Hypolipidemic Activity of <em>Cassia tora</em> Seeds in Hyperlipidemic Rats
Abstract
The hypolipidemic activity of Cassia tora (Chakvat, Chakunda) (Family: Caesalpiniaceae) seeds extract have been studied in two models of hyperlipidemia in rats. In an acute model, hyperlipidemia was induced by injecting a single dose of Triton WR-1339 (400 mg/kg, b.w.) intraperitonially in rats. Feeding with C. tora seed extract at the dose of 500 mg/kg, b.w. exerted significant lipid lowering effect as assessed by the reversal of plasma levels of total cholesterol, phospholipids, triglyceride and reactivation of post heparin lipolytic activity. In the chronic model, hyperlipidemia was induced by feeding with cholesterol rich-HFD in rats. The treatment with seeds extract of C. tora (500 mg/kg, b.w.) simultaneously for 15 days also caused lowering of lipid levels in plasma and liver following reactivation of plasma post heparin lipolytic activity and hepatic lipoprotein lipase activity in animals. The hypolipidemic activity of C. tora seeds was compared with a standard drug guggulipid (200 mg/kg, b.w.) in both models.
Introduction
Cassia tora Linn (Family: Caesalpiniaceae) commonly known Chakvat, Chakunda and Charota in Hindi, Foetid Cassia in English is an herbaceous foetid annual weed, almost on under shrub, up to 90 cm in height. It grows in tropical and Asian countries especially on way sides and waste places and on hills of low elevations up to 1,800 m as well as in plains. Different parts of the plant (Leaves, seed, and root) are claimed to be effective against a variety of ailments in indigenous medicine [1]. The leaves and seeds are acrid, thermogenic, laxative depurative, antiperiodic, liver tonic, antihelmintic, cardio tonic and are useful in ringworm, pruritis, leprosy, skin disease, jaundice, helminthiasis, flatulence, dyspepsia, intermittent fevers, constipation, ophthalmopathy, cough, bronchitis, cardiac disorders and haemorrhoids [2, 3]. The leaves of C. tora are reported to have antirheumatic activity in folklore practice. Decoction of the leaves is used as laxative. The seeds of C. tora have been used in Chinese medicine as vision-improving, cardiotonic, hypolipidemic, aperients, antiasthnic and diuretic agent. Several polyharbal formulations are available in Chinese market for preventing the formation of atherosclerosis plaque [4].
Cardiovascular diseases are leading cause of death in both industrialized and developing nations [5]. Disorders of lipid metabolism following oxidative stress are the prime risk factors for initiation and progression of heart diseases [6]. The current therapies used for controlling hyperlipidemia; fibrates, stains and bile acid sequestraints are almost inefficient to regulate lipid metabolism. Furthermore, these drugs also cause a number of serious adverse effects in patients. Currently available treatment for hyperlipidemia in modern medicine, fibrats, statins or bile acids sequestraints and their combinations do not regulate lipid metabolism up to a appreciable mark, also have several adverse effects in patients [7]. Therefore, there is a need to develop safe and effective treatment modalities for hyperlipidemia. Further more medicinal plants play an important role in the treatment of lipid disorders, especially due to their lesser toxicity, side effects and cost effectiveness. Therefore, the research and development of lipid lowering drugs from natural products are the best option and also are in great demand. In view of the above considerations, the present study was designed to investigate lipid lowering activity of C. tora seeds in hyperlipidemic rats.
Materials and Methods
Preparation of Seed Extract
Cassia tora seeds were collected from local area of Lucknow and identified taxonomically by Department of Pharmacology, Era’s Lucknow Medical College Lucknow. A voucher specimen (CT-005/10) was also submitted. Seeds were crushed and dried under shade. The powder (500 g) was extracted with 95 % ethanol in a soxhlet extractor for 72 h, the extract was concentrated to dryness under reduced pressure and controlled temperature (50–60 °C), yielding 23 g of reddish brown solid (crude extract). This was stored in refrigerator and used to investigate hypolipidemic activity in rats. Triton WR-1339, deoxycholic acid, cholesterol and heparin were procured from Sigma Chemical Company, St. Louis, MO, USA. Guggulipid, a potent lipid lowering agent from gum resin of Commephora mukul (guggul) developed in Central Drug Research Institute, Lucknow, was used as a standard drug [8].
