Pharmacological Evaluation of Livodac Tablet on Ethanol Induced Hepatotoxicity Models in Wistar Rats

 

Pradnya. N. Jagtap, Prachali Ramdas Chavan, Dr. Suhas shitole

Pune District Education Association’s, Seth Govind Raghunath Sable College of Pharmacy, Saswad 412301, Tal. Purandar, Dist. Pune, Maharashtra, India.

*Corresponding Author E-mail: prachalichavan@gmail.com

 

ABSTRACT:

The various pharmaceutical dosages are affected on human health after long time or repeated use. Ethanol induce the toxicity to liver to use in high dose. These study was describe the significant effect of livodac formulation on Ethanol induced liver cirrhosis in rats. The thirty six animal were grouped in six set (1) Normal controle, it kept at normal condition (2) Disease controle group contains 50% Ethanol (2ml/kg) (3) Standard group Silyamarin gives with 50% Ethanol (2ml/kg) (4) Test drug (100mg/kg) with 50% Ethanol (2ml/kg) (5) Test drug (200mg/kg) with 50% Ethanol (2ml/kg) (6) Test drug (400mg/kg) with 50% Ethanol (2ml/kg). The present study show result of three dosage of livodac formulation should be significantly reduce the level of SGOT, SGPT, Total bilirubin, Total protein, Alkaline phosphate in the induce hepatotoxic model. The given livodac formulation effectively used as hepatoprotective agent in the management of liver cirrhosis caused by to various used drugs.

 

KEYWORDS: Pharmaceutical dosages, Ethanol, Hepatoprotective, Liver cirrhosis, livodac.

 

 


INTRODUCTION:

Liver is a vital organ that plays a role in controlling critical biochemical and physiological activities including homeostasis, growth, energy and nutrient supply, detoxification of drugs and other xenobiotics, and also combating infections[1,2]. Therefore, it is very susceptible to being damaged by hepatotoxic agents[3]. The Liver injuries induced by various hepatotoxins have been recognized as a major toxicological problem for years. Liver Cirrhosis is one of the liver injury occurs due to the necrosis of liver cells by fibrosis and nodule formation. Impairment in function and structure of liver leads to impaired liver blood flow and function.[4]

 

Cirrhosis is defined as a diffuse hepatic process characterized by fibrosis and the conversion of normal liver architecture into structurally abnormal nodules. Liver cirrhosis has become one of the major causes of morbidity and mortality. The number of deaths caused by liver cirrhosis in the Caribbean, Latin America, Asia, Oceania, Africa and Europe had increased significantly from 1980 to 2010.[5] Plants are a rich source used for centuries to cure various diseasesand disorders from natural products. Hence, it is necessary to find out new formulation from natural sources that have less expensive, least side effects, on a long term therapy which can provide better safety and efficacy.

 

However, there are a number of herbal formulations available on liver disorders in ayurvedic medicine.[6] Various formulations like Liv-52,[7]  Kamilari,[8] APCL-A AmalkadiGhrita,[9] and PanchagvyaGhrita, Himoliv,[10] are well known for their hepatoprotective effects. Polyherbal combination contain different herbal plant, they have as different pharmacological activities, the plant like terminalia chebula, terminalia belerica, emblica officinalis, azadiractaindica, swertiachirata, picrorhizakurroo, adhatodavasica, tinosporacardifolia.

 

MATERIAL AND METHODS:

Material:

The various crude drugs Terminalia chebula, Terminalia bellerica, Emblica officinalis, Azadiracta indica, Swertia chirata, Picrorhiza kurroo, Adhatoda vasica, Tinospora cardifolia are used in livodac formulation.

 

Methods:

1.     Preparation of polyherbal formulation:

All herbal drugs are give mix well by using wet granulation method and prepare the tablet by using direct compression method.

 

2.     In vivo model:

a.     Acute oral toxicity study:

The acute toxicity study of livodac formulation was performed by using mice either sex. It carryout according to OECD guideline 425.In that the animal should be fasted at over night and the dosing should be start in two group in first group give 2000mg/kg dose, and second group 5000mg/kg.

