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Cover page of the Journal of Health Sciences

 Table of Contents  
Year : 2016  |  Volume : 9  |  Issue : 3  |  Page : 308-314

Assessment of antidiabetic potential of Shilajatvadi Lauha processed with Daruharidra in streptozotocin-nicotinamide-induced diabetic rats

1 Department of Rasa Shastra and Bhaishajya Kalpana, Government Ayurved College, Raipur, Chhattisgarh, India
2 Department of Rasa Shastra, Faculty of Ayurveda, Institute of Medical Sciences, Banaras Hindu University, Varanasi, Uttar Pradesh, India
3 Department of Pharmaceutics, Neuropharmacology Research Laboratory, Indian Institute of Technology (Banaras Hindu University), Varanasi, Uttar Pradesh, India

Date of Web Publication21-Dec-2016

Correspondence Address:
Dr. Thakur Rakesh Singh
Department of Rasa Shastra and Bhaishajya Kalpana, Government Ayurved College, Raipur - 492 010, Uttar Pradesh
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/2349-5006.196334

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Objectives: To assess the antidiabetic potential of Shilajatvadi Lauha (SL) processed with Daruharidra (modified SL [MSL]) in streptozotocin (STZ)-nicotinamide (NA)-induced diabetic rats.
Materials and Methods: Animals were divided into diabetic and nondiabetic groups. Type 2 diabetes in rats was induced with a single dose of STZ (65 mg/kg) NA (110 mg/kg) intraperitoneal diabetic rats were treated with formulation MSL (10, 30, and 100 mg/kg) and glibenclamide (10 mg/kg) once daily for 14 days orally. After 14 days treatment, fasting blood glucose and plasma insulin were assayed. Different biochemical parameters such as total cholesterol (TC), triglycerides (TGs), low-density lipoprotein-cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C), and very LDL-C (VLDL-C) were also examined.
Results: MSL significantly lowered the blood glucose and increases insulin level, which was comparable to the standard antidiabetic drug, glibenclamide. Treatment with MSL showed a significantly reduction in the levels of TC, TG, LDL-C, and VLDL-C and increases the level of HDL-C.
Conclusion: MSL possess significant antidiabetic and antihyperlipidemic activity in Type 2 diabetes mellitus rats. The results are encouraging and further studies can be used to evaluate the exact mechanism of action to develop a novel molecule that will benefit the ailing.

Keywords: Antidiabetic, antihyperlipidemic, Daruharidra, herbomineral formulation, Shilajatvadi Lauha

How to cite this article:
Singh TR, Gupta LN, Kumar N, Kumar V. Assessment of antidiabetic potential of Shilajatvadi Lauha processed with Daruharidra in streptozotocin-nicotinamide-induced diabetic rats. Indian J Health Sci Biomed Res 2016;9:308-14

How to cite this URL:
Singh TR, Gupta LN, Kumar N, Kumar V. Assessment of antidiabetic potential of Shilajatvadi Lauha processed with Daruharidra in streptozotocin-nicotinamide-induced diabetic rats. Indian J Health Sci Biomed Res [serial online] 2016 [cited 2020 Jul 11];9:308-14. Available from: http://www.ijournalhs.org/text.asp?2016/9/3/308/196334

  Introduction Top

Diabetes mellitus is a chronic metabolic disorder affecting approximately 1.5% of the total population that continues to present a major worldwide health problem.[1] India leads the world with the largest number of diabetic patients earning the dubious distinction of being termed the “diabetes capital of the world.”[2] According to the International Diabetes Federation, the number of people with diabetes in India currently around 40.9 million is expected to rise to 69.9 million by 2025.[3] Diabetes mellitus is characterized by absolute or relative deficiencies in insulin secretion and/or insulin action associated with chronic hyperglycemia and disturbances of carbohydrate, lipid, and protein metabolism. As a consequence of metabolic derangement in diabetes, various complications develop, including both macro- and micro-vascular dysfunctions.[4] Despite continuing introduction of hypoglycemic drugs, diabetes and related complications still are a major global medical problem and successful treatment is of great interest but yet to be discovered.[5] The modern drugs, including insulin and other oral hypoglycemic agents such as thiazolidinediones, biguanides, and sulfonylureas, control the blood sugar level as long as they are regularly administered, but they also produced lots of undesirable effects.[6] As a result, there is a growing need for new oral antidiabetic drugs from traditional medicines as an effective and safe alternative therapy in the management of diabetes mellitus.