Preparation of Cholesterol Rich-High Fat Diet
Deoxycholic acid (5 g) was mixed thoroughly with 700 g of powdered rat chow diet supplied by Ashirvad Industries, Chandigarh, India. Simultaneously cholesterol (5 g) was dissolved in 300 g warm coconut oil. This oil solution of cholesterol was added slowly into powdered mixture to obtain homogeneous soft cake. This cholesterol rich-high fat diet (HFD) was molded in shape of pellet of about 3 g each [9].
Animals
In vivo experiments were conducted as per guidelines provided by Animal Ethics Committee of Central Drug Research Institute, Lucknow, India. Male adult rats of Charles Foster strain (200–225 g) bred in animal house of the Institute were used. The animals were housed in polypropylene cages and kept in uniform hygienic conditions, temperature 25–26 °C, relative humidity 50–60 % and 12/12 h light/dark cycle (light from 8:00 a m to 8:00 p m) and provided with standard rat pellet diet and water ad libitum.
Triton and Cholesterol Rich-HFD Induced Hyperlipidemia
The rats were divided into four groups having six animals in each as follows: control, hyperlipidemic, hyperlipidemic treated with C. tora seeds or guggulipid. In the acute experiment to induce hyperlipidemia, Triton WR-1339 was administered (400 mg/kg, b.w., p.o.) by intraperitonial injection. C. tora seeds extract and guggulipid were macerated with aqueous gum acacia (1 % w/v) suspension and fed orally at the doses of 500 and 200 mg/kg, b.w., respectively, simultaneously with triton. Control animals received same amount of vehicle. The diet was withdrawn and blood from fasted rats was collected after 18 h. Animals were anaesthetized with thiopentone solution (50 mg/kg, b.w.,i.p.), prepared in normal saline. Heparin (10 mg/ml) in normal saline was prepared and injected to each rat (1 mg/kg, b.w.) through tail vein. After 15 min, blood was withdrawn from retro-orbital plexus using glass capillary in EDTA coated tubes. In the chronic experiment, hyperlipidemia was produced by feeding with cholesterol rich-HFD for 15 days. Drugs were administered orally once daily at the same doses as above, simultaneously, with cholesterol rich-HFD in the drug treated groups. Control animals, kept over normal rat pellet diet, received the same amount of vehicle. At the end of experiment, rats were fasted overnight and anaesthetized. Blood was withdrawn just after 15 min of heparin treatment. Thereafter animals were sacrificed, liver was excised promptly, washed with cold 0.15 M KCl and kept it −40 °C till analyses. Blood was centrifuged and plasma was taken.
Biochemical Analysis of Plasma and Liver
Plasma Post heparin lipolytic activity (PHLA) was assayed in plasma spectrophotometrically using intralipid as artificial substrate [10] plasma was diluted with normal saline in a ratio of 1:3 and used for the analysis of total cholesterol (TC), phospholipids (PL) and triglyceride (TG) using standard enzymatic kits supplied by Merck India Ltd. Mumbai India [11–13]. Liver was homogenized (10 %w/v) in cold 100 mM phosphate buffer pH 7.2 and used for the assay of lipoprotein lipase (LPL) activity [10]. The lipid extract of each homogenate prepared in a mixture of CHCl3:CH3OH (2:1, v/v) was used for estimation of TC, PL and TG. Plasma and tissue were also estimated for protein content [14].
Statistical Analysis
One way analysis of variance (New man’s Student t test) was performed by comparison of values for hyperlipidemic groups with control, hyperlipidemic and drug treated groups with hyperlipidemic All hypothesis testing were two-tailed. P < 0.05 was considered statistically significant and results were expressed as mean ± SD of six rats. The graph pad INSTAT 3.0 software carried out the statistically analysis [15].
