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8.
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GENERAL DISCUSSION:
7.1 Need for the study :
The liver is a vital organ. Liver intoxication has increased as a result of exposure to high levels of environmental toxins, as the liver has an important role in detoxification. Death rate extrapolations for USA for Liver failure: 60,000 per year, 5,000 per month, 1,153 per week, 164 per day, 6 per hour. Viral hepatitis causes 95- 100% of Acute hepatic failure in India. Liver damage is always associated with cellular necrosis, increase in tissue lipid peroxidation and depletion in the tissue GSH levels. In addition serum levels of many biochemical markers like Aspartate aminotransferase (AST), Alanine-aminotransferase (ALT), triglycerides, cholesterol, bilirubin, Alkaline phosphatase, are elevated.1, 2
The manifestations of drug-induced hepatotoxicity are highly variable, ranging from asymptomatic elevation of liver enzymes to fulminant hepatic failure. Paracetamol (PCM) also known as Acetaminophen, taken in overdose can cause severe hepatotoxicity and nephrotoxicity. PCM is activated and converted by cytochrome P450 enzymes to toxic metabolite NAPQI (N-acetyl-p-benzoquinoneimine) that causes oxidative stress and glutathione (GSH) depletion. In spite of tremendous advances in modern medicine, there are hardly any reliable drugs that protect the liver from damage and/or help in regeneration of hepatic cell. Many active plant extracts are frequently utilized to treat a wide variety of clinical diseases including liver disease.3 Therefore, searching for effective and safe drugs for liver disorders are continues to be an area of interest.
Ethanol produced significant changes in physical (increased liver weight and volume), biochemical (increase in serum alanine transaminase, aspartate transaminase, alkaline phosphatase, direct bilirubin, total bilirubin and triglycerides), liver tissue (decreased total protein, GHS and increased MDA), histological (hepatocytes damage) and functional (thiopentone induced sleeping time) liver parameters.4,5
Paradoxically, these may themselves cause hepatic damage such as cholestatic jaundice with azathioprine and elevation of serum transaminase by interferon and virazole.6 It is therefore imperative to search alternative drugs for treatment of hepatic diseases to replace the currently available drugs with suspicious efficacy and safety.
Herbs have recently attracted attention as health beneficial foods and as source materials for drug development. Herbal medicines derived from plant extracts are being increasingly utilized to treat a wide variety of clinical diseases including liver disease, ischemia, reperfusion injury, atherosclerosis, acute hypertension, hemorrhagic shock, diabetes mellitus and cancer with relatively little knowledge regarding their modes of action.
Mesua ferrea plant belongs to family Clusiaceae which mainly contains mesuferrone-A and B, mesuaferrol, mesuanic acid, a- and ß-amyrin, and ß-sitosterol. The seeds and heartwood contain a number of xanthones and coumarins. Methanolic extract of Mesua ferra linn evaluated for their antioxidant and hepatoprotective. Coumarins, mainly responsible for hepatoprotectivity.
Coumarins structure
Mesua ferrea is medium sized to large evergreen tree with very hard, strong and long wood. It is common in wet zone of Srilanka, Assam, southern China and peninsula. Mesua ferrea shows anti-inflammatory and antiseptic activity. Almost every part of the tree has therapeutic value in one or another disorder. The flowers are useful in medical conditions like asthma, leprosy, cough, fever and impotency. Dried flowers are useful in bleeding haemorrhoids and dysentery with mucus. The volatile oil extracted from plant can be used for sores, scabies, wounds, and rheumatism. It grows in the Himalayas from Nepal eastwards, in north-eastern India, the Deccan Peninsula and the Andaman Islands.7
The leaves are applied to the head in the form of a poultice for severe colds. Oil from the seeds is used for sores, scabies, wounds, and rheumatism. The root of this herb is often used as an antidote for snake poison. Phenolic extract of seed oil of M. ferrea revealed potent antiasthmatic effect.
There are so many allopathic drugs available for hepatoprotective, but along with therapeutic uses there will be the adverse effects also. so, Herbal drugs will be the very much useful. Up to now there is no hepatic activity done on this plant isolated Mesuol product regarding hepatoprotectivty.This is the main reason why i have selected this plant to my activity.
8.1 Review of literature
Botanical Name: Mesua ferrea
Kingdom : Plantae
Division : Magnoliophyta
Class : Magnoliopsida
Order : Malpighiales
Family : Clusiaceae
Subfamily : Kielmeyeroideae
Tribe : Calophylleae
Genus : Mesua
Species : M. ferrea.8
Synonyms
Cobra's saffron, nagakesara, nagkeshar cobra saffron, nagachempakam, nagakesara, nangu.
Parts Used
Flower buds, flowers, fruit, seed, root, bark, oil.
Distribution
It grows in the Himalayas from Nepal eastwards, in north-eastern India, the Deccan Peninsula and the Andaman Islands, ascending to an altitude of 1,500 m.A medium-sized to large evergreen tree with short trunk, often buttressed at the base. The bark is greyish or reddish brown; the leaves are lanceolate, coriaceous, generally covered with
a waxy bloom underneath, red when young; the flowers are large, white and fragrant; the fruits are ovoid, nearly woody; the seeds are dark brown.
