Diabetes remains a major health problem worldwide.

Low-risk low-cost alternatives to

pharmaceutical interventions are needed where lifestyle modifications have failed. There
has been an enormous interest in the development of alternative medicines for type 2
diabetes, specifically searching for therapeutically effective with the ability to delay or
prevent glucose absorption. The goal of the present study was to provide in vitro evidence
for potential inhibition of α-amylase enzyme. Hence α-amylase inhibition may be possible
with one of the mechanisms for the MMC extract to exert anti diabetic activity and
indicates that MMC extract can be considered as a potential candidate for the management
of type 2 diabetes mellitus.

In vitro α-amylase inhibition study

Acarbose-like drugs, that inhibit α-amylase present in the epithelium of the small
intestine, have been demonstrated to decrease post-prandial hyperglycaemia (Sima &
Chakrabarti, 2004) and improve impaired glucose metabolism without promoting insulin
secretion in NIDDM patients (Carrascosa et al., 2001). These medications are most useful for
people who have just been diagnosed with type 2 diabetes and who have blood glucose levels
only slightly above the level considered serious for diabetes. Therefore, the retardation and delay
of carbohydrate absorption with a plant-based α-amylase inhibitor offers a prospective
therapeutic approach for the management of type 2 diabetes mellitus and borderline patients
(McCue et al., 2005).

Herbal medicines with antidiabetic potential have different modes of action—

mimic insulin, act on insulin secreting beta cells, or modify glucose utilization. Herbs
which modify glucose utilization act by altering the viscosity of gastrointestinal contents,
delaying gastric emptying, or delaying glucose absorption [2]. Absorption of glucose can
be delayed by reducing the rate of digestion of starch. Inhibition of the mammalian alpha
amylase enzyme in the intestine would delay the degradation of starch and
oligosaccharides to monosaccharides before they can be absorbed. This would decrease
the absorption of glucose and consequently reduce postprandial blood glucose level [5].
Therefore, screening of alpha-amylase inhibitors in medicinal plants has received much
attention.

Materials and Methods

Collection of Plant Material
The plant material (leaves) was collected from Chennai, India in January, 2011. The plant
material was duly authenticated by Dr. Jayaraman Director at National Institute of Herbal
Science (PARC), Chennai (Voucher number: PARC/2011/742).
Preparation of Extract
The leaves were air-dried, pulverized by grinding using mortar and pestle. Thereafter, the
coarse powder of air-dried leaf
 Analysis of Primary Metabolites
The primary metabolites like carbohydrates, total proteins, and lipid contents were
quantified. Carbohydrates were quantified by the method of McCready et al. [8], proteins
by Lowry et al. [9], and lipids by Zak et al. [10].

Analysis of Secondary Metabolites

Secondary metabolites like tannins, phenols, and flavonoids were quantified in all the
individual extracts.
Estimation of Total Phenols
Estimation of Total Tannins
Estimation of Total Flavanoids
Assay of Amylase Inhibition

Results and Discussion

Hyperglycemia has been a classical risk in the development of diabetes and the
complications associated with diabetes. Therefore control of blood glucose levels is
critical in the early treatment of diabetes mellitus and reduction of macro- and
microvascular complications. One therapeutic approach is the prevention of carbohydrate
absorption after food intake, which is facilitated by inhibition of enteric enzymes
including α-glucosidase and α-amylase present in the brush borders of intestine.
In this study, the α-amylase inhibitory activity of MMC was investigated. The inhibitory
effect was analysed. The percentage inhibition of α-amylase by the extracts MMC was
studied in a concentration range of 10–640 μg/mL.
Result
The in vitro α-amylase inhibitory studies demonstrated that MMC extract had α-amylase
inhibitory activity. The percentage of inhibition at 62.69, 61.65, 57.51, 37.22 mg/ml
concentrations of MMC extract showed a concentration-dependent reduction in percentage of
inhibition. Thus the highest concentration of 100 µg /ml showed an inhibition of nearly 61.65%.
The moderate concentration of 50 µg /ml showed a maximum inhibition of nearly 62.69%. The
percentage inhibition varied from 62.69–37.22% from the highest concentration to the lowest
concentration of100 µg /ml -12.25 µg /ml.

