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. 2014 Apr 23;21(5):409–416. doi: 10.1016/j.sjbs.2014.04.001

Hepatoprotective effect of Rhodiola imbricata rhizome against paracetamol-induced liver toxicity in rats

Ravichandran Senthilkumar 1, Rahul Chandran 1, Thangaraj Parimelazhagan 1,⁎
PMCID: PMC4191600  PMID: 25313275

Abstract

Rhodiola imbricata is a perennial herb of the family Crassulaceae, which has significant traditional usage as medicine and is also known to biosynthesize phytochemicals such as flavonoids, coumarins and phenyl glycosides. The present investigation was aimed to estimate the hepatoprotective activity of R. imbricata rhizome acetone extract against paracetamol (2 g/kg) induced liver toxicity. Paracetamol was administered to induce hepatic damage in Wistar rats. 200 and 400 mg/kg doses of rhizome acetone extract and silymarin (25 mg/kg) were used as treatment groups. The blood samples were analyzed for biochemical markers of hepatic injury and tissue samples were subjected for estimation of liver antioxidants and histopathological studies. Analysis of the extract treated rats (400 mg/kg) showed an elevation of superoxide dismutase (0.326 units/min/mg protein), catalase (185.03 μmole of H2O2 consumed/min/mg protein), glutothione peroxidase (19.26 mg GSH consumed/min/mg protein) and reduced glutathione (16.2 μmole of GSH/mg protein). Moreover, the biochemical parameters in serum like alkaline phosphatase, serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT) and lipid profiles were also improved in treated groups compared to the control. The oral administration of different doses of rhizome acetone extract significantly protected the hepatic cells from damage. The hematological and biochemical parameters were also normal in extract treated rats compared to the control and standard (silymarin) groups. The HPLC analysis revealed the presence of some important phenolic compounds which could be responsible for the hepatoprotective activity. This study proved that R. imbricata could be taken as a good natural source of the hepatoprotective agent.

Keywords: Hepatotoxicity, In vivo antioxidants, Histopathology, Hematology, Biochemical markers

1. Introduction

The liver as a vital organ in the body is primarily responsible for the metabolism of endogenous and exogenous agents. It plays an important role in drug elimination and detoxification and liver damage may be caused by xenobiotics, alcohol consumption, malnutrition, infection, anemia and medications (Mroueh et al., 2004). Paracetamol is widely used an analgesic and antipyretic, but produces acute liver damage at higher doses. The hepatotoxicity of paracetamol has been attributed to the formation of toxic highly reactive metabolite n-acetyl parabenzoquineimine (NAPQI). Despite the fact that hepatic problems are responsible for a significant number of liver transplantations and deaths recorded worldwide, available pharmacotherapeutic options for liver diseases are very limited and there is a great demand for the development of new effective drugs (Akindele et al., 2010). Conventional or synthetic drugs used in the treatment of liver diseases are inadequate and can have serious adverse effects. So there is a worldwide trend to go back to traditional medicinal plants. Many natural products of herbal origin are in use for the treatment of liver ailments (Mitra et al., 2000).

Rhodiola imbricata Edgew is a perennial herb of the family Crassulaceae, commonly known as golden or arctic root, grows on rocky slopes, common in drier areas of the western Himalaya at an altitude of 4000–5000 m. Rhodiola root has been used extensively since time immemorial for its medicinal properties in traditional folk medicine in China, Tibet, Mongolia and the former Soviet Republics to increase physical endurance, work productivity, longevity and to treat fatigue, asthma, hemorrhage, impotence and gastrointestinal ailments (Kelly, 2001). R. imbricata is the major constituent of herbal tea and is rich in antioxidant value. The aqueous extract of R. imbricata root was found to contain gallic acid, p-tyrosol, rosavin and rosin (Mishra et al., 2008). Recently, rhizome of R. imbricata, was found to possess radioprotective (Arora et al., 2005), cytoprotective and antioxidant (Kanupriya Prasad et al., 2005), wound healing (Gupta et al., 2007), immunomodulatory (Mishra et al., 2006), adaptogenic (Spasov et al., 2000), anti-fatigue (Darbinyan et al., 2000), neuroprotective (Mook-Jung et al., 2002) and antiproliferative activities in HT-29 human colon cancer cells (Senthilkumar et al., 2013). Increased concerns on the side effects of current therapeutic modalities, plants are being valued because of their efficient curative properties and least or no side effects. Hence an attempt has been made to assess the hepatoprotective role of R. imbricata.