Preparation of Seed Extract
Cassia tora seeds were collected from local area of Lucknow and identified taxonomically by Department of Pharmacology, Era’s Lucknow Medical College Lucknow. A voucher specimen (CT-005/10) was also submitted. Seeds were crushed and dried under shade. The powder (500 g) was extracted with 95 % ethanol in a soxhlet extractor for 72 h, the extract was concentrated to dryness under reduced pressure and controlled temperature (50–60 °C), yielding 23 g of reddish brown solid (crude extract). This was stored in refrigerator and used to investigate hypolipidemic activity in rats. Triton WR-1339, deoxycholic acid, cholesterol and heparin were procured from Sigma Chemical Company, St. Louis, MO, USA. Guggulipid, a potent lipid lowering agent from gum resin of Commephora mukul (guggul) developed in Central Drug Research Institute, Lucknow, was used as a standard drug [8].
Preparation of Cholesterol Rich-High Fat Diet
Deoxycholic acid (5 g) was mixed thoroughly with 700 g of powdered rat chow diet supplied by Ashirvad Industries, Chandigarh, India. Simultaneously cholesterol (5 g) was dissolved in 300 g warm coconut oil. This oil solution of cholesterol was added slowly into powdered mixture to obtain homogeneous soft cake. This cholesterol rich-high fat diet (HFD) was molded in shape of pellet of about 3 g each [9].
Animals
In vivo experiments were conducted as per guidelines provided by Animal Ethics Committee of Central Drug Research Institute, Lucknow, India. Male adult rats of Charles Foster strain (200–225 g) bred in animal house of the Institute were used. The animals were housed in polypropylene cages and kept in uniform hygienic conditions, temperature 25–26 °C, relative humidity 50–60 % and 12/12 h light/dark cycle (light from 8:00 a m to 8:00 p m) and provided with standard rat pellet diet and water ad libitum.
Triton and Cholesterol Rich-HFD Induced Hyperlipidemia
The rats were divided into four groups having six animals in each as follows: control, hyperlipidemic, hyperlipidemic treated with C. tora seeds or guggulipid. In the acute experiment to induce hyperlipidemia, Triton WR-1339 was administered (400 mg/kg, b.w., p.o.) by intraperitonial injection. C. tora seeds extract and guggulipid were macerated with aqueous gum acacia (1 % w/v) suspension and fed orally at the doses of 500 and 200 mg/kg, b.w., respectively, simultaneously with triton. Control animals received same amount of vehicle. The diet was withdrawn and blood from fasted rats was collected after 18 h. Animals were anaesthetized with thiopentone solution (50 mg/kg, b.w.,i.p.), prepared in normal saline. Heparin (10 mg/ml) in normal saline was prepared and injected to each rat (1 mg/kg, b.w.) through tail vein. After 15 min, blood was withdrawn from retro-orbital plexus using glass capillary in EDTA coated tubes. In the chronic experiment, hyperlipidemia was produced by feeding with cholesterol rich-HFD for 15 days. Drugs were administered orally once daily at the same doses as above, simultaneously, with cholesterol rich-HFD in the drug treated groups. Control animals, kept over normal rat pellet diet, received the same amount of vehicle. At the end of experiment, rats were fasted overnight and anaesthetized. Blood was withdrawn just after 15 min of heparin treatment. Thereafter animals were sacrificed, liver was excised promptly, washed with cold 0.15 M KCl and kept it −40 °C till analyses. Blood was centrifuged and plasma was taken.
Biochemical Analysis of Plasma and Liver
Plasma Post heparin lipolytic activity (PHLA) was assayed in plasma spectrophotometrically using intralipid as artificial substrate [10] plasma was diluted with normal saline in a ratio of 1:3 and used for the analysis of total cholesterol (TC), phospholipids (PL) and triglyceride (TG) using standard enzymatic kits supplied by Merck India Ltd. Mumbai India [11–13]. Liver was homogenized (10 %w/v) in cold 100 mM phosphate buffer pH 7.2 and used for the assay of lipoprotein lipase (LPL) activity [10]. The lipid extract of each homogenate prepared in a mixture of CHCl3:CH3OH (2:1, v/v) was used for estimation of TC, PL and TG. Plasma and tissue were also estimated for protein content [14].