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Background

DM (Diabetes mellitus) is a metabolic disorder characterized by chronic

hyperglycemia or increased blood glucose levels with disturbances in
carbohydrate, fat and protein metabolism resulting from absolute or relative lack
of insulin secretion [1]. The frequency of this disorder is on the rise globally, is
likely to hit 300 million by 2025 with India projected to have the largest number
of diabetic cases [2].
Type 2 diabetes is one of the primary threats to human health due to increasing
prevalence, chronic course and disabling complications [3]. Many diverse
therapeutic strategies for the treatment of Type 2 diabetes are in use. The
conventional available therapies for diabetes include stimulation of endogenous
insulin secretion, enhancement of the action of insulin at the target tissues, oral
hypoglycemic agents, such as biguanids and sulfonylureas and the inhibition of
degradation of dietary starch by glycosidases such as α-amylase and α-
glucosidase [4].
Pancreatic α-amylase is a key enzyme in the digestive system and catalyses the
initial step in hydrolysis of starch to a mixture of smaller oligosaccharides
consisting of maltose, maltotriose, and a number of α-(l-6) and α-(1 - 4)
oligoglucans. These are then acted on by α-glucosidases and further degraded to
glucose which on absorbtion enters the blood-stream. Degradation of this dietary
starch proceeds rapidly and leads to elevated post-prandial hyperglycemia. It has
been shown that activity of human pancreatic α-amylase in the small intestine
correlates to an increase in post-prandial glucose levels, the control of which is
therefore an important aspect in treatment of type 2 diabetes[5]. Hence,
retardation of starch digestion by inhibition of enzymes such as α-amylase plays a
key role in the control of diabetes. Inhibitors of pancreatic α-amylase delay
carbohydrate digestion causing a reduction in the rate of glucose absorption and
lowering the post-prandial serum glucose levels [6]. Some inhibitors currently in
clinical use are acarbose and miglitol which inhibit glycosidases such as α-
glucosidase and α-amylase while others such as and voglibose inhibit α-
glucosidase. However, many of these synthetic hypoglycemic agents have their
limitations, are non-specific, produce serious side effects and fail to elevate
diabetic complications. The main side effects of these inhibitors are
gastrointestinal viz., bloating, abdominal discomfort, diarrhea and flatulence [7].
Herbal medicines are getting more importance in the treatment of diabetes as
they are free from side effects and less expensive when compared to synthetic
hypoglycemic agents [8,9].
In India, indigenous herbal remedies such as Ayurveda and other Indian
traditional medicine have since ancient times used plants in treatment of
diabetes[10]. Ethnobotanical studies of traditional herbal remedies used for
diabetes have identified more than 1,200 species of plants with hypoglycemic
activity [3,11]. A number of medicinal plants and their formulations are used for
treating diabetes in the traditional Indian Ayurvedic system as well as in
ethnomedicinal practices. Even though, these traditional practices are empirical in
nature, over 200 million people in India with limited access to primary healthcare
centres, depend on traditional system of medicine to cater to their healthcare
needs [12]. However, this traditional knowledge, derived empirically, has to be
supported by scientific testing. WHO (World Health Organization) (1980) has
recommended the evaluation and mechanistic properties of the plants effective in
such systems [13,14]. The search for new pharmacologically active agents
obtained by screening natural sources such as medicinal plants or their extracts
can lead to potent and specific inhibitors for α-amylase [6]. Pharmacological
properties α-glucosidase inhibitors such as acarbose that can also inhibit
pancreatic α-amylase revealed that the complications of DM such as onset of
renal, retinal, lens and neurological changes and the development of ischaemic
myocardial lesions are prevented or delayed [15]. Long-term day-to-day
management of diabetes, with acarbose is well tolerated and can improve
glycaemic control as monotherapy, as well as in combination therapy [16].

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