EVALUATION OF ANTIDOTE ACTIVITY OF LEAVES OF MUSA SAPIENTUM LINN. ON RATS

 

About Authors:
Vishnu Dev 1*, Rajiv saxena 2
1 Truba Institute of pharmacy, Bhopal.
2 Assoc. Professor, Truba Institute of pharmacy, Bhopal
*vishnudev2007@rediffmail.com

Abstract
The present study was designed to investigate the anti-dote activity of the methanolic and aqueous extract of musa sapientum. The extract was evaluated by physical and serum biochemical parameters as TC and TG. Anti-dote activity of both extract was determined by acute and chronic codeine poisoning,
Acute opioid intoxication and overdose are common causes of presentation to emergency departments. Although naloxone, a pure opioid antagonist, has been available for many years, there is still confusion over the appropriate dose and route of administration. This article looks at the reasons for this uncertainty and undertakes a literature review from which a treatment algorithm is presented.
The anti-dote activity of musa sapientum leaves against codeine induced acute and chronic poisoning was studied in Wistar albino rats. The various fractions of the leaf extract were administered orally (200 mg/kg) simultaneously with codeine (32m g/kg) for 5-10 days. At the end of the experiment, levels of triglycerides, total cholesterol content were measured. Administration of codeine produced a CNS depressant activity and causes poisioning. There was a significant increase in total cholesterol and triglycerides when compared to normal control. physical observation indicated that simultaneous administration of the fractions delayed the onset of toxicity. All the fractions prevented the poisoning damage caused codeine overdose. The leaves of musa sapientum protected the CNS against codeine damage which may be due to its antagonist activity of codeine.

REFERENCE ID: PHARMATUTOR-ART-1656

1. Introduction-
Opiate poisoning can occur at any time from birth (when pethidine given to the mother in labour may suppress ventilation) to terminal care. The outcome can range from discomfort like constipation to death from respiratory depression.(Paul harris et al,2001).

Codeine or 3-methylmorphine (a natural isomer of methylated morphine, the other being the semi-synthetic 6-methylmorphine) is an opiate used for its analgesic, antitussive and antidiarrheal properties. Codeine is the second - most predominant alkaloid in opium, at up to three percent; it is much more prevalent in the Iranian poppy (Papaver bractreatum), and codeine is extracted from this species in some places although the below-mentioned morphine methylation process is still much more common. It is considered the prototype of the weak to midrange opioids (tramadol, dextropropoxyphene, dihydrocodeine, hydrocodone). (Vale JA. et al, 2004)

Opium derivatives are CNS depressants that probably act on the sensory cortex or higher centers and thalami. Because they can relieve pain, change or elevate mood, relieve tension, fear, and anxiety and produce feelings of peace euphoria, and tranquillity, they are particularly likely to lead to physical and psychologic dependence.(Paul Harris et al, 2001)

Musa sapientum leaf belongs to the Musaceae  family is the largest herbaceous flowering plant. The plants are normally tall and fairly sturdy and are often mistaken for trees, but their main or upright stem is actually a pseudostem that grows 6 to 7.6 meters (20 to 24.9 ft) tall, growing from a corm Banana leaf is the leaf of the Banana plant. It is used as a decorative element for special ceremonies in Hindu and Buddhist cultures. It is also used as a plate to serve food in countries like India. Banana leaves, though commonly thrown away, contain large amounts of polyphenols, including EGCG, similar to green tea. Traditionally the stem of banana and leaf was used as treatment in opium poisoning. The leaf are also used in the fish pool to avoid from poisoning. (Grollman AP, et al, 2007).

2. Materials and Methods
Experimental animals: Wistar
albino rats of either sex weighing between 125-150 g were used. They were housed in polypropylene cages inside a well-ventilated room. The room temperature was maintained at 23 ± 2° C with a 12 hours light/dark cycle. Food and water were given ad libitum.

2.1 Drugs and chemicals: codeine was obtained from the local market of Bhopal which was zydus alidac company with 15 mg/tab of dose. Naloxone was obtained from the triokaa pharmaceutical ltd. Banglaore.