Statistical Analysis
One way analysis of variance (New man’s Student t test) was performed by comparison of values for hyperlipidemic groups with control, hyperlipidemic and drug treated groups with hyperlipidemic All hypothesis testing were two-tailed. P < 0.05 was considered statistically significant and results were expressed as mean ± SD of six rats. The graph pad INSTAT 3.0 software carried out the statistically analysis [15].
Results
Effect of C. tora Seeds Extract in Triton Induced Hyperlipidemia
The data in Table 1 shows that acute administration of Triton WR-1339 in rats caused marked increase in their plasma levels of TC (168 %), PL (187 %) and TG (176 %) following inhibition of PHLA by 31 %. Treatment with C. tora seeds extract at the dose of 500 mg/kg, b.w., caused decrease in these levels of TC, PL and TG by 28, 25 and 26 %, respectively, simultaneously with reactivation of PHLA by 22 %. However lipid lowering action of guggulipid even at a lower dose of 200 mg/kg, b.w., was comparatively higher (33–40 %) to that of C. tora seeds extract.
Table 1
Effect of Cassia tora seeds extract and guggulipid on plasma lipids in triton induced hyperlipidemia in rats
| Groups | Total cholesterol (mg/dl) | Phospholipid (mg/dl) | Triglyceride (mg/dl) | Post heparin lipolytic activity (n mole FFA released/h/L) |
|---|---|---|---|---|
| Control | 87.52 ± 6.11 | 82.46 ± 3.33 | 90.66 ± 8.69 | 16.86 ± 0.95 |
| Triton treated | 234.60*** ± 12.09 (+168) | 235.89*** ± 12.09 (+186) | 250.57*** ± 24.86 (+176) | 11.56** ± 1.34 (−31) |
| Triton + Cassia tora (500 mg/kg, b.w.) | 169.87** ± 6.14 (−28) | 177.89** ± 8.69 (−25) | 185.88** ± 12.80 (−26) | 14.11* ± 0.70 (+22) |
| Triton + Guggulipid (200 mg/kg, b.w.) | 153.13*** ± 8.65 (−35) | 159.14*** ± 7.85 (−33) | 149.14*** ± 8.23 (−40) | 14.36* ± 0.68 (+24) |
Values are mean ± SD of 6 animals. Values in parenthesis indicate percent change. Triton treated group is compared with control, triton and drug treated with triton
* P < 0.05; ** P < 0.01; *** P < 0.001
Effect of C. tora Seeds Extract in Cholesterol Rich-HFD Induced Hyperlipidemia
In this model of hyperlipidemia (Table (Table2),2), feeding with cholesterol rich-HFD in rats caused significant increase in their plasma levels of TC (135 %), PL (72 %) and TG (119 %) following inhibition of PHLA by 32 %. Treatment with C. tora seeds extract for 15 days, reversed these plasma levels of TC, PL and TG by 25, 21 and 31 %, respectively, simultaneously with reactivation of PHLA by 21. %. The hypolipedemic action of C. tora at the dose of 500 mg/kg, b.w., was comparable to that of guggulipid at the dose of 200 mg/kg, b.w. in above model.