Rasaushadhies (herbomineral formulation) play an important role in Ayurvedic therapeutics because of their qualities such as effective atlower doses and fast acting.[7] They are prepared by addition of bhasmas (ashes)/rasayogas (mineral preparations)/ processed minerals with herbal drugs and finally they get triturated in mortar and pestle. Many herbal products, several metals, and minerals have been prescribed for the cure of diabetes mellitus alone or in combination with oral hypoglycemic agents in ancient literature.[8],[9]Shilajatvadi Lauha (SL) is one of the herbomineral formulations, which is used in the Indian System of Medicine since ancient times for treating Prameha (diabetes mellitus), Kshaya (tuberculosis), etc.[10] It contains pure Shilajit (black bitumen/mineral pitch), Swarna Makshika Bhasma (ash of chalcopyrite), Shunthi (Zingiber officinale Roscoe), Maricha (Piper nigrum L.), and Pippali (Piper longum L.) in equal quantity and Lauha bhasma (calx of iron-turning) are six times to other ingredients.[11] Antidiabetic activity of SLwas reported in nicotinamide (NA)-streptozotocin (STZ)-induced diabetic rats.[12]Daruharidra (Berberis aristata D.C. ) is an edible plant commonly used in Indian System of Medicine as a hypoglycemic, antidiarrheal, anticancer, hypotensive, central nervous system depressant, etc.[13] The root of this plant possess antihyperglycemic and antioxidant potential, hence it plays a major role in metabolic disorders.[14],[15]Daruharidra is also included in developing new herbomineral formulations such as D-400 which is used to treat diabetes mellitus.[16] By considering all these points, an attempt has been made to more potentiate SL by bhavana (levigation) with a decoction of Daruharidra root for seven times and named as Daruharidra bhavita SL/modifiedSL (MSL). In the present study, antidiabetic effect of MSL was investigated in NA-STZ-induced diabetic rats.

  Materials and Methods Top

Preparation of modified Shilajatvadi Lauha

SL was prepared by processed Shilajit, Swarna Makshik Bhasma, powders of Shunthi (Z. officinale Roscoe), Maricha (P. nigrum L.), and Pippali (P. longum L.) were taken in equal proportions in mortar and six times of Lauha bhasma was added in it and triturated with pestle until it becomes fine powder and homogenously mixed named asSL.[17] Fine powder of SL was levigated (bhavana) with a decoction of Daruharidra root [18] for continuously 6 h using a mechanized mortar and pestle with average 60 rpm. Levigation process of SL was repeated six times more with each time using fresh decoction. The final product obtained was named as Daruharidra bhavita SL/MSL.


STZ was brought from the Sisco Research Laboratories (Mumbai, Maharashtra, India). NA was provided by SD Fine Chemicals (Mumbai, Maharashtra, India). Glibenclamide was obtained from Cipla Pharmaceuticals Pvt. Ltd., India. Plasma glucose and biochemical estimation kit were procured from Span Diagnostics (Surat, Gujarat, India) and Insulin ELISA kit was obtained from Dia Metra Company (Segrate, Italy). Other chemicals were of analytical reagent grade from commercial sources.


The protocol of the study was approved by the Central Animal Ethics Committee (Dean/13-14/CAEC/327 dated November 20, 2013) for the purpose of control and supervision of experiments on animals. The experiment was carried out on healthy adult Charles Foster male Albino rats weighing 150 ± 10 g. Rats were housed in groups of six in polypropylene cages at an ambient temperature of 25 ± 1°C and 45–55% relative humidity, with a 12:12 h light/dark cycle. Animals were provided with commercial food pellets and water ad libitum. Experiments were conducted between 09:00 and 14:00 h. Principles of laboratory animal care (NIH publication number 85–23, revised 1985) guidelines were followed. Rats were equally divided in groups of six animals each except for control in which 12 rats were taken.