2. Materials and methods

2.1. Collection and identification of plant material

The fresh rhizomes of R. imbricata were collected from the Western Himalayas, Leh-Ladakh, India during the month of August to September, 2011. The plant material was identified and authenticated by Dr. O.P. Chaurasia, an Ethnobotanist at the Defence Institute of High Altitude Research (DIHAR), Leh-Ladakh. A voucher specimen of the same is available at the Field Research Laboratory (FRL), Leh.

2.2. Preparation of plant extracts

Freshly collected plant materials were washed under running tap water and distilled water to remove adhering dust and then dried under shade. The dried samples were powdered in a mechanical grinder and used for solvent extraction.

2.3. Animals and management

Experiments were conducted using Wistar albino rats (male, 150–200 g) and Swiss albino mice (male, 25–30 g), procured from Small Animal Breeding Station (SABS), College of Veterinary and Animal Sciences, Mannuthy, Thrissur, Kerala. The animals were housed in groups for a minimum of 7 days prior to pharmacological experiments. Animal quarters were maintained at a temperature of 22 ± 2 °C and with a 12 h light/12 h dark cycle. The animals had free access to commercial food pellets and clean drinking water. The study received approval from the Institutes Animal Ethics Committee (IAEC) for the Committee (CPCSEA) for the Purpose of Control and Supervision of Experiments on Animals (Reg. No.KMCRET/Ph.D/04/2011).

2.4. Acute toxicity

An acute oral toxicity study was performed as per OECD guidelines for the testing of chemicals, Test No. 423 (OECD; acute oral toxicity-acute toxic class method). Swiss albino mice (n = 6) were used for the acute toxicity study. The animals were kept overnight with access to water but not food, after which the R. imbricata acetone extract was administered orally at a dose level of 500, 1000 and 2000 mg/kg body weight and the animals were observed for 24 h. Further, they were observed continuously for the first 2 h for morbidity and up to 24 h for mortality. If mortality was observed in 2 out of 3 animals, then the dose administered was identified as a toxic dose. If mortality was observed in one animal, then the same dose was repeated again to confirm the toxic dose. If mortality was observed again, the procedure was repeated for lower doses (300, 50 and 5 mg/kg body weight).

2.5. Hepatoprotective property of R. imbricata

Hepatotoxicity was induced by paracetamol induced liver damage model by Sreedevi et al. (2009). Paracetamol (Acetaminophen, Sigma Chemical Company, USA) was suspended in 0.5% Tween-80 and administered p.o., at a dose of 2 g/kg. Wistar albino rats (male) weighing between 150 and 200 g were divided into 5 groups of 6 animals each. The weight range of the animals was equally distributed throughout the groups. Group-I served as control and received water, Group-II served as negative control, administered with paracetamol (2 g/kg, p.o.), Group-III standard, silymarin received (Sigma Chemical Company, USA) at a dose of 25 mg/kg p.o., Group-IV and V received acetone extract (200 and 400 mg/kg, p.o., respectively), once daily for 14 days.

On the 14th day, blood samples were collected from all animals by puncturing retro-orbital plexus under mild ether anesthesia, later animals were sacrificed and liver tissues were collected. The blood samples were analyzed for biochemical markers of hepatic injury and tissue samples were subjected for estimation of liver antioxidants and histopathological studies.