Table 2
Effect of Cassia tora seeds extract and guggulipid on plasma lipids in cholesterol rich-HFD induced hyperlipidemia in rats
| Groups | Total cholesterol (mg/dl) | Phospholipid (mg/dl) | Triglyceride (mg/dl) | Post heparin lipolytic activity (n mole FFA released/h/L) |
|---|---|---|---|---|
| Control | 88.59 ± 6.98 | 80.62 ± 4.29 | 114.80 ± 11.30 | 19.31 ± 1.30 |
| Cholesterol rich-HFD treated | 208.18*** ± 22.10 (+135) | 138.27*** ± 24.86 (+72) | 251.35*** ± 24.59 (+119) | 13.11** ± 1.18 (−32) |
| Cholesterol rich-HFD +Cassia tora (500 mg/kg, b.w.) | 155.36** ± 11.13 (−25) | 108.86* ± 6.78 (−21) | 172.88** ± 6.49 (−31) | 15.88* ± 1.78 (+21) |
| Cholesterol rich-HFD + Guggulipid (200 mg/kg, b.w.) | 153.39** ± 8.83 (−26) | 100.30** ± 8.77 (−28) | 169.92** ± 6.09 (−32) | 16.33 * ± 1.64 (−25) |
Values are mean ± SD of 6 animals. Values in the parenthesis indicate percent change. Cholesterol rich-HFD treated groups is compared with control, cholesterol rich-HFD and drug treated groups with cholesterol rich-HFD
* P < 0.05; ** P < 0.01; *** P < 0.001
Effect of C. tora Seeds Extract in Cholesterol Rich-HFD Induced Steosis in Liver
Feeding with cholesterol rich-HFD in rats also caused accumulation of TC (53 %), PL (72 %) and TG (57 %) following diminution of LPL activity by 37 % in their liver (Table 3). However, treatment with C. tora seeds extract exerted a decrease in these levels of TC, PL and TG by 26, 22 and 29 %, respectively, following reactivation of LPL activity (26 %) in hyperlipidemic animals. Guggulipid was more effective hypolipedemic than C. tora seeds, as it could decrease the level of lipids by 26–40 %, following reactivation of LPL activity (29 %) in the liver of hyperlipidemic rats.
Table 3
Effect of Cassia tora seeds extract and guggulipid on Liver lipid in cholesterol rich-HFD induced hyperlipidemia in rats
| Groups | Total cholesterol (mg/dl) | Phospholipid (mg/dl) | Triglyceride (mg/dl) | Lipoprotein lipase activity (n mole FFA released/h/mg protein) |
|---|---|---|---|---|
| Control | 6.75 ± 0.30 | 22.43 ± 2.71 | 4.54 ± 0.38 | 79.85 ± 4.82 |
| Cholesterol rich-HFD treated | 10.33*** ± 0.62 (+55) | 38.58*** ± 5.33 (+72) | 8.56*** ± 1.02 (+87) | 50.26*** ± 2.72 (−37) |
| Cholesterol rich-HFD + Cassia tora (500 mg/kg, b.w.) | 7.67** ± 0.62 (−26) | 29.40** ± 3.60 (−24) | 5.22*** ± 0.74 (−39) | 63.09* ± 3.75 (+25) |
| Cholesterol rich-HFD +Guggulipid (200 mg/kg, b.w.) | 7.36** ± 0.90 (−29) | 28.47** ± 5.04 (−26) | 5.11*** ± 0.62 (−40) | 64.99** ± 4.66 (−29) |
Values are mean ± SD of 6 animals. Values in the parenthesis indicate percent change. Cholesterol rich-HFD treated groups is compared with control, cholesterol rich-HFD and drug treated groups with cholesterol rich-HFD
* P < 0.05; ** P < 0.01; *** P < 0.001
Effect of C. tora Seeds Extract in Triton Induced Hyperlipidemia
The data in Table 1 shows that acute administration of Triton WR-1339 in rats caused marked increase in their plasma levels of TC (168 %), PL (187 %) and TG (176 %) following inhibition of PHLA by 31 %. Treatment with C. tora seeds extract at the dose of 500 mg/kg, b.w., caused decrease in these levels of TC, PL and TG by 28, 25 and 26 %, respectively, simultaneously with reactivation of PHLA by 22 %. However lipid lowering action of guggulipid even at a lower dose of 200 mg/kg, b.w., was comparatively higher (33–40 %) to that of C. tora seeds extract.