 

In 2000mg/kg give one animal dosing it is survive, so the addition of four animal in group the total five animal in that group are studded in that no death observed. These group are observed upto 14 days. No any death should beobserved the 2000mg/kg dose selected its 1/10th part should be used for test drug.

 

In 5000mg/kg Dose one animal at the test dose the animal dies.

 

b.    Procedure:

Select the animal 200-250gm Wister rats of either sex divided animal in six group (normal, disease control, standard, sub therapeutic, super therapeutic, therapeutic) weight of all animal before (day 1), during (day 7) and after (day 14) of the drug treatment.

 

After completion of the treatment, animals were anaesthetized under by chloroform anesthesia. Blood was collected by retro-orbital plexus puncture. Biochemical estimations of various parameters such as total blood count, total proteins, total Bilirubin, serum glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), and alkaline phosphatase (ALP) using serum were assessed. Animals were sacrificed, liver and kidney was removed, weighed and perfused in 10% formalin solution for histopathology studies.

 

Evaluation Parameter:

The rats were sacrificed on the eleventh day by using chloroform and blood was collected in plain tubes. The various blood tests were done SGOT, SGPT, Total Bilirubin, Total Alkaline Phosphate, Total protein.

 

Statistical Analysis:

The data were statistically analyzed and all values were expressed as Mean ±SEM. The data were also analyzed by One Way ANOVA using Graph pad prism 8 version Software.

 


Group No.

Treatment

No. of animal

I

Normal control: regular diet and drinking water

6

II

Positive control - 50% Ethanol (2ml/kg)

6

III

Standard control: Silyamarin (54mg/kg body weight) +50% Ethanol (2ml/kg)

6

IV

Sub therapeutic -test drug (100mg/kg body weight) + 50% Ethanol (2ml/kg)

6

V

Therapeutic - test drug (200mg/kg body weight) +50% Ethanol (2ml/kg)

6

VI

Super therapeutic - test drug (400mg/kgbody weight) +50% Ethanol (2ml/kg)

6

 


Table 1. Effect of livodac tablet on total Bilirubin, alkaline phosphate, SGOT, SGPT and Total protein on day 15th in Ethanol induced model.

Groups

Bilirubin

ALP

SGOT

Total Protein

SGPT

NC

112.31±18.27

0.54±0.16

33.01±2.23

7.16±0.19

32.76±1.16

DC

292.53±16.97###

0.96±0.07###

39.98±2.39#

8.49±0.46

44.35±0.98###

STD

122.84±8.36***

0.43±0.06***

29.98±1.27***

7.04±0.24*

31.33±0.67***

T100

222.84±10.46*

0.35±0.03***

31.03±0.92**

5.32±0.25***

25.68±1.21***

T200

239.63±22.30

0.46±0.04***

27.71±1.29***

5.26±0.54***

28.08±1.12***

T400

193.59±8.14***

0.38±0.02***

27.63±0.87***

6.19±0.41***

29.03±1.71***

 

 

Fig.1. Effect of livodac tablet on total Bilirubin

Fig.2. Effect of livodac tablet on Alkaline phosphate

Fig.3. Effect of livodac tablet on protein

Fig.4. Effect of livodac tablet on SGOT total

Fig.5. Effect of livodac tablet on SGPT

 


Table 2. Effect of livodac tablet on body weight in ethanol induced model.

Groups

NC

DC

STD

T100

T200

T400

Day1

245.5±2.21

246.66±1.66

243.33±2.10

242.5±3.09

245±1.82

246.66±1.58

Day7

233.66±6.83

278.5±6.23###

256.66±7.26

263.5±5.53

254.33±3.37*

258.83±5.31

Day15

236.16±5.02

288.5±5.00###

269.16±8.30

261.83±6.35*

258.83±6.87**

259.16±4.19**

 

Fig. 7.5.12 Data of body weight on day 1st

fig7.5.13 data of body weight on day 7th

Fig.7.5.14 data of body weight on day 15th

Fig 7.10.1Group 1: NC

Pathological change: NAD

Fig 7.10.2Group 2: DC

Pathological change: Minimal (+)

