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. 2018 Sep;13(3):217–222. doi: 10.26574/maedica.2018.13.3.217

Hepatoprotective Role of Eclipta alba against High Fatty Diet Treated Experimental Models – A Histopathological Study

K SATHEESH NAIK 1, M GURUSHANTHAIAH 2, M KAVIMANI 3, K PRABHU 4, S LOKANADHAM 5
PMCID: PMC6290179  PMID: 31490461

Abstract

Background:

The hepatic injury due to oxidative stress was ameliorated through administration of an aqueous extract of Eclipta alba leaves and suggested that wedelolactone and demethylwedelolactone were the possible components of Eclipta alba behind the protective effect on liver as well as against liver disorders.

Objective:

To study the hepato-protective effects of Eclipta alba on high fatty diet treated experimental models.

Material and methods:

A total of 30 adult albino rats of Wistar strain weighing 165-215 grams, from the animal house of the Basaveshwara Medical College, Hospital and Research Centre, Chitradurga, were used for the present study: group 1 included animals fed with normal diet (control); group 2, animals treated with hyperlipidemic diet for eight weeks; group 3, animals treated with hyperlipidemic diet for eight weeks, followed by one week post treatment of Eclipta alba with normal diet; group 4, animals treated with hyperlipidemic diet for eight weeks, followed by two weeks post-treatment of Eclipta alba with normal diet; and group 5, animals treated with hyperlipidemic diet for eight weeks, followed by three weeks posttreatment of Eclipta alba with normal diet.

Results:

In animals with high fat diet (30%), we observed the deposition of fat in the form of fat lobules in and around the hepatocytes, mononuclear in filtration in the liver parenchyma, dilation of blood vessels, necrotic foci and damaged hepatocytes.

Conclusion:

The components of Eclipta alba like wedelolactone, demethylwedelolactone and saponins reduced fat deposition, mononuclear infiltration, and necrotic foci, and stimulated hepatocyte regeneration in the liver.

Keywords:high fat diet, mononuclear infiltration, necrotic foci, Eclipta alba, liver pathology.

INTRODUCTION

Non-alcoholic fatty liver disease (NAFLD) is one cause of a fatty liver, occurring when fat is deposited (steatosis) in the liver not due to excessive alcohol use. It is related to insulin resistance and the metabolic syndrome (obesity, combined hyperlipidemia, diabetes mellitus (type II) and high blood pressure) (1, 2). Non-alcoholic steatohepatitis (NASH) is the most extreme form of NAFLD and is regarded as a major cause of cirrhosis of the liver of unknown cause (3). Up to 80% of obese people have the disease (4). Non-alcoholic fatty liver disease is also more common among men than women in all age groups until the age of 60, where the prevalence between sexes becomes equal. This is due to the protective nature of estrogen (5). Eclipta alba L. (syn Eclipta erecta L. and Eclipta prostata L.) belongs to the Asteraceae family. This small annual herb with white/Yellow flowers spontaneously occurs in wet locations in South India. Wedelolactone (WL) and demethylwedelolactone (DWL) are the coumestans present in E. alba; they display a hepatocyto-protective action, enabling the regeneration of liver cells (6-15). The present study was undertaken to study the hepato-protective role of Eclipta alba against high fatty diet treated animals.

MATERIAL AND METHODS

A total of 30 adult albino rats of Wistar strain weighing 165-215 grams were used for the present study. All animals were procured from the animal house of the Basaveshwara Medical College, Hospital and Research Centre, Chitradurga. Animals were maintained as per the national guidelines and protocols approved by the institutional animal ethical committee dated 04/06/2015 and BMCH/IAEC/01 Anat/2015. The rats were maintained in laboratory under controlled environmental conditions (12 h light/dark cycle and room temperature (22-24° C), humidity (50+5%), and housed in polypropylene cages and given food and water ad libitum. The animals were fed with standard rat pellet diet commercially available and manufactured by the National Institute of Nutrition, Hyderabad and clean drinking water ad libitum. Rats were divided into five groups, each consisting of six animals.

Preparation of Extract

Dried roots of Eclipta alba were charged to a Soxhalate apparatus along with water. Extracts were obtained by heating the mass for 5-6 hours at a temperature of 60° C. This process was repeated. Extracts were then combined and filtered. The resulted product was charged to a drier unit to convert it into a powder form, which was further powdered in a multimill to a fine mesh size. Afterwards, it was sieved using a Sifter, to obtain uniform particle sizes, and then mixed in a blender to become uniform and homogenous. It was packed in food grade, virgin, double polythene bags, and the whole process was repeated.