2.5.1. Preparation of serum from blood

Blood was drawn by puncturing the retro-orbital plexus under diethyl ether anesthesia. Whole blood for hematogram was collected in bottles containing the anticoagulant, ethylene diamine tetra-acetic acid (EDTA) while samples for biochemical analysis were collected in plain sample bottles. Serum was separated by centrifugation at 600×g for 15 min. and analyzed for various biochemical parameters. Sera were stored in the −80 °C freezer before they were analyzed. The erythrocytes, leucocytes and platelets were determined with an Auto Hematology Analyzer (Sysmex F-800, Japan). Alkaline phosphatase (ALP), serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT) and lipid profiles like total cholesterol (TC), triglycerides (TG) and biochemical parameters such as creatinine and bilirubin were also determined using Span Diagnostics Limited kit, India.

2.5.2. Preparation of liver homogenate

Hepatic tissues were homogenized in 10% w/v 0.1 M phosphate buffer or 0.1 M tris buffer (pH 7.0) and centrifuged at 12,000×g for 10 min. The supernatant was used for the measurement of liver enzymatic and non enzymatic antioxidants.

2.5.3. Determination of antioxidant enzymes

Total Protein content of the tissues was estimated by the method of Lowry et al. (1951). Enzymatic antioxidants were determined by estimating superoxide dismutase (SOD) (Kakkar et al., 1984), catalase (CAT) (Sinha, 1972), glutothione peroxidase (GPx) activity (Rotruck et al., 1973) and non-enzymatic antioxidants by reduced glutathione (GSH) (Ellman, 1959) and lipid peroxidation (LPO) (Hogberg et al., 1974).

2.5.4. Histopathology

After the experimental period animals were sacrificed, liver removed immediately, sliced and washed in saline. Liver pieces were preserved in 10% formalin for histopathological studies. Pieces of the liver were processed and embedded in paraffin wax. Sections were taken and stained with hematoxylin and eosin and photographed.

2.6. High performance liquid chromatography (HPLC) analysis

HPLC analysis of acetone extract of R. imbricata was performed using the Shimadzu HPLC system. The HPLC system (Shimadzu, Japan) consisted of a LC-10AT VP pump, a SPD-10AVP, PDA detector, a Phenomenex Luna C18 (250 mm × 4.6 mm, 5 μm) column, a Phenomenex, HPLC guard cartridge system and a class cbm-20A/20 Alite software. Separation was performed in a reverse phase column by maintaining the isocratic flow rate (1 mL/min) of the mobile phase (methanol:water, 70:30) and peaks were detected at 226 nm. The standard substances gallic acid and rutin fractions were analyzed in the same conditions. Peaks were assigned by spiking the samples with standard substances and comparison of the retention time and UV spectrum.

2.7. Statistical analysis

The results were statistically analyzed and expressed as mean (n = 6) ± standard error. Values are analyzed by the Dunnet’s test (SPSS, ANNOVA statistical software 17.0, TULSA, USA).

3. Results

3.1. Acute toxicity

The acetone extract of R. imbricata was subjected to acute toxicity testing in Swiss albino mice and animals were monitored for 24 h. The acetone extract of R. imbricata rhizome did not cause any mortality up to 2000 mg/kg, and hence 1/10th and 1/5th of the maximum dose administered (i.e. 200 and 400 mg/kg, p.o.) were selected for the present study.

3.2. Effect of acetone extract of R. imbricata on hematological parameters in paracetamol intoxicated rats

The effect of acetone extract of R. imbricata at two dose levels (200 mg/kg and 400 mg/kg, p.o.) on hematological parameters in paracetamol induced hepatic damage is shown in Table 1. Hepatic injury induced by paracetamol caused a decrease in erythrocytes (RBC count, hemoglobin, hematocrit, MCV, MCH, MCHC, RDW), leucocytes (total WBC count, Polymorphs, Lymphocytes) and platelets. Silymarin (25 mg/kg) was used as reference standard. Administration of acetone extract (400 mg/kg) resulted in increased levels of hematological (RBC count 6.5 ± 0.5 mill/cmm, total WBC count 5800 ± 2.0 cells/cmm and platelets 396 ± 2.0 thou/cmm) parameters.