Drug treatments

All drugs were suspended in 0.3% w/v carboxymethylcellulose (CMC) for oral administration in animals. All doses were administered 1 h before the start of the experiments. Applications volume in all cases was 10 ml/kg, and the control animals were treated accordingly with 10 ml/kg of 0.3% w/v CMC. Graded doses of MSL were selected based on a pilot study to establish a dose-response relationship.[19]

Induction of diabetes

Type 2 diabetes was induced in overnight fasted adult male Albino Wistar rats (200–250 g) by a single intraperitoneal (i.p.) injection of 65 mg/kg STZ (dissolved in citrate buffer, pH 4.5), followed by the i.p. administration of 110 mg/kg of NA (dissolved in normal saline).[20],[21] Hyperglycemia was confirmed by elevated blood glucose levels at 72 h and then on day 7 after injection.[22] Those animals with fasting blood glucose level >200 mg/dl were considered as diabetic and were used for antidiabetic studies.

Experimental design

The rats were randomly divided into six groups each consisting six animals:

  • Group 1: Normal control (vehicle-treated)
  • Group 2: Diabetic control (vehicle-treated)
  • Group 3: Diabetic rats + MSL 10 mg/kg/day
  • Group 4: Diabetic rats + MSL 30 mg/kg/day
  • Group 5: Diabetic rats + MSL 100 mg/kg/day
  • Group 6: Diabetic rats + glibenclamide 10 mg/kg/day.

Drug treatment was started on the 7th day after induction of diabetes (day 1 of treatment). All the drugs were orally administered in the form of a suspension in 0.3% CMC, once daily for 14 consecutive days. Body weight of rats was recorded periodically. Plasma glucose was measured on the 7th (day 1 of treatment) and 14th day posttreatment. All other biochemical parameters were measured on the 14th day posttreatment.

Biochemical investigations

On day 14th after overnight fasting, blood samples were collected from the retro-orbital venous plexus under light ether anesthesia using a glass capillary tube and plasma was separated immediately. Fasting blood glucose and lipid profile were analyzed by using a biochemical kit. Plasma insulin was measured by Enzyme Linked Immunosorbent Assay kit using BIO-RAD microplate reader.[21],[23],[24]

Statistical analysis

Data of all the experiments were expressed as mean ± standard error of the mean of animals in each group (n = 6). Differences among different treatment groups were determined by one-way analysis of variance followed by Student-Newman–Keuls tests. Graph Pad Prism (version 5.03, GraphPad Software, USA) was used for statistical analysis.

  Results Top

Effect on fasting blood glucose

Fasting blood glucose levels of animals challenged with NA-STZ were significantly higher compared to the normal control animals (P < 0.001). All animals survived without any side effect and mortality during the study period. In the diabetic control group, blood glucose level remained elevated till the 14th day of the experiment. Oral administration of MSL (10, 30, 100 mg/kg/day) to diabetic rats significantly reduced fasting blood glucose compared to vehicle-treated diabetic control rats (P < 0.001) on the 14th day. On the last 14th day of the experiment, there was no statistically significant difference between the blood glucose levels of the MSL 100 mg/kg-treated diabetic group and the normal control group; such was also the case for the 10 mg/kg glibenclamide-treated diabetic group. MSL in 100 mg/kg dose almost normalizes the blood glucose level in 14 days of study [Table 1].
Table 1: Effect of modified Shilajatvadi Lauha on plasma glucose, serum insulin, and lipid profile of diabetic rats

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Effect on plasma insulin

Plasma insulin level of rats subjected to NA-STZ challenge was significantly reduced (P < 0.001) compared to normal rats. All the doses of MSL significantly increased plasma insulin level compared to diabetic control group (P < 0.001), but they did not normalize the reduced insulin level of diabetic rats in the treatment of 14 days. Glibenclamide-treated diabetic rats also show a significant increase (P < 0.001) in plasma insulin level compared to vehicle-treated diabetic control rats, and it was very much comparable to 100 mg/kg-treated MSL group [Table 1].

Effect on lipid profile

NA-STZ challenged rats (diabetic control group) after the 14th day of the study showed a significant elevation (P < 0.001) in plasma total cholesterol, triglycerides (TGs), low-density lipoprotein-cholesterol (LDL-C), and very LDL-C while the plasma high-density lipoprotein-cholesterol (HDL) level decreased significantly (P < 0.001). Although the repeated dose administration of MSL to diabetic rats significantly reversed, these changes to near normal levels (P < 0.001). The effect of MSL (100 mg/kg) was comparable with that of glibenclamide at 10 mg/kg [Table 1].

