nature⁵. Oral form is generally considered to be the optimal administration route, as it has been evidenced to be less likely than topical application to cause rebound congestion⁶.

The salts pseudoephedrine hydrochloride and pseudoephedrine sulfate are found in many over-the-counter drugs either as single-ingredient preparations, or more commonly in combination with antihistamines, paracetamol (acetaminophen) and/or ibuprofen. Sudafed is a trademark for a common brand which contains pseudoephedrine hydrochloride. Drugs containing pseudoephedrine are commonly used as a treatment for Rhinitis (an inflammation of the mucous membranes in the nose). Unlike antihistamines, which modify the systemic histamine-mediated allergic response, pseudoephedrine only relieves nasal congestion commonly associated with colds or allergies⁷.

Pseudoephedrine acts directly on both alpha- and, to a lesser degree, beta-adrenergic receptors. Through direct action on alpha-adrenergic receptors in the mucosa of the respiratory tract, pseudoephedrine produces vasoconstriction. Pseudoephedrine relaxes bronchial smooth muscle by stimulating beta 2-adrenergic receptors. Like ephedrine, pseudoephedrine releasing norepinephrine from its storage sites, an indirect effect. This is its main and direct mechanism of action. The displaced noradrenaline is released into the neuronal synapse where it is free to activate the postsynaptic adrenergic receptors⁸.

In February 2014, the French Otorhinolaryngology Society, stated that vasoconstrictors “involve a risk of stroke and severe neurological effects” and that they “are often too risky for use against a simple cold”⁴. More common side effects include nerviness, restlessness and trouble in sleeping. While less common side effects are difficult or painful urination, dizziness or light-headedness, fast or pounding heartbeat, headache, increased sweating, nausea or vomiting, trembling, unusual paleness and weakness⁹. Overall, the review showed that the effect of pseudoephedrine is dose-dependent. None of the reviewed studies showed histologic effect on liver and kidney with different doses of the drug.

Aim of the study is to investigate the toxic effect of some doses of this drug on activity, behavior and histology of liver and kidney of albino mice.

EXPERIMENTAL DESIGN AND PROTOCOLS:

1. Experimental Animals:

Eighteen, 6-week-old male mice, weight 20-25gm were purchased from National Center for Drug Control and Research in Baghdad, and kept under standardized environmental conditions; constant temperature, moisture and with a 12-hr. light regime without stress factors. Mice were allowed to take laboratory food and water.

2. Design of the study:

Mice were randomized into two groups, and treatment was carried out according to the following groups:

A. Group I: Control Group; contains 3 mice, received standardized lab food and water without treatment.

B. Group II: Acute group; contains 15 mice, divided into 5 subgroups of 3 mice per each, received a single intra-peritoneal injection (after determination of lethal 50 dose, LD50) as following:

· Subgroup 1 (G1): Injected with 0.3ml (500mg/kg)

· Subgroup 2 (G2): Injected with 0.3ml (250mg/kg)

· Subgroup 3 (G3): Injected with 0.3ml (125mg/kg)

· Subgroup 4 (G4): Injected with 0.3ml (62.52mg/kg)

· Subgroup 5 (G5): Injected with 0.3ml (31.24mg/kg)

After 24 hours of injection, all mice were observed behaviorally and morphologically, and notes were recorded. Then, 48 hours after injection, the mice were euthanized; livers and kidneys were carefully removed and fixed in 10% buffered formalin solution. Next, the fixed biopsies were embedded in paraffin and cut into 5µm slices. The slices were mounted on glass slides and stained with hematoxylin and eosin for histological analysis. The images were examined under light microscope.

RESULTS:

Kinetic activity:

Observing animal behavior after injection of Sudafed showed more active locomotion and aggressiveness as well as loss of appetite especially at high conc.500mg/Kg.

Histology of livers and kidneys:

Section of kidney obtained from control animal showed normal histological structure of glomerulus (G), proximal convoluted tubules (P), and distal convoluted tubules (D) (Figure 1).

Figure 1: Section of kidney in control animal showing normal histology (H and E stain 400X).

