ACORUS CALAMUS L ON TYPE 2 DIABETES MELLITUS MEDICATION

Diabetes is one of the metabolic diseases indicated by hyperglycemia resulting from production in insulin secretion, insulin action, or both. Type 2 diabetes, which accounts are ~90–95% of those with diabetes, the cause is a combination of resistance to insulin action and an inadequate compensatory insulin secretory response. Adequate glycemic control is thus one of the key factors to treat and/or reduce the diabetes and many plants have been used to reduce the glucose level by inhibiting the α-glucosidase that breaks down starch and oligosaccharide to glucose. Acorus calamus L (AC) had been used in the folk medicine to treat diabetes. In vitro α-glucosidase assay was carried out by measuring the release of pnitro phenol, the insulin sensitizing activity, AC altogether brought down fasting serum glucose, and smothered the rebellion of blood glucose levels after 2g/kg glucose stacking in ordinary mice, in silico study showed that chemical compounds on AC can inhibit α-glucosidase and the later investigate is demonstrated to decide the impacts and atomic instruments of AC on glucagon-like peptide-1 (GLP-1) expression and discharge related to its hypoglycemic impacts.


INTRODUCTION
Diabetes mellitus could be a illnesses of disarranged digestion system, influenced by a combination of innate and natural components, coming about in hyperglycemia due to abandons in either affront discharge or affront activity within the body. Constant hyperglycemia amid diabetes causes glycation of body proteins that in turn leads to secondary complications affecting eyes, kidneys, nerves and courses 1 . Diabetes mellitus (DM) could be a common endocrine framework malady that causes metabolic disarranges and which leads to numerous organ harm disorder. Clinical chief naval officer diabetes is partitioned into two sorts, with more than 90% of patients having Sort II diabetes 2 . The number of diabetes cases was 171 million in 2000 and is anticipated to rise to 366 million in 2030 3 . Acting as a key chemical for carbohydrate assimilation, intestinal α-glucosidase could be a glucosidase found at the epithelium of the little digestive tract. αglucosidase has been recognized as a helpful target for the balance of postprandial hyperglycemia, which is the earliest metabolic abnormality that occurs in Type 2 DM 4 . Type 2 Diabetes mellitus (DM) Type 2 diabetes is the for the most part type of diabetes, representing around 90-95% of every diabetic case. This type of diabetes typically starts with insulin lack of care, a condition where muscle, liver and fat cells don't react to insulin appropriately. The pancreas in the end loses the capacity to deliver and discharge enough insulin in light of food consumption. Gestational diabetes is brought about by hormonal changes during pregnancy or by insulin inadequacy. The glucose neglects to enter cells, in this manner expanding the glucose level in the blood. High blood glucose, otherwise called hyperglycemia, can harm nerves and veins, prompting Complex illnesses, for example, coronary illness, stroke, kidney brokenness, visual impairment, nerve issues, gum contaminations and removal 5 . Type 2 diabetes mellitus could be a complex, multi factorial infection. Oxidative stretch has been recommended to be a contributory calculate in advancement and complication of diabetes 6,7 . In later a long time, characteristic cancer prevention agents are utilized in dietary, pharmaceutical and corrective to supplant engineered cancer prevention agents 8 . Inquire comes about finding that a few antioxidant substances separated and distinguished from restorative plants had great comes about on auto oxidation in vitro and in vivo 9, 10 . Postprandial hyperglycemia is the foremost vital wellbeing issue within the 21 st century. α-Glucosidase inhibitors decreases postprandial blood glucose level. Looking for strong normal substances as glycosidase inhibitors is exceptionally vital to remedy diabetes 11 . Diagnostic criteria for diabetes The blood glucose levels of a sound man are 80 mg/dl on fasting and up to 160 mg/dl in the postprandial state. Diabetes mellitus is described by intermittent or steady hyperglycemia, and is analyzed by exhibiting one of the accompanying: Fasting blood glucose level at or upper 126 mg/dl or 7.0 mmol/l. plasma glucose at or upper 200 mg/dl or 11.1 mmol/l two hours after a 75 g oral glucose load in a glucose resistance test. Plasma glucose at or over 200 mg/dl or 11.1 mmol/l. Two fasting glucose estimations over 126 mg/dl or 7.0 mmol/l or arbitrary glucose level >200mg/dl on two events is viewed as indicative for diabetes mellitus. Patients with fasting sugars somewhere in the range of 6.1 and 7.0 mmol/l (110 and 125 mg/dl) are considered to have debilitated fasting glucose and patients with plasma glucose at or over 140 mg/dl or 7.8 mmol/l two hours after a 75 g oral glucose load is considered to have weakened glucose resilience 12 .