Table 1.

Effect of acetone extract of R. imbricata on hematological parameters in rats treated with paracetamol.

Group-I (control) Group-II (paracetamol) Group-III (silymarin 25 mg/kg) Group-IV (200 mg/kg) Group-V (400 mg/kg)
Erythrocytes
RBC count (mill/cmm) 7.1 ± 0.2 3.1 ± 0.1⁎ 6.9 ± 1.2 5.4 ± 0.2⁎ 6.5 ± 0.5
Hemoglobin (gm%) 13.5 ± 0.3 7.2 ± 1.1⁎ 12.3 ± 2.1 11.6 ± 2.1 11.8 ± 1.2
Hematocrit (%) 32.1 ± 1.3 17.1 ± 2.1⁎ 31.4 ± 1.4 25.1 ± 0.3⁎ 29.5 ± 1.1
MCV (fl) 47.6 ± 0.1 54.7 ± 0.3⁎ 46.3 ± 1.6 30.4 ± 0.4⁎ 35.2 ± 2.1⁎
MCH (pg) 19.1 ± 0.6 23.1 ± 0.3 17.1 ± 2.0 17.3 ± 0.2 16.2 ± 1.2
MCHC (%) 38.3 ± 1.1 42.1 ± 2.2⁎ 36.3 ± 2.3 30.1 ± 0.5⁎ 32.1 ± 2.1⁎
RDW (%) 11.3 ± 1.4 13.6 ± 3.0 10.1 ± 2.1 8.1 ± 1.0 9.2 ± 1.0



Leucocytes
Total WBC count (cells/cmm) 7300 ± 1.0 2600 ± 2.0⁎ 6000 ± 1.0⁎ 5000 ± 1.0⁎ 5800 ± 2.0⁎
Polymorphs (%) 8 ± 1.1 10 ± 0.4 7 ± 1.1 6 ± 2.1 6.2 ± 1.2
Lymphocytes (%) 88 ± 0.1 90 ± 0.3⁎ 78 ± 1.1⁎ 68 ± 0.4⁎ 70 ± 0.3⁎
Platelets (thou/cmm) 524 ± 1.5 84 ± 2.1⁎ 404 ± 0.2⁎ 377 ± 1.0⁎ 396 ± 2.0⁎
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Data represent mean ± S.E.M (n = 6).

⁎

p < 0.05 compared to corresponding control.

3.3. Effect of acetone extract of R. imbricata on serum biochemical markers in paracetamol intoxicated rats

Table 2 shows the effect of acetone extract (200 mg/kg and 400 mg/kg) on serum biochemical markers in paracetamol induced hepatic damage. Hepatic injury causes elevated level of liver enzymes such as SGOT (serum glutamic oxaloacetic transaminase), SGPT (serum glutamic pyruvic transaminase) and ALP (alkaline phosphatase). Treatment with R. imbricata at 400 mg/kg revealed comparable activity with reference standard silymarin (25 mg/kg). Acetone extract decreased the liver markers SGPT (88.43 ± 0.3 U/l), SGOT (79.56 ± 0.3 U/l) and ALP (193.53 ± 0.3 U/l).

Table 2.

Effect of acetone extract of R. imbricata on serum biochemical parameters in rats treated with paracetamol.