  Discussion Top

Diabetes is fast gaining the status of a potential epidemic in India with more than 62 million diabetic individuals currently diagnosed with the disease.[25],[26] It is a metabolic disorder characterized by hyperglycemia and alternation in carbohydrates, fat, and protein metabolism associated with absolute or relative deficiencies in insulin secretion or insulin action. It is considered as one of the five leading causes of death in the world.[27] Among many forms of diabetes mellitus, Type 2 occurs predominantly and affects the major population, i.e., 90% of diabetic patients. STZ selectively destroys insulin-producing pancreatic beta cells by inducing DNA-strand break in these cells through glucose transporter 2.[28] This causes activation of poly (ADP-ribose) polymerase, resulting in a reduction of cellular NAD +, and cell death. NA partially protects the beta cells against the STZ-mediated cytotoxic damage. NA was found to preserve the intracellular pool of NAD + either by acting as a precursor of NAD + or by inhibiting the activity of poly (ADP-ribose) polymerase.[29] Therefore, coadministration of NA and STZ produces stable, moderate hyperglycemia suitable for chronic diabetes study, and it produces abnormal levels of blood glucose and plasma insulin. Hence, these major parameters were taken into consideration while investigating the hypoglycemic effect of MSL.

The standard therapy for Type 2 diabetes mellitus includes diet, exercise, use of oral hypoglycemic drugs, and/or subcutaneous insulin injections.[30],[31] Several studies have demonstrated that various pharmacological agents can successfully lower blood glucose and reduce the risk of developing diabetic complications. However, a large number of limitations and unwanted side effects that still exist limit their use in clinical practice.[6],[32] Consequently, diabetic patients are also advised to support complementary and alternative therapies.

SL is one of the traditional herbomineral formulations used for the treatment of various ailments such as Prameha (diabetes mellitus) and Rajayakshma (tuberculosis).[10],[12] It contains pure Shilajit, Swarna Makshika Bhasma (ash of chalcopyrite), Shunthi (Z. officinale Roscoe), Marich (P. nigrum L.), and Pippali (P. longum L.) in equal quantity and Lauha bhasma (ash of iron-turning) are six times to other ingredients.[11],[17] Hypoglycemic effect of SL has already been proved by Singh et al.[12]Daruharidra (B. aristata) is one of the extensively used herbal medicines for treating various ailments such as diabetes, eye and ear diseases, rheumatism, jaundice, stomach disorders, skin disease, malarial fever, and as tonic.[33],[34] Several studies, mostly performed in the Chinese population, have reported the effects of berberine on the lipidic and glycemic profile.[35],[36] Several researches have also revealed the broad spectrums of therapeutically interesting bioactivities of Shilajit compound formulation.[37],[38] In Ayurvedic literature, there are many formulations which are said to be more potent if levigated with liquids (swarasa, kwatha, etc.), for example, Amalaki Rasayana and Gandhak Rasayana.[39],[40] In the present study, we examined the effect of Daruharidra levigated (bhavita) SL (MSL) in STZ-NA-induced diabetic rats. Literature search did not reveal any systematic dose finding study necessary for properly defining the therapy relevant pharmacological activity profile of MSL. Consequently, based on our pilot study to establish dose-response relationship, three graded doses, i.e., 10, 30, and 100 mg/kg/were selected for the antidiabetic study of MSL.[19]

