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Summary An Improvement in Hepatoprotective Activity by Herbal Drug Combination

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MATERIALS AND METHODS Drugs and Chemicals All Reagents Procured Were Analytical Grade: Paracetamol tablet (Sun Pharmaceuticals Ltd) purchased from a drugstore. Total Bilirubin, Total Proteins, SGOT, SGPT, Alkaline Phosphate were assayed by using kits from Ranbaxy diagnostic, New Delhi. Plant Collection: Fresh leaves of cucumerina L. and sativum fruits were collected from Komarapalayam and authenticated by Dr.P. Satyanarayana, Scientist D & Head office in charge, Southern Regional Centre, TNAU campus, Coimbatore. The leaves were cut into small pieces and air dried indoor subdued light and with good ventilation and then crushed into a fine powder using a laboratory Homogenizer, which passed through 22 No sieve. The fruits of sativum were dried and then crushed into fine powder by using laboratory homogenizer then stored for further use. Preparation of Plant Extracts Ethanol Extract of Cucumerina Linn. (EETC): Fine powdered Leaves of cucumerina were extracted successively with petroleum ether and Ethanol (60-80°C) using Soxhlet apparatus. The extract was filtered and evaporated to separate solvent and residue. The semisolid residue thus obtained was stored in desiccator until further use. Ethanol Extract of Sativum: (EECS): Fine powdered fruits of sativum were extracted successively with petroleum ether and ethanol (60-80°C) using Soxhlet apparatus in access standard pellet diet and water ad libitum. Experimental Protocol Acute Oral Toxicity Studies [12, 13]: The acute oral toxicity study was followed by using OECD GUIDELINES - 423 (Organization of Economic Co-operation and Development) - Fixed dose procedure (FDP). Acute toxicity study was performed for ethanol extract of Cucumerina L. (EETC) and mixtures of ethanol extract of Cucumerina L. with ethanol extract of Sativum (MEETC&EECS) extract according to the acute toxic classic method as per OECD (423) guidelines5, albino rats were used for acute toxicity study. The animals were kept in fasting condition for overnight providing only water, then the extract was administered orally at the dose of 50, 100, 200 and 500 mg/kg and observed for 16 days. If death was observed in 2 out of 3 animals, then the dose administered was concluded as toxic dose. No death was occurring up to the dose level of 500mg/kg. Hepatoprotective Activity [14-21]: Group 1: Normal control, received 0.5% Carboxy methyl cellulose (CMC) solution (1ml/kg) once daily for 7 Days. Group 2: Hepatotoxient, administered with paracetamol (3gm/kg) a single dose on day 7. Group 3: Receives Trichosanthes cucumerina extract (150mg/kg) once daily for 7 days. (EETC). Group 4: Receives combination of ethanol extract of Trichosanthes cucumerina extract and coriandrum sativum extract in 1:1 ratio(150mg/kg) for 7 days. MEETC&EECS. Table 1: Effect of Extracts in Paracetamol induced hepatotoxicity rates Parameters Control Paracetamol EETC EETC&EECS Standard BILIRUBIN(d/l) 0.31±0.17 1.89±0.12*** 0.79±0.13*** 0.35±0.27*** 0.34±0.16*** SGOT (U/l) 135.03±12.21 419.65±25.93*** 218.83±14.12*** 195±14.75*** 171.06±17.75*** SGPT (U/l) 63.73±10.94 282.58±10.31*** 138.11±10.96*** 79.5±17.78*** 68.31±7.44*** ALP (U/l) 170.33±22.22 436.51±27.07*** 226.10±17.39*** 198.5±30.02*** 175.88±22.84*** *p 0.05; ** p 0.01; *** p 0.001. Values are expressed as mean (standard deviation; SD). Statistical significance was calculated with ANOVA followed by tukey compare all pair of coloum treated group with paracetamol group Group 5: Standard drug control, receives silymarin (100 mg/kg) once daily for 7 days (Std ). Group 3 to Group 4 receives paracetamol (3gm/kg ) as a single dose on day 3, Thirty minutes after administration of drug extract and silymarin respectively. Assessment of Hepatoprotective Activity: All the extracts and the paracetamol were administered orally by suspending in 0.5% CMC solution. Animals were

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An Improvement in Hepatoprotective Activity by
Herbal Drug Combination




