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ORIGINAL ARTICLE
Year : 2013  |  Volume : 3  |  Issue : 3  |  Page : 249-253

Momordica charantia (bitter melon) decreases serum/tissue lipid parameters in hyperammonemic rats


Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalainagar, Tamil Nadu, India

Date of Submission22-Oct-2012
Date of Acceptance20-Dec-2012
Date of Web Publication10-Jul-2013

Correspondence Address:
P Subramanian
Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalainagar - 608 002, Tamil Nadu
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2231-0738.114843

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   Abstract 

Introduction: Ammonia is present in all living organisms as a byproduct of the degradation of proteins and other nitrogenous compounds. At high levels ammonia is toxic, causing functional disturbances in the central nervous system that could even lead to coma and death. The present study evaluates the efficacy of an ethanolic extract of Momordica charantia L. (MCE) fruits in decreasing serum and tissue lipids and body weight in rats with experimentally-induced hyperammonemia. Materials and Methods: Experimental hyperammonemia was induced in adult male Wistar rats (180-200 g) by intraperitoneal (i.p.) injections of ammonium chloride (100 mg/kg body weight) thrice a week. The effect of oral administration (thrice a week for eight consecutive weeks) of MCE (300 mg/kg body weight) on lipid profiles in serum and tissues (brain and liver) of normal and experimental animals were analyzed. Results and Discussion: The levels of serum and tissue cholesterol, triglycerides, free fatty acids, and phospholipids were significantly increased in ammonium chloride - induced hyperammonemic rats. The administration of MCE to ammonium chloride - treated rats significantly restored all these changes to almost normal levels. Ammonium chloride - treated rats showed increases in body weight compared to the control rats. The exact mechanism of this antihyperlipidemic effect of MCE needs to be investigated and the active constituents identified and isolated.

Keywords: Ammonium chloride, hyperammonemia, lipid profile, Momordica charantia, Wistar rats


How to cite this article:
Thenmozhi A J, Subramanian P. Momordica charantia (bitter melon) decreases serum/tissue lipid parameters in hyperammonemic rats. Int J Nutr Pharmacol Neurol Dis 2013;3:249-53

How to cite this URL:
Thenmozhi A J, Subramanian P. Momordica charantia (bitter melon) decreases serum/tissue lipid parameters in hyperammonemic rats. Int J Nutr Pharmacol Neurol Dis [serial online] 2013 [cited 2019 Nov 15];3:249-53. Available from: http://www.ijnpnd.com/text.asp?2013/3/3/249/114843


   Introduction Top


Ammonia is an important source of nitrogen and is required for amino acid synthesis. It is also necessary for maintenance of normal acid - base balance. When present in high concentrations, ammonia is toxic. Ammonia is a neurotoxin that accumulates in the brain in a variety of neurological disorders associated with hyperammonemia. [1],[2] Endogenous ammonia intoxication can occur when there is impaired capacity of the body to excrete nitrogenous waste, as seen in congenital enzymatic deficiencies. Patients with urea cycle defects (UCD), organic acidemias, fatty acid oxidation defects, bypass of the major site of detoxification (liver), and Reye syndrome, as well as patients in the post-chemotherapy state or those who have suffered exposure to various toxins and drugs can all present with elevations in ammonia levels. [3],[4] Hyperammonemia refers to a clinical condition characterized by elevated serum ammonia levels. It manifests with hypotonia, seizures, emesis, and abnormal neurologic changes. [5],[6] Hyperammonemia can cause irreparable damage to the developing brain. The presenting symptoms include posturing, cognitive impairment, seizures, and cerebral palsy. Delay in diagnosis and treatment of hyperammonemia, irrespective of the etiology, can lead to neurologic damage and even death, and thus it should be considered a medical emergency when it presents. [5],[7],[8]

Valporic acid, phenobarbitol, and carbamazepine are some of the currently used antiseizure and antihyperammonemic drugs. These drugs or therapies are sometimes associated with inconvenient side effects, high cost, and inadequate efficacy. [9] Therefore the search for newer and better drugs with antihyperammonemic activity is continuing, and the traditional medicinal plants offer rich possibilities.