Rats subjected to STZ-NA challenge showed significant increases plasma glucose and decreases insulin. Changes in blood glucose and insulin levels reflect abnormalities in β-cells function or structure. STZ impairs glucose oxidation and decreases insulin biosynthesis and secretion.[41],[42] Two weeks repeated oral administration of MSL and glibenclamide resulted in a significant reduction in plasma glucose level and increases insulin level of diabetic rats. It is well established that glibenclamide produces hypoglycemia by increasing the secretion of insulin from the existing pancreatic β-cells.[43] In a study, the treatment of moderate diabetic rats with some herbomineral drugs resulted in the stimulation of β-cells of  Islets of Langerhans More Details, showing an insulinotropic effect.[44],[45],[46] In the present study, though all the doses of MSL significantly lowered the fasting blood glucose and increases plasma insulin level in NA-STZ-induced diabetic rats. MSL in 100 mg/kg-treated group almost normalizes the blood glucose and insulin level and was very much comparable to glibenclamide (10 mg/kg)-treated group. These observations strongly suggest that like many therapeutically used antidiabetic drugs;[44] stimulation of insulin increase from remnant beta cells is involved in the observed effects of MSL in diabetic rats. Lipids play an important role in the pathogenesis of diabetes mellitus. Hyperlipidemia is a recognized consequence of diabetes mellitus demonstrated by the elevated levels of phospholipids, tissue cholesterol, etc. The level of serum lipids usually raised in diabetes and such an elevation represents a risk factor for coronary heart diseases.[47] The levels of cholesterol and TGs were raised in diabetic rats but which were lowered significantly with the treatment of MSL. At the same time, the level of HDL was significantly increased in the MSL-treated group. Daruharidra exerts hypolipidemic activity [48] and by repeated levigation with a decoction of Daruharidra to SL, it increases the hypolipidemic activity of MSL, and it could represent a protective mechanism against the development of atherosclerosis, which is usually associated with diabetes.

Among the various ingredients of MSL have been reported for hypoglycemic effect; Daruharidra (root bark) have important constituents like berberine, berbamine, Karachine, etc.[49],[50] Berberine has a protective effect for diabetes through increasing insulin expression, β-cell regeneration, antioxidant enzyme activity, decreasing lipid peroxidation, and HbA1C.[51] It is revealed that root of Daruharidra has a strong antihyperglycemic and antioxidant potential to regulate glucose homeostasis through decreased gluconeogenesis and oxidative stress.[15]Shilajit has pancreatotrophic action, i.e., it increases the number of β-cells of the pancreas which may result in a better sensitivity of pancreatic β-cells with the prompt secretion of a large quantity of insulin in response to hyperglycemia. It contains flavonoids and polyphenols which are natural antioxidants and significantly increases glutathione, catalase activities, and superoxide dismutase, which are the first line defensive enzymes against free radicals.[52],[53],[54]Lauha bhasma reduces the degree of oxidative stress signaling pathways and by that preventing insulin resistance and β-cells dysfunction and ultimately controlling the blood sugar level.[55]Medohara (hypolipidemic) effect of Lauha bhasma decreases the high lipid level.[56]Swarna Makshik Bhasma contain copper which is responsible for cholesterol and glucose metabolism in the body.[57],[58]Trikatu; Shunthi (Z. officinale Roscoe), Marich (P. nigrum L.), and Pippali (P. longum L.) increase bioavailability bypromoting rapid absorption from the gastrointestinaltract or preventing metabolism/oxidation during the firstpassage through the liver after being absorbed or acombination of these mechanisms helping improve most drugs therapeutic activity and it possess anti-glycation and antioxidants properties too which reduces the blood sugar level.[56],[59],[60] It has been shown that MSL markedly improved the glucose tolerance and renormalizing the serum lipid and cholesterol level in NA-STZ-induced diabetic rats, it may due to synergistic action of ingredients of MSL on peripheral uptake of insulin by the cells along with the insulinotropic action, antihyperglycemic, bioenhancer, antioxidant, rejuvenator, and free-radical scavenging activity. It has been demonstrated through a validated and systematic study that Daruharidra bhavita SL (MSL) exerts significant antidiabetic activity in rodents.

  Conclusion Top

The data of our study revealed that Daruharidra bhavita SL exerts significant antidiabetic hypoglycemic activity in NA-STZ-induced diabetic rats in a dose-dependent manner. It also showed good antihyperlipidemic property which can be attributed to the synergistic effect of multiple herbomineral ingredients of the formulation. Our results are supporting its use as a traditional medicine for the treatment of diabetes. Further studies should be undertaken to elucidate the mechanism of action of individual ingredients of Daruharidra bhavita SL, and clinical study is warranted for establishing therapeutic effectiveness.


Authors are thankful to Dr. Galib (Assistant Professor, Department of Rasashastra and Bhaishajya Kalpana, IPGT and RA, Gujarat Ayurved University, Jamnagar) and Dr. Ajit Kumar Thakur (Assistant Professor of Pharmacology, Delhi Pharmaceutical Sciences and Research University, New Delhi) for his partial support in the preparation of manuscript and experiments.

Financial support and sponsorship


Conflicts of interest

There are no conflicts of interest.

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