Product Range
Lukol, Pilex (VeinCare), Styplon, Styplon Vet, Chyavanaprasha
Chemical Constituents
The stamens which yield the drug nagakesara contain mesuferrone-A and B, mesuaferrol, mesuanic acid, a- and ß-amyrin, and ß-sitosterol. The seeds and heartwood contain a number of xanthones and coumarins. Volatile oil from the flowers showed antibacterial, antifungal and anthelmintic activities. The Methanolic extract showed antioxidant and hepatoprotective activity.9
Ethnomedical uses
The leaves are applied to the head in the form of a poultice for severe colds. Oil from the seeds is used for sores, scabies, wounds, and rheumatism. The root of this herb is often used as an antidote for snake poison. The dried flowers are used for bleeding haemorrhoids and dysentery with mucus. Phenolic extract of seed oil of M. ferrea revealed potent antiasthmatic effect. Volatile oil from the flowers showed antibacterial, antifungal and anthelmintic activities.
Fresh flowers are also prescribed for excessive thirst, excessive perspiration, cough, and for indigestion.10 A number of compounds, representing a variety of chemical structures isolated from the carious parts of this plant, may provide interesting leads for further pharmacological evaluation. Flowers for Antioxidant, hepatoprotectivity.11
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Objective of the studies are
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To Extraction and Isolation of Mesuol.
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To investigate the effect of Mesuol on paracetamol induced hepatotoxicity by measuring the following Parameters.
Aspartate aminotransferase (AST),
Alanine-aminotransferase (ALT),
Alkaline-phosphatase (ALP),
Bilirubin,
Lipid-peroxidation, Superoxide dismutase,
Glutathione,
Histopathological Examination.
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To investigate the effect of Mesuol on ethanol induced hepatotoxicity by measuring the following Parameters.
Aspartate aminotransferase (AST),
Alanine-aminotransferase (ALT),
Alkaline-phosphatase (ALP),
Bilirubin,
Lipid-peroxidation, Superoxide dismutase,
Glutathione,
Histopathological Examination.
MATERIALS AND METHODS:
9.1 Extraction and Isolation of Mesuol:
The oil obtained by petroleum ether extract of the seed kernel by using Soxhlet apparatus. The expressed oil from the seed kernels was kept in a refrigerator for three weeks when crud Mesuol was deposited. On repeat crystallization it formed pale yellow rectangular plates of m.p.1540. The compound was found homogeneous when examined paper chromatographically.12
METHODS:-
9.2 Paracetamol induced Hepatotoxicity.
Male Wistar rats weighing 150-180 g are used for the experiment. The animals are housed under conditions of controlled temperature and 12 hour day-night cycle and are fed standard chow.
Thirty six animals were randomly divided equally into six groups of six animals each.
Group 1: (Control) receives normal saline orally (0.2 ml/100 g) for 16 consecutive days.
Group 2: (Drug control) receives Mesuol.
Group 3: (PCM-induced control) receives paracetamol (750 mg/kg p.o.)
at every 72 h for 10 days.3
Group 4: (Pre-treatment experiment)receives Mesuol and treated with paracetamol on Days 14, 15, and 16 of the treatment period.
Group 5: (Post-treatment experiment) receives Mesuol and treated
with paracetamol on Days 1, 2, and 3 of the treatment period.
Group 6: (Silymarin control) receives Silymarin (100 mg/kg bw) for 16 consecutive days and treated with paracetamol on Days 1, 2, and 3 of the treatment period.
Twenty-four hours after the last treatment the rats were euthanized.
Blood was collected by means of a heart puncture on the end of the treatment in the pre- or post-treated groups, respectively. Plasma was separated by centrifugation at 900 rpm for 10 minutes at 4°C, and used for measuring the Aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatise (ALP), and bilirubin concentration using the auto analyzer. Lipid peroxidation, superoxide dismutase, glutathione are assayed using tissue and Histopathological changes will be carried out.
9.3 Ethanol induced hepatotoxicity.
Male Wistar rats weighing 150-180g are used for the experiment. To evaluate the hepatoprotective effect of Mesuol. Rats were administered either saline or Mesuol or Silymarin(100 mg/kg bw) for 3 days and subsequently exposed to the treatment of ethanol after the last Mesuol/saline/silymarin treatment.4,5
Thirty six animals were randomly divided equally into six groups of six animals each.
Group 1: (Control) receives normal saline orally (30 ml/kg/day) for 20 days.
Group 2: (Drug control) receives Mesuol
Group 3: (Ethanol treated) receives ethanol (36.6 % v/v) 30 ml/kg/day and corn oil (10 ml/kg/day) in three divided doses for 20 days.
Group 4: (Low dose) receives low dose of Mesuol before treating with ethanol
(36.6 % v/v) 30 ml/kg/day and corn oil (10 ml/kg/day) in three divided doses.
Group 5: (High dose) receives high dose of Mesuol before treating with ethanol
(36.6 % v/v) 30 ml/kg/day and corn oil (10 ml/kg/day) in three divided doses.
Group 6: (Silymarin control) receives Silymarin (100 mg/kg bw) for 3 consecutive days before treating with ethanol on Days 1, 2, and 3 of the treatment period.
After 24 hrs of treatment the blood collection done by means of a heart puncture and serum was separated by centrifugation. In serum, Alanine aminotransferase (ALT), Aspartate aminotransferase (AST) and Lactate dehydrogenase (LDH) and in tissue, Lipid peroxidation, SOD and glutathione were assayed and Histopathological studies will be carried out.
Statistical Analysis
The analysis of data will be done by using suitable statistical methods.
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