Fig. 7.10.3 Group 3: STD

Pathological change: Minimal (++)

Fig 7.10.2Group 2: DC

Pathological change: Minimal (+)

Fig. 7.10.3 Group 3: STD

Pathological change: Mild (++)

Fig. 7.10.4Group 4: Test (subtherapeutic)

Pathological Change: Moderate (+++)

Fig. 7.10.5Group5:Test(Therapeutic)

Pathological change: Minimal (+)

Fig. 7.10.6 Group6:Test (Supertherapeutic)

Pathological change: Mild (++)

Fig. 7.10.7 Group 1: NC

Pathological change: NAD

Fig.7.10.8 Group2: DC

Pathological change: Minimal (+)

Fig. 7.10.9Group 3: STD Pathological

change: Minimal (+)

Fig. 7.10.10Group 4: Test (sub therapeutic)

Pathological Change: Mild (++)

Fig. 7.10.11 Group 5: Test (Therapeutic)

Pathological change: Minimal (+)

Fig. 7.10.11Group 6: Test (Supertherapeutic)

Pathological change: Mild (++)

Fig. 3 Histopathology of liver

 


DISCUSSION:

The objective of these study was to ethanol induce the liver damage by their oxidation property and the livodac gives preventive effect by their antioxidant property.

 

The oral administration of ethanol at dose50% Ethanol (2ml/kg) was found to significantly alter the blood parameters like total bilirubin, SGOT, SGPT, total protein and alkaline phosphate.[11]

 

Ethanol was absorbed from the jejunum (the major site), and small amounts of fat are also absorbed from the mouth, esophageal, gastric, and large intestine mucosal membranes. Approximately 90% of ingested alcohol was metabolized in the liver. The Alcohol was primarilyoxidized to acetaldehyde by ADH in the cytosol of hepatocytes, and partly metabolized by cytochrome P-450 and catalase in the hepatocyte microsomes and hepatocyte peroxisomes, respectively. Acetaldehyde dehydrogenase converts acetaldehyde to acetate primarily in the hepatocyte mitochondria.[12]

 

Acetaldehyde was the key toxin in alcohol-induced liver injury, causing cellular damage, inflammation, extracellular matrix remodeling, and fibro genesis to cirrhosis.[13,14]

 

The varius parameters changed in various group are compaired with the disease controle group. In that study the six groups are given first normal control, diease control, standard, test drug with subtherapeutic dose, therapeutic dose and supertherapeutic dose (find by toxicity study). The all groups are compairedwith the disease controle group.

 

The total bilirubin level is significantly decreased (P<0.001) in comparison with the disease controle animals. The alkaline phosphate level in all groups are the significantly decreased (P<0.001) without the sub therapeutic group it gives compare with the disease controle group. Although in the SGOT the normal controle group (P<0.002). Total protein content the subtherapeutic and therapeutic gives the more significant effect (P<0.001).

 

The body weight on the 7th day gives change but the livodac gives to prevent the liver damage as significant (P<0.001), In the standard and the subtherapeutic they gives to less significant.

 

The histopathology of liver and kidney was lastly done it gives the mild (++) to moderate (++) change in liver and also in kidney observe. Mainly in the disease controle and the subtherapeutic group the moderate changes are observed.

 

 

CONCLUSION:

In this study the livodac contains various hepatoprotective plants having vast therapeutic potential as per reports. The present study showed that treatment of the livodac formulation at 200mg/kg dose in Ethanol induced rats showed hepatoprotective effect. Based on the above results it can be concluded that the livodac may act as hepatoprotective agent in experimental rats at the given dose.

 

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11.   F. yahya. et al. hepatoprotective activity of methanolic extract of bauhinia purpurea leaves against paracetamol induced hepatic damage in rats. Evidence Based and Complimentary Alternative Medicine. 2013,1-10

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Received on 07.02.2020         Modified on 10.03.2020

Accepted on 09.04.2020       ©A&V Publications All right reserved

Res.  J. Pharmacology and Pharmacodynamics.2020; 12(2): 91-96.

DOI: 10.5958/2321-5836.2020.00018.X