Table 1
Effect of Cassia tora seeds extract and guggulipid on plasma lipids in triton induced hyperlipidemia in rats
| Groups | Total cholesterol (mg/dl) | Phospholipid (mg/dl) | Triglyceride (mg/dl) | Post heparin lipolytic activity (n mole FFA released/h/L) |
|---|---|---|---|---|
| Control | 87.52 ± 6.11 | 82.46 ± 3.33 | 90.66 ± 8.69 | 16.86 ± 0.95 |
| Triton treated | 234.60*** ± 12.09 (+168) | 235.89*** ± 12.09 (+186) | 250.57*** ± 24.86 (+176) | 11.56** ± 1.34 (−31) |
| Triton + Cassia tora (500 mg/kg, b.w.) | 169.87** ± 6.14 (−28) | 177.89** ± 8.69 (−25) | 185.88** ± 12.80 (−26) | 14.11* ± 0.70 (+22) |
| Triton + Guggulipid (200 mg/kg, b.w.) | 153.13*** ± 8.65 (−35) | 159.14*** ± 7.85 (−33) | 149.14*** ± 8.23 (−40) | 14.36* ± 0.68 (+24) |
Values are mean ± SD of 6 animals. Values in parenthesis indicate percent change. Triton treated group is compared with control, triton and drug treated with triton
* P < 0.05; ** P < 0.01; *** P < 0.001
Effect of C. tora Seeds Extract in Cholesterol Rich-HFD Induced Hyperlipidemia
In this model of hyperlipidemia (Table (Table2),2), feeding with cholesterol rich-HFD in rats caused significant increase in their plasma levels of TC (135 %), PL (72 %) and TG (119 %) following inhibition of PHLA by 32 %. Treatment with C. tora seeds extract for 15 days, reversed these plasma levels of TC, PL and TG by 25, 21 and 31 %, respectively, simultaneously with reactivation of PHLA by 21. %. The hypolipedemic action of C. tora at the dose of 500 mg/kg, b.w., was comparable to that of guggulipid at the dose of 200 mg/kg, b.w. in above model.
Table 2
Effect of Cassia tora seeds extract and guggulipid on plasma lipids in cholesterol rich-HFD induced hyperlipidemia in rats
| Groups | Total cholesterol (mg/dl) | Phospholipid (mg/dl) | Triglyceride (mg/dl) | Post heparin lipolytic activity (n mole FFA released/h/L) |
|---|---|---|---|---|
| Control | 88.59 ± 6.98 | 80.62 ± 4.29 | 114.80 ± 11.30 | 19.31 ± 1.30 |
| Cholesterol rich-HFD treated | 208.18*** ± 22.10 (+135) | 138.27*** ± 24.86 (+72) | 251.35*** ± 24.59 (+119) | 13.11** ± 1.18 (−32) |
| Cholesterol rich-HFD +Cassia tora (500 mg/kg, b.w.) | 155.36** ± 11.13 (−25) | 108.86* ± 6.78 (−21) | 172.88** ± 6.49 (−31) | 15.88* ± 1.78 (+21) |
| Cholesterol rich-HFD + Guggulipid (200 mg/kg, b.w.) | 153.39** ± 8.83 (−26) | 100.30** ± 8.77 (−28) | 169.92** ± 6.09 (−32) | 16.33 * ± 1.64 (−25) |
Values are mean ± SD of 6 animals. Values in the parenthesis indicate percent change. Cholesterol rich-HFD treated groups is compared with control, cholesterol rich-HFD and drug treated groups with cholesterol rich-HFD
* P < 0.05; ** P < 0.01; *** P < 0.001
Effect of C. tora Seeds Extract in Cholesterol Rich-HFD Induced Steosis in Liver
Feeding with cholesterol rich-HFD in rats also caused accumulation of TC (53 %), PL (72 %) and TG (57 %) following diminution of LPL activity by 37 % in their liver (Table 3). However, treatment with C. tora seeds extract exerted a decrease in these levels of TC, PL and TG by 26, 22 and 29 %, respectively, following reactivation of LPL activity (26 %) in hyperlipidemic animals. Guggulipid was more effective hypolipedemic than C. tora seeds, as it could decrease the level of lipids by 26–40 %, following reactivation of LPL activity (29 %) in the liver of hyperlipidemic rats.