2.2 Plant material: The plant material consists of dried powdered leaves of musa sapientum belonging to the family musaceae. The leaves of musa sapientum were collected from bhopal district, m.p., India during the month of November 2011. The plant was identified and authenticated by Mr. Zia Ul Hasan, botanist safia college of science, Bhopal, Madhya Pradesh , bearing the reference number 345/Bot/safia/2012.

2.3 Preparation of extract and fractionation: Fresh leaves of the plant were dried in shade under room temperature, powdered mechanically and sieved through No. 20 mesh sieve and extracted with methanol and water. Powder was packed in filter paper wrapped inside soxhlet tube and introduced in the extraction unit of Soxhlet extractor ,extraction was done with ethanol(80 ml) and water(80 ml) for 25 cycles. The extracts were filtered and concentrated with rotary evaporator and residual fractions (RF) of musa sapientum were used for the study.(khandelwal et al, 2008).

2.4 Acute toxicity studies: Swiss albino mice weighing between 20-30 g maintained under standard laboratory conditions was used. Animals were divided into four groups consisting of 5 each; the animals received a single oral dose (2,000 mg/kg, body weight) of each fraction. Animals were kept overnight fasting prior to drug administration. After the administration of the fraction, food was withheld for further 3-4 hours. Animals were observed individually at least once during the first 30 min after dosing, periodically during the first 24 hours (with special attention during the first 4 hours) and daily thereafter for a period of 14 days.(OECD guideline 423)

2.5 Selection of dose of the extract: LD50 was done as per OECD guidelines for fixing the dose for biological evaluation (OECD, 2000). The LD50 of the fractions as per OECD guidelines falls under category 4 values with no signs of acute toxicity at doses of 2,000 mg/kg. The biological evaluation of the fractions was carried out at a dose of 200 mg/kg body weight.

Experimental protocol: Animals were divided into 6 groups consisting of six animals each. Group I received saline water and served as control. Group II received codeine (32 mg/kg orally) alone (negative control). Groups III received the nalaxone (2 mg/kg) with codeine(positive . group IV and V recieved musa sapientum  respectively at a dose of 200 mg/kg orally simultaneously with codeine. All the drugs were administered for a period of 5 day for acute and 10 days for chronic group.

2.6 Examination procedure for total cholesterol-

Pipette in to tube marked

blank

Standard

test

Serum/plasma

...........

.........

10 µl

Reagent 2

..........

10 µl

..........

Reagent 1

1000 µl

1000 µl

1000 µl

Mixed well incubate at 370Cfor 10 minutes

Analyzer was programmed as per assay parameters.

  1. Blank the analyzer with reagent blank.

  2. Measured absorbance of standard followed by the test.

  3. Calculate the result as per given calculation formula.

                                                    Absorbance of test
Cholesterol concentration (mg/dl) =  ---------------------- × 200
                                                 Absorbance of standard  

2.6 Examination procedure for triglycerides

Preparation of working reagent:

The content of enzyme reagent was dissolved in 10 ml of diluents buffer. The working reagent    was stabilize for 4-6 week at 2-80 C.

Pipette in to tube marked

blank

Standard

test

Serum/plasma

...........

.........

10 µl

Reagent 2

..........

10 µl

..........

Reagent 1

1000 µl

1000 µl

1000 µl

2.7 Collection of blood samples
On 5th day of treatment of acute group and 10th day of chronic group treatment, the blood was collected by retro orbital eye plexuses puncture, under mild chloroform anaesthesia in tubes. Serum was obtained by centrifugation of blood samples within one hours of collection using  ultra centrifuge (Remi) at 2000 rpm for 30 minutes between 4-80C temperature and directly used for estimating serum lipid profile ( TC , TG).

Statistical analysis: Statistical analysis was carried out by one-way analysis of variance (ANOVA) followed by Dunnett’s test. Results are expressed as mean ± SEM from six rats in each group. P values <0.05 were considered significant.

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