Biochemical markers Group-I (control) Group-II (only paracetamol) Group-III (silymarin, 25 mg/kg) Group-IV (200 mg/kg) Group-V (400 mg/kg)
Creatinine (mg/dl) 0.68 ± 0.04 0.73 ± 0.01 0.69 ± 0.02 0.71 ± 0.01 0.70 ± 0.03
Bilirubin (mg/dl) 0.16 ± 0.01 0.17 ± 0.08 0.16 ± 0.01 0.17 ± 0.01 0.17 ± 0.02
00SGOT (U/l) 34.74 ± 0.1 157.36 ± 0.3⁎ 53.63 ± 0.3⁎ 121.82 ± 0.6⁎ 88.43 ± 0.3⁎
SGPT (U/l) 32.71 ± 0.6 148.52 ± 0.4⁎ 47.35 ± 0.2⁎ 117.37 ± 0.5⁎ 79.56 ± 0.3⁎
ALP (U/l) 158.48 ± 0.4 261.56 ± 0.3⁎ 167.68 ± 0.4⁎ 224.53 ± 0.3⁎ 193.53 ± 0.3⁎
Cholesterol (mg/dl) 54.45 ± 0.1 53.59 ± 0.2⁎ 54.68 ± 0.2⁎ 53.49 ± 0.2⁎ 54.64 ± 0.5⁎
Triglycerides (mg/dl) 53.45 ± 0.1 51.39 ± 0.4⁎ 53.71 ± 0.3⁎ 53.42 ± 0.3 53.56 ± 0.3⁎
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Data represent mean ± S.E.M (n = 6).

SGOT – serum glutamic oxaloacetic transaminase, ALP – alkaline phosphatase, SGPT – serum glutamic pyruvic transaminase.

⁎

p < 0.05 compared to the corresponding control.

3.4. Effect of acetone extract of R. imbricata on in vivo antioxidant activity in paracetamol intoxicated rats

Administration of paracetamol at the concentration of 2 g/kg to the Wistar rats caused an elevation of lipid peroxidation and decreased enzymatic antioxidants. The acetone extract of R. imbricata (400 mg/kg, p.o.) rhizome increased the total protein (0.185 μg/10 mg of liver tissue), enzymatic antioxidants SOD (0.326 units/min/mg protein), CAT (185.03 μmole of H2O2 consumed/min/mg protein) and GPx (19.26 mg GSH consumed/min/mg protein). The results are shown in Table 3.

Table 3.

Effect of acetone extract of R. imbricata on total protein, enzymatic and non-enzymatic antioxidants in rats treated with paracetamol.

Parameters Group-I (control) Group-II (only paracetamol) Group-III (silymarin, 25 mg/kg) Group-IV (200 mg/kg) Group-V (400 mg/kg)
Groups
Total protein (μg/10 mg of tissue) 0.251 0.085 0.236 0.105 0.185
SOD (units/min/mg protein) 0.473 0.191 0.501 0.242 0.326
CAT (μmole of H2O2 consumed/min/mg protein) 234.27 98.4 229.38 145.71 185.03
GPx (mg GSH consumed/min/ mg protein) 21.64 12.73 23.53 14.76 19.26
GSH (μmole of GSH/mg protein) 18.84 11.52 17.98 14.26 16.2
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SOD – superoxide dismutase, CAT – catalase, GPx – glutothione peroxidase, GSH – reduced glutathion.

3.5. Histopathological examination

The hepatoprotective effect of the R. imbricata acetone extract against paracetamol induced injury was confirmed by histopathological examination. The hepatocyte damage caused by paracetamol is shown in Fig. 1. Paracetamol treated rat liver showed portal tract inflammation with lymphocysts and showed early fibrosis of the perivenular region but the R. imbricata acetone extract protects the liver hepatocytes by preventing oxidation in liver cells.

Figure 1.

Figure 1

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Histopathological examination for the hepatoprotective activity of R. imbricata in paracetamol treated Wistar rats. (A) paracetamol treated rat liver showing portal tract inflammation with lymphocyst (original magnification 10×); (B) paracetamol treated rat liver showing early fibrosis of the perivenular region; (C) protective effect of the R. imbricata (400 mg/kg) in the liver of the tested rats. Liver showing mild dilatation of sinusoids (original magnification 40×).