In a section of normal liver, the parenchymal cells of liver (hepatocytes) are joined to one another in plates, with borders that face either the sinusoids or adjacent hepatocytes. Hepatocytes make contact with blood sinusoids, which are distensible vascular channels lined with highly fenestrated endothelial cells and populated with phagocytic Kupffer cells (Figure 2).

Figure 2: Section in the liver from control mice shows normal structure (H and E stain 400X).

Histological changes of liver in acute subgroups: Using 500mg/Kg Sudafed intraperitoneally, Liver sections show mild ballooning degeneration of hepatocytes and central vein congestion. Lower doses (250mg/Kg–31.42mg/Kg) revealed less prominent effect or no significant pathological changes (Fig3A-F).

Histological changes of kidney in acute subgroups:

With the 500mg/Kg Sudafed intraperitoneally, sections from the kidney show only mild hydropic swelling of tubular epithelium with congestion of intertubular blood vessels and relatively healthy glomeruli. Lower doses revealed no significant pathological changes (Fig 4 A-F).

DISCUSSION:

The present study demonstrated that intraperitoneal PSE increased kinetic activity and aggressive behavior as well as loss of appetite in addition to mild pathological changes in liver and kidney of mice. It is evident that there is a correlation between the dose administered and its behavioral effects¹⁰. Our results revealed that PSE is potent enough to act as a stimulant. Movement behavior in animals has been shown to increase following treatment with standard drugs, such as Kinetics¹¹, amphetamine, cocaine, modafinil¹², ethanol¹³, and morphine¹⁴. Enhancement of locomotion appears to be one of the most common effects of stimulant drugs. PSE, at a high dose (40mg/kg), was once reported to significantly replicate amphetamine effects in rats, according to drug discrimination analysis¹². Perfectly, drug discrimination analysis is used to determine whether a CNS chemical agent alters brain functions, leading to changes in mood, feelings, perceptions, and/or behaviors similar to those induced by a standard psychoactive drug, or results in neutral effects, similar to saline. As a sympathomimetic, PSE could potentially induce internal changes different from those induced by saline. In particular, PSE increases systolic blood pressure¹⁵and causes other neurological effects⁴, which may be the properties that allowed PSE to mimic amphetamine activity. Increased activity promotes resilience against compulsive cocaine seeking behavior¹⁶. These findings also suggest that the locomotor activity or reinforcement induced by stimulant drugs may be health risks associated with substances of abuse. A major concern associated with PSE use is illegal drug production. Several chemical methods have been developed to extract and convert PSE into amphetamine^17,18. Moreover, additional concerns have been raised based on research findings showing that PSE acts as a sympathomimetic. At the level of gene expression, PSE has been shown to exhibit stimulatory effects in the nucleus acumens and striatum, two major brain regions that are generally sensitive to drugs of addiction. These studies demonstrated the CNS effects of PSE injections, indicating that PSE effectively crosses the blood‑brain barrier¹⁹.

However, the present study showed mild changes in liver and kidney tissues especially at high concentration (500mg/Kg) of Sudafed. The findings after 24 hrs. of one intraperitoneal injection showed swelling, degeneration and congestion of both organs which boosts the opinion of toxic effect of PSE on body organs. It is worth to be mentioned that there are no previous works deal with the effect of this drug on liver or kidney.

THE MAJOR DRAWBACK OF THIS STUDY:

We were unable to make a firm conclusion with respect to the effect of pseudoephedrine on kinetic activity, and histology of Livers and Kidneys of albino mice due to the limitations of the published articles in this field.

CONCLUSION:

The present study demonstrated an evidence of pathological effects caused by high doses of PSE.A further study with larger sample sizes is required to determine the relationship between different doses of PSE and the systemic chronic effects.

ETHICAL CONSENT:

Ethical approval for this study was obtained from ethics committee of (College of Science, Dept. of Biology).

CONFLICT OF INTEREST:

There are no conflicts of interest.

ACKNOWLEDGMENT:

The authors of the article would like to thank everyone who provided assistance in accomplishing this work.

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Received on 09.05.2020 Modified on 12.11.2020

Research J. Pharm. and Tech. 2021; 14(9):5015-5018.

DOI: 10.52711/0974-360X.2021.00874