Preparation of hyperlipidemic diet

For 1 kg of diet, carbohydrate 520 g, proteins 180 g, fats 300 g, 2% NaCl and 1% multivitamins were taken (15).

Animal grouping

Group 1: Animal fed with normal diet (Control). Group 2: Animals treated with hyperlipidemic diet for eight weeks. Group 3: Animals treated with hyperlipidemic diet for eight weeks, followed by one week post-treatment of Eclipta alba with normal diet. Group 4: Animals treated with hyperlipidemic diet for eight weeks, followed by two weeks post-treatment of Eclipta alba with normal diet. Group 5: Animals treated with hyperlipidemic diet for eight weeks, followed by three weeks post-treatment of Eclipta alba with normal diet

All animals were sacrificed after 12 hours of fasting on the last day of experimental study, and their liver was dissected and preserved in 10% formalin for 48 hours. Liver specimens were processed for routine histopathological examination to study the structural changes in liver of obese Albino rats fed on high fat diet.

RESULTS

On histopathological examination, animals in the control group showed hexagonal hepatic lobules with central vein, portal triad, Kupffer cells and radiating cords of polyhedral hepatocytes, sinusoids in liver (Fig. 1).

Histopathological changes in experimental group animals

Animals treated with hyperlipidemic diet for eight weeks (Group 2) showed dilated central vein, deposition of adipocytes more in the periportal area, scattered in between the hepatocytes, fat lobules and degenerated hepatocytes in the liver parenchyma (Fig. 2), deposition of macrovescicular fat, mononuclear infiltration around the portal triad, dilated sinusoids, numerous Kupffer cells and necrotic foci in the liver (Fig. 3).

Animals treated with hyperlipidemic diet for eight weeks, followed by one week post treatment of Eclipta alba with normal diet (Group 3) showed less dilation of the central vein, deposition microvescicular fat, some degenerated hepatocytes and few fatty lobules (Fig. 4), few Kupffer cells, less dilation of sinusoids and necrotic foci in the liver (Fig. 5) compared to animals fed with high fat diet.

Animals treated with hyperlipidemic diet for eight weeks, followed by two weeks post-treatment of Eclipta alba with normal diet (Group 4) showed few adipocytes around the central vein and in between the hepatocytes (Fig. 6), very few Kupffer cells in the liver (Fig. 7) and fewer degenerated hepatocytes, and necrotic foci, sinusoids and less mononuclear infiltration in the liver compared to animals in Groups 2 and 3.

In animals treated with hyperlipidemic diet for eight weeks, followed by three weeks posttreatment of Eclipta alba with normal diet (Group 5) we observed normal hepatocytes, portal triad, normal central vein, portal triad, few mononuclear infiltration, normal Kupffer cells and sinusoids in the liver (Fig. 8) as compared to groups 2, 3 and 4.

FIGURE 1.

FIGURE 1.

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FIGURE 1. Hexagonal hepatic lobules with central vein and portal triad in animals fed with normal diet (Group 1)

FIGURE 2.

FIGURE 2.

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FIGURE 2. Kupffer cells, central vein, radiating cords of polyhedral hepatocytes and sinusoids, under high magnification, in control group animals (40x) (Group 1)

FIGURE 3.

FIGURE 3.

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FIGURE 3. Deposition of fat in the periportal area and in between the hepatocytes, dilated central vein and fat lobules in the liver of animals treated with hyperlipidemic diet, under low magnification (10x) (Group 2)

FIGURE 4.

FIGURE 4.

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FIGURE 4. Deposition of macrovescicular fat in the periportal area, mononuclear infiltration around the portal triad, dilated sinusoids, numerous Kupffer cells and necrotic foci in the liver of animals treated with hyperlipidemic diet, under high magnification (40x) (Group 2)

FIGURE 5.

FIGURE 5.

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FIGURE 5. Deposition of adipocytes, degenerated hepatocytes, and necrotic foci in the liver of animals treated with hyperlipidemic diet, under high magnification (40x) (Group 2)

FIGURE 6.

FIGURE 6.

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FIGURE 6. Hyperlipidemic diet for eight weeks, followed by two weeks post-treatment of Eclipta alba animals, showed few adipocytes around the central vein and in between hepatocytes in the liver, under low magnification (10x) (Group 4)

FIGURE 7.

FIGURE 7.