Table 3
Effect of Cassia tora seeds extract and guggulipid on Liver lipid in cholesterol rich-HFD induced hyperlipidemia in rats
| Groups | Total cholesterol (mg/dl) | Phospholipid (mg/dl) | Triglyceride (mg/dl) | Lipoprotein lipase activity (n mole FFA released/h/mg protein) |
|---|---|---|---|---|
| Control | 6.75 ± 0.30 | 22.43 ± 2.71 | 4.54 ± 0.38 | 79.85 ± 4.82 |
| Cholesterol rich-HFD treated | 10.33*** ± 0.62 (+55) | 38.58*** ± 5.33 (+72) | 8.56*** ± 1.02 (+87) | 50.26*** ± 2.72 (−37) |
| Cholesterol rich-HFD + Cassia tora (500 mg/kg, b.w.) | 7.67** ± 0.62 (−26) | 29.40** ± 3.60 (−24) | 5.22*** ± 0.74 (−39) | 63.09* ± 3.75 (+25) |
| Cholesterol rich-HFD +Guggulipid (200 mg/kg, b.w.) | 7.36** ± 0.90 (−29) | 28.47** ± 5.04 (−26) | 5.11*** ± 0.62 (−40) | 64.99** ± 4.66 (−29) |
Values are mean ± SD of 6 animals. Values in the parenthesis indicate percent change. Cholesterol rich-HFD treated groups is compared with control, cholesterol rich-HFD and drug treated groups with cholesterol rich-HFD
* P < 0.05; ** P < 0.01; *** P < 0.001
Discussion
Triton WR-1339 acts as a surfactant and suppresses the action of lipases to block the uptake of lipoproteins from circulation by extra hepatic tissues, resulting in increased blood lipid concentration [16]. The lipid-lowering effect caused by feeding with C. tora seed extract, as in the case of guggulipid, may be due to an early clearance of lipids from circulation in triton model and it may be due to reactivation of lipolytic enzymes as evidenced by increased plasma PHLA [17]. We have successfully used this model for the evaluation of the lipid-lowering activity of some natural products [18, 19]. The present investigation with cholesterol rich-HFD fed hyperlipidemic animals showed that C. tora seed extract could stimulate PHLA and hepatic LPL activity, both of which play a key role in lipid catabolism and their utilization in body [20]. This situation imposed by feeding with the test sample, may be responsible for the decrease in the level of plasma and liver lipids in this model. We have reported that hypolipidemic action of guggulsterone, the active principle of guggulipid, is mediated through activation of PHLA, LPL, and lecithin cholesterol acyl transferase activities, inhibition of hepatic cholesterol biosynthesis, and increased faecal bile acid excretion [9]. The same mechanisms may also interplay to cause the hypolipidemic effect of C. tora seed extract.
Earlier studies have shown that feeding with C. tora seed extract and fibers isolated from seeds caused lowering in blood lipid levels in rats fed with high cholesterol diet [21, 22]. Chandan et al. [23], in a phyto-pharmacological overview on C. tora documented a variety of beneficial effects of the phytochemicals isolated from seed, leaf and root of this plant. Some of the main constituents of C. tora seed are anthraquinones, chrysophanol, emodin, rhein, euphol, basseol [24]. It also contains phenolic glycosides namely: rubrofusarine triglucoside, nor-rubrofusarin gentiobioside, demethyl flavasperone gentiobioside, torochrysone gentiobioside, torachrysone tetra- glucoside and torachrysone apioglucoside [2]. Seed oil contains different percentage of oleic, linoleic, palmitic, stearic and lignoceric acids [25]. It is suggested that all or some of these bioactive compounds may be responsible for hypolipidemic activity of C. tora seed. Researchers found that chrysophanic acid –9-anthrone, alateranin and rubrofusarin gentiobioside, naphthalenes and anthraquinones, naphthopyrone glucosides, ononitol monohydrate possess antifungal [26], antimutagenic [27], antibacterial [28], anti glycation [29] and hepatoprotective activities [30]. Recently Chaurasia et al. [3] isolated a napthopyrone glycoside as active antidiabetic constituent form n-butanol extract of C. tora seeds.
Here we have tested crude extract of C. tora seeds which however, upon research and development, may produce a more potent lipid lowering natural product or a pure compound like guggulipid/guggulsterone from commephora mukul. Further work on drug metabolism and to assess the biological activity in vivo and in vitro of C. tora seeds and its fraction is in progress to substantiate the present findings.
Acknowledgments
One of us (V.K.) is grateful to the Director, Central Drug Research Institute, Lucknow for experimental support and Era’s Lucknow Medical College & Hospital, Lucknow for financial support.
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