INTRODUCTION caused by overdose of Paracetamol (39%) and idiosyncratic
liver injury triggered by other drugs
The liver is one of the major organs in human body (13%) [5]. Drugs play an vital role in cause of liver injury.
which plays an important role in intensifying metabolism Some surveys indicate that approximately 75% of the
and excretion [1, 2] and it has an amazing role in the idiosyncratic drug reaction results in liver transplantation
preservation, performance and regulating homeostasis or even death [6]. It also refer that the rate of
of the body. It is occupied almost all the biochemical hepatotoxicity reported that are much higher in
pathways of development, fight against all diseases, developing countries like India (8% - 30%) compared to
nutrient supply, reproduction [3] and energy provision. that in advanced countries (2% - 3%) with a similar dose
The liver is commonly not only to carry out schedule [7]. In spite of incredible advanced in
physiological functions, but also to shield against the modern medicine, there are hardly any reliable drugs
toxic effect of harmful drugs [4]. Due to wonderful that protect the liver from damage and help in
efficient development in the field of hepatology, liver regeneration of hepatic cell. Many active plant extracts are
problem is one of are on the rising problem in recent recurrently employing to treat a wide range of diseases
years. Drug-induced liver injury is a major health problem including liver disease. Herbal medicines are believed to
take place in health care professionals, pharmaceutical be much safer and proved elixir in the treatment on
industry and drug regulatory agencies. The United States various ailments [8]. Therefore, searching for effective
Acute Liver Failure Study Group, survey referred that 50% and safe drugs for liver disorders are continuing to be an
of acute liver failure take place by including hepatotoxicity area of interest.




1454

, access standard pellet diet and water ad libitum.

MATERIALS AND METHODS Experimental Protocol
Acute Oral Toxicity Studies [12, 13]: The acute oral
Drugs and Chemicals toxicity study was followed by using OECD GUIDELINES
All Reagents Procured Were Analytical Grade: - 423 (Organization of Economic Co-operation and
Paracetamol tablet (Sun Pharmaceuticals Ltd) purchased Development) - Fixed dose procedure (FDP).
from a drugstore. Total Bilirubin, Total Proteins, SGOT, Acute toxicity study was performed for ethanol extract
SGPT, Alkaline Phosphate were assayed by using kits from of Cucumerina L. (EETC) and mixtures of ethanol extract
Ranbaxy diagnostic, New Delhi. of Cucumerina L. with ethanol extract of Sativum
(MEETC&EECS) extract according to the acute toxic
Plant Collection: Fresh leaves of cucumerina L. and classic method as per OECD (423) guidelines5, albino rats
sativum fruits were collected from Komarapalayam and were used for acute toxicity study. The animals were kept
authenticated by Dr.P. Satyanarayana, Scientist D & Head in fasting condition for overnight providing only water, then
office in charge, Southern Regional Centre, TNAU campus, the extract was administered orally at the dose of 50, 100,
Coimbatore. The leaves were cut into small pieces and air 200 and 500 mg/kg and observed for 16 days. If death was
dried indoor subdued light and with good ventilation and observed in 2 out of 3 animals, then the dose administered
then crushed into a fine powder using a laboratory was concluded as toxic dose. No death was occurring up to
Homogenizer, which passed through 22 No sieve. the dose level of 500mg/kg.
The fruits of sativum were dried and then crushed into
fine powder by using laboratory homogenizer then stored Hepatoprotective Activity [14-21]:
for further use.
Group 1: Normal control, received 0.5% Carboxy methyl
Preparation of Plant Extracts cellulose (CMC) solution (1ml/kg) once daily for 7 Days.
Ethanol Extract of Cucumerina Linn. (EETC): Fine
powdered Leaves of cucumerina were extracted Group 2: Hepatotoxient, administered with paracetamol
successively with petroleum ether and Ethanol (60-80°C) (3gm/kg) a single dose on day 7.
using Soxhlet apparatus. The extract was filtered and
evaporated to separate solvent and residue. The semisolid
Group 3: Receives Trichosanthes cucumerina extract
residue thus obtained was stored in desiccator until further
(150mg/kg) once daily for 7 days. (EETC).
use.
Group 4: Receives combination of ethanol extract of
Ethanol Extract of Sativum: (EECS): Fine powdered
Trichosanthes cucumerina extract and coriandrum sativum
fruits of sativum were extracted successively with
extract in 1:1 ratio(150mg/kg) for 7 days.
petroleum ether and ethanol (60-80°C) using Soxhlet
MEETC&EECS.
apparatus in
Table 1: Effect of Extracts in Paracetamol induced hepatotoxicity rates
Parameters Control Paracetamol EETC EETC&EECS Standard
BILIRUBIN(d/l) 0.31±0.17 1.89±0.12*** 0.79±0.13*** 0.35±0.27*** 0.34±0.16***
SGOT (U/l) 135.03±12.21 419.65±25.93*** 218.83±14.12*** 195±14.75*** 171.06±17.75***
SGPT (U/l) 63.73±10.94 282.58±10.31*** 138.11±10.96*** 79.5±17.78*** 68.31±7.44***
ALP (U/l) 170.33±22.22 436.51±27.07*** 226.10±17.39*** 198.5±30.02*** 175.88±22.84***
*p < 0.05; ** p < 0.01; *** p < 0.001.
Values are expressed as mean (standard deviation; SD). Statistical significance was calculated with ANOVA followed by tukey compare all pair of coloum
treated group with paracetamol group


Group 5: Standard drug control, receives silymarin (100 mg/kg) once
daily for 7 days (Std ).
Group 3 to Group 4 receives paracetamol (3gm/kg ) as a single
dose on day 3, Thirty minutes after administration of drug extract and
silymarin respectively.

Assessment of Hepatoprotective Activity: All the extracts and the
paracetamol were administered orally by suspending in 0.5% CMC
solution. Animals were


1455

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