Momordica charantia L. (bitter melon) is one of the most important species of the family Cucurbitaceae. Its fruit as well as the other parts of the plant have medicinal value. [10],[11] The fruit is consumed as part of the diet but they are also reported to possess a wide range of pharmacological activities; for example, hypoglycemic, [12] antidiabetic, [13] antifungal, [14] ability to inhibit p-glycoproteins, [15] antihyperlipidemic, [16] and antioxidant effects have all been reported. [17] The fruit has been used traditionally as an anthelmintic, antiemetic, carminative, and purgative, as well as for the treatment of anemia, jaundice, malaria, cholera, etc. [18] Examination of the phytochemicals of this plant indicates the presence of various active components: momorcharins, momordenol, momordicilin, momordicins, momordicinin, momordin, momordolol, charantin, charine, cryptoxanthin, cucurbitins, cucurbitacins, cucurbitanes, cycloartenols, diosgenin, elaeostearic acids, erythrodiol, galacturonic acids, gentisic acid, goyaglycosides, goyasaponins, and multiflorenol, have all been isolated from the plant. [19],[20] It is a well-documented fact that most medicinal plants are rich in phenolic compounds and bioflavonoids with excellent antioxidant properties. [21],[22]

Multiple studies have reported that bitter melon is capable of suppressing weight gain and reducing adiposity in normal animal models fed with a High Fat diet for a short period. [23],[24] The present study evaluates the effect of an ethanolic extract of M. charantia fruits on experimentally induced hyperammonemia in rats to determine its effect on body weight changes and serum and tissue (brain and liver) lipid levels (i.e., cholesterol, triglycerides, free fatty acids, and phospholipids).


   Materials and Methods Top


Plant material

Mature green M. charantia were collected from Chidambaram, Cuddalore District, Tamil Nadu, India. The plant was identified and authenticated at the Herbarium of the Botany Directorate in Annamalai University. A voucher specimen (No. 1260) was deposited at the Botany Department of Annamalai University.

Preparation of the alcoholic extract (MCE)

Alcoholic extract of the fruit was prepared according to the method developed by Shibib et al. [25] One kilogram of unripe fruit bought from the local market was thoroughly washed and the seeds were removed. The pulp was blended in 1500 ml of 95% alcohol and left at room temperature, with occasional shaking, for 48 h. The suspension was filtered through cheesecloth, and the filtrate was evaporated in a Rotovac™ (BÜCHI Labortechnik AG, Switzerland) at 40°C to remove the alcohol. The final residue was stored at −20°C. It was suspended in 1% (w/v) carboxymethyl cellulose (CMC) for use in the investigation. [26]

Chemicals

Ammonium chloride was purchased from Sisco Research Laboratories, Mumbai, India. All the other chemicals used in the study were of analytical grade.

Animals

Adult male albino Wistar rats, weighing 180-200 g, bred in the Central Animal House, Rajah Muthiah Medical College, Annamalai University, were used for the experiment. The animals were housed in polycarbonate cages in a room with a 12-hour day-night cycle, at a temperature of 22 ± 2°C and humidity of 45-64%. The animals were fed a standard pellet diet (Hindustan Lever Ltd., Mumbai, India) and water ad libitum. The animal experiment was approved by the Ethical Committee of Annamalai University (clearance No. 536/20-03-08), and all procedures were in accordance with the guidelines of the National Institute of Nutrition (NIN), Indian Council of Medical Research (ICMR), Hyderabad, India. Hyperammonemia was induced in the rats by intraperitoneal injections of ammonium chloride (100 mg/kg body weight) thrice a week for 8 consecutive weeks. [27]

Experimental design

A total of 32 rats were used in the experiment. The rats were divided into four groups of eight rats each. Group I rats received 1% (w/v) CMC and were considered as the control group; group II rats were administered with MCE (300 mg/kg body weight) using an intragastric tube; [25] group III rats were treated with ammonium chloride intraperitoneally (100 mg/kg body weight); [27] and group IV rats were treated with ammonium chloride (100 mg/kg) and also administered MCE (300 mg/kg). At the end of eighth week, the rats were fasted overnight and killed by cervical dislocation after anesthetizing with intraperitoneal ketamine hydrochloride (30 mg/kg body weight). Blood was collected and plasma and serum were separated by centrifugation. The liver and brain tissues were excised immediately and rinsed in ice-chilled normal saline. About 500 mg of the tissues were homogenized in 5.0 ml of 0.1 M Tris-HCl buffer (pH 7.4). The homogenate was centrifuged and the supernatant was used for the estimation of various biochemical parameters.