3.6. HPLC analysis of R. imbricata acetone extract

Qualitative analysis of data obtained using HPLC confirms the presence of gallic acid and rutin in the acetone extract of R. imbricata. The retention times of the compounds (Figs. 2 and 3) present in the acetone extract (4.881 and 2.545) were eluted very close to the standards in the chromatograms of rutin (5.299) and gallic acid (2.855).

Figure 2.

Figure 2

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HPLC chromatogram for rutin and gallic acid.

Figure 3.

Figure 3

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HPLC chromatogram for acetone extract of R. imbricata.

4. Discussion

The primary function of the liver is to maintain body homeostasis, besides it plays a key role in metabolism, detoxification, and inflammatory response (Tacke et al., 2009). The liver is a versatile organ in the body concerned with regulation of internal chemical environment. Therefore, damage to the liver inflicted by a hepatotoxic agent is of grave consequence. Acetaminophen, also called as paracetamol or APAP (N-acetyl-p-aminophenol), is often considered as a safe painkiller drug, eventhough, overdoses of this drug lead to acute liver failure and hepatic cytolysis (Michaut et al., 2014). Hepatotoxic drugs such as d-galactosamine and acetaminophen reduces liver functional capacity, which leads to an accumulation of waste products such as ammonia in the blood (Mao et al., 2014). Paracetamol is a common analgesic and antipyretic agent, known to possess hepatotoxic effects in humans at very high doses. It has been used as a successful experimental model to evaluate the efficacy of hepatoprotective agents (Sreedevi et al., 2009). The mode of action of paracetamol on the liver is by covalent binding of its toxic metabolite, n-acetyl-p-benzoquinone-amine to the sulfhydryl group of protein resulting in cell necrosis and lipid peroxidation (Kapur et al., 1994). Due to liver injury caused by paracetamol overdose, the transport function of the hepatocytes gets disturbed resulting in the leakage of the plasma membrane (Zimmerman and Seeff, 1970), thus causing an increase in serum enzyme levels.

Administration of R. imbricata acetone extract at concentrations of 200 and 400 mg/kg, p.o., for 14 days resulted in a significant reduction of paracetamol induced elevation of serum enzyme markers, comparable to the effect of silymarin, the positive control used. Silymarin is a known hepatoprotective compound obtained from Silybum marianum. It is reported to have a protective effect on plasma membrane of hepatocytes (Ramellini and Meldolesi, 1976). Hepatic injury induced by paracetamol caused a decrease in erythrocytes (RBC count, hemoglobin, hematocrit, MCV, MCH, MCHC, RDW), leucocytes (total WBC count, Polymorphs, Lymphocytes) and platelets.

It is known that many toxic compounds accumulate in the liver where they are detoxified (Clarke and Clarke, 1977). Liver transaminases such as AST (aspartate transaminase) or SGOT (serum glutamic oxaloacetic transaminase), and ALT (alanine transaminase) or SGPT (serum glutamic pyruvic transaminase) have still remained the gold standards for the assessment of liver injury, and have been used as biomarkers of choice for decades (Howell et al., 2014). A study of liver function tests may therefore prove useful in assessing especially the toxic effects of medicinal plants on the liver. These tests involve mainly the determination of AST and ALT (Tilkian, 1979) and any marked necrosis of the liver cells leads to a significant rise of these enzymes in the blood serum. The lack of this effect on these liver enzymes shows that the extract is non-toxic to the hepatocytes. The results of enzymatic antioxidants such as SOD (superoxide dismutase), CAT (catalase), GPx (glutathione peroxidase) and non-enzymatic antioxidants such as GSH (reduced glutathione) and LPO (lipid peroxidation) indicate that the extract is non-toxic and safe. SGOT, SGPT and ALP (alkaline phosphatase) are important serum enzymes in the human liver and usually help to detect chronic liver diseases by monitoring their concentrations. The values were well comparable to the control group. Hepatic injury causes elevated levels of liver enzymes such as SGOT, SGPT and ALP. Treatment with R. imbricata at the concentration of 400 mg/kg revealed a comparable activity with the reference standard silymarin, a potent hepatoprotective drug. Administration of paracetamol at the concentration of 2 g/kg to the Wistar rats caused an elevation of lipid peroxidation and decreased enzymatic antioxidants. The acetone extract of R. imbricata rhizome increased the total protein and enzymatic antioxidants.