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FIGURE 7. Hyperlipidemic diet for eight weeks, followed by two weeks post-treatment of Eclipta alba animals, showed very few Kupffer cell, fewer degenerated hepatocytes, necrotic foci, sinusoids and less mononuclear infiltration in the liver, under high magnification (40x) (Group 4).

FIGURE 8.

FIGURE 8.

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FIGURE 8. Hyperlipidemic diet for eight weeks, followed by two weeks post-treatment of Eclipta alba animals, showed very few Kupffer cell, fewer degenerated hepatocytes, necrotic foci, sinusoids and less mononuclear infiltration in the liver, under high magnification (40x) (Group 4).

DISCUSSION

Histological analysis revealed that high fat diet fed rats developed pathologic abnormalities in the liver and adipose tissue consistent with metabolic order. Chronic low-grade systemic inflammation is now considered to be a key component of metabolic order with white adipose known to secrete many relevant adipokines (16, 17). Visceral, not subcutaneous fat is associated with metabolic order in humans and rodents (18, 19). In our study, rats fed with normal diet had healthy livers, with no steatosis or inflammation and histologically normal hepatic cords and other architecture. Hepatosteatosis, formation of macrovescicular fat lobules, mononuclear infiltration, dilation of vessels, numerous Kupffer cells and necrosis occurred in the livers of animals fed with high fatty diet. This is in agreement with the previous literature; there was an increased adipose tissue, glucose intolerance, elevation of circulating serum lipids (20, 21). In this research, rats fed with high fat diet and treated with Eclipta alba for one week showed less dilation of the central vein, deposition of microvesicular fat, some degenerated hepatocytes and few fatty lobules, few Kupffer cells, less dilation of sinusoids and necrotic foci in the liver compared to animals with high fat diet. Rats that received a high fat diet and treatment with Eclipta alba for two weeks had few adipocytes around the central vein and in between the hepatocytes, Kupffer cells in the liver, few degenerated hepatocytes, and necrotic foci, sinusoids, less mononuclear infiltration in the liver compared to animals in Groups 2 and 3. Rats with high fat diet, treated with Eclipta alba for three weeks, had normal hepatocytes, portal triad, normal central vein, portal triad, few mononuclear infiltration, normal Kupffer cells and sinusoids in the liver when compared to Groups 2, 3 and 4. Eclipta alba root extract is considered a powerful liver tonic. It possesses a wide range of biological activities and is used for the treatment of hepatitis and cirrhosis (Wagner H, 1986) (22). A study conducted by Murugaian P et al in 2008, on the whole plant extract of Eclipta alba Hassk., exhibited a protective activity against CCl4 induced liver injury. The plant contains an alkaloid, ecliptine, which has a choleretic action and lipid peroxidation; the extract augmented bile flow in rats, suggesting a stimulation of the liver secretory capacity (23). Abraham P et al mentioned the same finding in their study on necrotic changes in hepatocytes (24). We detected mononuclear cell infiltrations between hepatocytes plates and around the portal triad. Regarding the underlying reason of this condition, some researchers claim that this inflammation area, either in foci or in a diffuse form, occurs due to cytokine release resulting from fatty feeding (25). E. alba was reported to be the best drug for treating liver cirrhosis and infective hepatitis; wedelolactone, demethylwedelolactone and saponins are believed to be the active principles responsible for the use of these drugs in liver disorders (26-36). These encouraging findings could portend extrapolation to human beings.

CONCLUSION

The present study reveals that Eclipta alba has a protective role against liver diseases such as liver cirrhosis and infective hepatitis. The components of Eclipta alba extract (wedelolactone, demethylwedelolactone and saponins) reduced fat deposition, mononuclear infiltration, necrotic foci and stimulated the regeneration of hepatocytes in the liver.

Conflicts of interest: none declared.

Financial support: none declared.

Acknowledgments: Authors are thankful to the Faculty and supporting staff of the Department of Anatomy, Basaveshwara Medical College, Chitradurga for their constant support during this work.

Contributor Information

K. SATHEESH NAIK, Department of Anatomy, Bharath University (BIHER), Chennai, Tamilnadu, India

M. GURUSHANTHAIAH, Department of Anatomy, Basaveshwara Medical College, Chitradurga, Karnataka, India

M. KAVIMANI, Department of Anatomy, Sree Balaji Medical College and Hospital, Chennai, Tamilnadu, India

K. PRABHU, Department of Anatomy, Sree Balaji Medical College and Hospital, Chennai, Tamilnadu, India

S. LOKANADHAM, Department of Anatomy, Santhiram Medical College and Hospital, Nandyal, Andhra Pradesh, India

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