   Results Top


Ammonium chloride-treated rats showed increases in body weights compared to the control rats. Oral treatment with MCE in ammonium chloride-treated rats caused significant decrease in the weight gain as compared to rats treated with ammonium chloride alone [Table 1]. The levels of serum cholesterol, triglycerides, free fatty acids, and phospholipids were significantly increased in the ammonium chloride-induced hyperammonemic rats, and the administration of MCE significantly restored all these changes to almost normal levels [Table 2]. [Table 3] and [Table 4] shows the effect of MCE on the changes in the levels of tissue cholesterol, triglycerides, free fatty acids, and phospholipids in normal and experimental rats. Ammonium chloride - induced rats showed significantly increased levels of these biochemical parameters when compared with normal rats. Oral administration of MCE to ammonium chloride - treated rats significantly decreased the levels of tissue lipids.
Table 1: Body weight changes in each group

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Table 2: Changes in the levels of serum lipid profi le in control and treated rats

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Table 3: Changes in the levels of tissue cholesterol and triglycerides in control and treated rats

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Table 4: Changes in the levels of tissue free fatty acids and phospholipids in control and treated rats

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   Discussion Top


Our results showed that rats treated with ammonium chloride gained significantly more body weight than did control rats. The observed weight gain might be attributed to the increased levels of lipid profile, aminoacids and proteins during hyperammonemic conditions. [28],[29],[30] Group 4 (MCE + ammonium chloride) rats showed body weight similar to that of group 1 control rats, which might be due to the antilipidemic effects of MCE. [31],[32] Numerous animal studies have indicated the efficacy of bitter melon in the amelioration of weight gain and regulation of lipid metabolism. [23],[33]

Ammonia is a normal byproduct of the degradation of proteins and other compounds; however, at high concentrations, ammonia is toxic and leads to functional disturbances of the central nervous system. The toxic effects of ammonia are not seen in the healthy as it is detoxified in the liver by incorporation into urea for elimination in urine. [34],[35] Increased levels of circulatory ammonia and urea indicate a hyperammonemic condition in ammonium chloride-induced rats, [36] which may be due to liver damage caused by ammonia intoxication. Numerous investigations have documented that plant extracts containing phenolic compounds and flavonoids can remove excess ammonia, urea, uric acid, and creatinine during disease conditions such as hyperammonemia, nephrotoxicity, etc. [37],[38] Administration of MCE to ammonium chloride-induced hyperammonemic rats significantly decreased the levels of blood ammonia and urea [32] (data not shown). The reduction in the levels of ammonia and urea during MCE treatment shows the potent anti-hyperammonemic effect of MCE. [32]

In the present study, elevated levels of ammonia caused a significant rise in serum and tissue lipids (cholesterol, triglycerides, phospholipids, and free fatty acids). These findings indicate that hyperammonemia is accompanied by dyslipidemia. Previous studies from our lab have also demonstrated the elevation of levels of serum and tissue lipids during hyperammonemic conditions. [29],[39] It was reported that ammonium (chloride/acetate) salts may deplete levels of α-KG and other Krebs cycle intermediates [29],[40] and thus elevate the levels of acetyl coenzyme A. The elevated levels of acetyl coenzyme A may increase the levels of lipids (free fatty acids, triacylglycerols, phospholipids, and cholesterol) as observed in our study. Another important function of α-KG is in the formation of carnitine. [34],[40] Previous findings indicate that carnitine deficiency results in hyperammonemia. [41],[42] Carnitine acts as a carrier of fatty acids into cell mitochondria so that proper catabolism of fats can be carried out. [29],[35] The decreased levels of α-KG and other Krebs cycle intermediates in rats treated with ammonium chloride might have led to the accumulation of fatty acids. [29]

Lowering of serum/tissue lipid levels through dietary measures or drug therapy seems to be associated with a decrease in the risk of various vascular diseases. [43],[44] In the current study, MCE treatment of hyperammonemic rats caused a significant decrease in serum and tissue lipids (cholesterol, triglycerides, phospholipids, and free fatty acids). The effect of MCE on controlled mobilization of serum cholesterol, triglycerides, phospholipids, and free fatty acids was presumably mediated through the regulation of the hydrolysis of certain lipoproteins and their selective uptake and metabolism by different tissues.

Phenolic compounds have the ability to normalize the levels of serum and tissue lipids during disease conditions. [45] Quercetin, a polyphenol found in the fruits of M. charantia, has been shown to significantly lower triacylglycerols and free fatty acids in obese rats. [46] It was reported that M. charantia could elevate the activities of hepatic and muscle mitochondrial carnitine palmitoyl transferase-I (CPT-I) and that it attenuates fatty acyl carnitine accumulation in muscle, increasing the ability of muscles to oxidize fatty acids. [23] These findings support the results of our experiments.


   Conclusion Top


The elevations in serum and tissue lipids during experimental hyperammonemia were restored to near normal levels by MCE treatment. M. charantia has been shown to possess both hepatoprotective and hypolipidemic effects in rats with alloxan-induced diabetes. [30],[32] It is possible that M. charantia could modulate lipid levels in the hyperammonemic condition and enhance proper metabolism of fats.

 
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    Tables

  [Table 1], [Table 2], [Table 3], [Table 4]


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