In this experiment, the R. imbricata acetone extract appears to be effective in reducing the paracetamol caused injury as observed from a significant reduction of paracetamol induced elevated serum enzyme levels. It was also noted that the histopathological damage induced by paracetamol was improved in rat liver treated with plant extract. This implies that concomitant administration of R. imbricata prevented hepatonecrotic changes induced by the toxic dose of paracetamol.

Hepatoprotective effect of R. imbricata was further confirmed by histopathological studies of the liver, which basically supported the results from the serum assays. Histopathological studies of the liver showed fatty changes, swelling and necrosis with loss of hepatocytes in paracetamol treated rats. R. imbricata treated groups showed regeneration of hepatocytes, normalization of fatty changes and necrosis of the liver. The maximum protection against hepatic damage was achieved with the acetone extract at a dose of 400 mg/kg. The histopathological observations of the liver of rats treated with R. imbricata showed a more or less normal architecture of the liver having reversed to a large extent, the hepatic lesions produced by paracetamol, almost comparable to the normal control groups.

R. imbricata species is reported to be a potent adaptogen (Gupta et al., 2008; Perfumi and Mattioli, 2007). A high content of bioactive compounds such as rosavin, rosin, p-tyrosol and gallic acid was reported in aqueous extract of R. imbricata species (Mishra et al., 2008). Qualitative analysis of data obtained using HPLC confirms the presence of gallic acid and rutin or its analogs in the acetone extract of R. imbricata. GC/MS analysis revealed that the methanolic extract of R. imbricata contains phytosterols, alkyl halide, phenols, and fatty acid esters, and is responsible for its antiradical properties (Tayade et al., 2013). Salidroside [2-(4-hydroxyphenyl) ethyl β-d-glucopyranoside (1)], was treated under oxidative stress induced by hydrogen peroxide in human erythrocytes. Flow cytometric analysis revealed that Salidroside protects human erythrocytes by its antioxidative activity, and caspase-3 inhibition in a dose-dependent fashion (Qian et al., 2012). Rhodiola possesses Salidroside and its metabolite p-tyrosol as a major phenolic compound. These compounds were determined by LC–MS/MS based method in rat liver tissues using paracetamol as the internal standard (Guo et al., 2014). Poly (ADP-ribose) polymerase (PARP) is a DNA repair enzyme, and is actively involved in cell apoptosis. Salidroside protects hematopoietic stem cells from oxidative stress by activating PARP1 (Li et al., 2014). These studies reveal that the extract is rich in these antioxidant compounds, which clearly support the antioxidant and other related pharmacological properties including hepatoprotective activity of Rhodiola. Hence, the phenolic compounds present in R. imbricata rhizome might be responsible for its observed hepatoprotective activity. Further experiments are underway to discern the molecular mechanism of phytoconstituents.

5. Conclusion

From the present study it is evident that the acetone extract of R. imbricata rhizome has no toxicity even at a higher dose of 2000 mg/kg. Liver antioxidant markers were elevated significantly, while the serum and lipid parameters were maintained at normal levels compared to control groups. Histopathological examinations of the liver showed that extract and silymarin have a protective role over the toxicity of paracetamol. The HPLC profiles of rhizome showed the presence of some important phenolic compounds which could be responsible for imparting protection to the liver of rats. Hence R. imbricata could be one of the best sources of natural hepatoprotective agents.

Acknowledgement

The authors are thankful to the DRDO-DIHAR, Govt. of India for the financial support.

Footnotes

Peer review under responsibility of King Saud University.

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