Type 2 diabetes mellitus medication
After over than ten a long time of inquire about unraveling complex metabolic control systems, solutions able of a secure inversion of sort 2 diabetes are still not accessible. Verifiably, complex illnesses have over and over demonstrated to be insubordinate to the finest mono-therapeutic approaches 13 . A few illustrations of combination treatments have generally overcome such challenges, outstandingly for the treatment of extreme hypertension and tuberculosis. Corpulence and its results, such as sort 2 diabetes, have demonstrated to be similarly safe to helpful approaches based on single solutions. Suitable administration of sort 2 diabetes regularly requires adjunctive medications, and the later enrollment of some compound blends has set the point of reference for combinatorial treatment of weight. On the other hand, twofold or triple helpful combinations are more troublesome to development to administrative endorsement.
Taking after moved forward understanding of the atomic premise for metabolic benefits taking after bariatric surgery interventions, several classes of novel uni-molecular or independent combination therapeutics were discovered. These unused classes of medicate candidates are based on gastrointestinal hormones, offer adequacy predominant to right now endorsed alternatives and appear to have potential to completely switch human weight and sort 2 diabetes 14, 15 . Besides, intestine peptide-based cellspecific focused on conveyance of little atoms offers extra potential for novel metabolic exactness drugs and decreased systemic side impacts. In this introduction the revelation, pre-clinical approval and to begin with clinical tests of peptide hormone poly-agonist medicate candidates as well as of combinatorial single particle helpful candidates will be summarized, counting already unpublished perceptions 16 .

Mechanism of antidiabetic therapy
Western diabetic drugs adjust hypoglycemia by supplementing affront, moving forward affront affectability, expanding affront emission from the pancreas and/or glucose take-up by tissue cells. Beneath ordinary conditions, pancreatic β-cells discharge adequate affront to preserve blood glucose concentration inside a limit extend (72-126 mg/dl). The affront incitement taken after by cascade signaling improves glucose admissions, utilization and capacity in different tissues. In diabetic patients, the body loses affront creating capacity as a result of pancreatic β-cell apoptosis or affront lack of care. The cytokines, lipotoxicity and gluco-toxicity are three major jolts for βcell apoptosis 15 . The medicines of diabetes incorporate slim down, work out, utilize of verbal hypoglycemic specialists and affront are the essential shapes of treatment for diabetes. Right now accessible engineered antidiabetic operators other than being costly deliver genuine side impacts. Home grown pharmaceutical is separated from as of now accessible restorative choices. There are numerous home grown solutions have been suggested for the treatment of diabetes mellitus. Utilizing of restorative plants has the advantage of having no side impacts 13 . Conventional plant medicines have been utilized all through the world for the treatment of diabetes mellitus. History appeared that therapeutic plants have been utilized in conventional mending around the world for a long time to treat diabetes; this is often since such home grown plants have hypoglycemic properties and other advantageous properties, as detailed in logical literary works 14 .

Traditional plant for antidiabetic
Ethno pharmacological overviews demonstrate that more than 1200 plants are utilized in conventional medication for their affirmed hypoglycemic action 17 . Therapeutic plants, since times immemorial, have been utilized in essentially all societies as a source of pharmaceutical. A consider of old writing shows that diabetes was reasonably well known and wellconceived as a substance in old India. The information of the framework of diabetes mellitus, as the history uncovers, existed with the Indians since ancient age. Its most punctual reference (1000 BC in the Ayurveda writing) is found in legendary shape where it is said to have begun by eating Havisha 2, 18 . The NAPRALERT database records over 1200 species of plants speaking to 725 genera in 183 families expanding from the marine green growth and organisms with antidiabetic movement 19 . Over half of these have been utilized ethno-pharmacologically in conventional medication as antidiabetics, and a few 50% of these conventional cures have been examined tentatively 19 . The utilization of conventional pharmaceutical and restorative plants in most creating nations, as a standardizing premise for the upkeep of great wellbeing, has been broadly watched. Moreover, an expanding dependence on the utilize of therapeutic plants within the industrialized social orders has been followed to the extraction and improvement of a few drugs and chemotherapeutics from these plants as well as from customarily utilized home grown cures 2 . Certain herbs may lower blood glucose 20,21 ; however, their test results are subject to several factors. Firstly, each herb contains thousands of components, only a few of which may be therapeutically effective 22 .
Secondly, different parts of an herb have different ingredient profiles. Moreover, different extraction methods may yield different active ingredients. Thirdly, herbal formulae containing multiple herbs may have synergistic effects 15  Study Consider from the Rhizomes of Acorus calamus L. is broadly utilized within the treatment of diabetes in conventional people pharmaceutical of America 25 and it wins in Merak, Banten, Indonesia to progress diabetes. In any case, the diabetic effects of Acorus calamus L. have not been completely studied yet 26 . Acorus calamus L Acorus calamus L (AC), also known as 'Vacha or Sweet flag', it has been a critical herb within the Ayurvedic medication and inborn therapeutic framework for over 100 a long time. AC rhizomes have been utilized as a single sedate or as a component of certain compound sedate arrangements within the Indian Ayurvedic framework of medication for psychoneurosis, a sleeping disorder, mania, epilepsy and loss of memory 27,28,29 . It is additionally utilized within the treatment of hack, fever, bronchitis, irritation, sadness and other mental clutters, tumors, hemorrhoids, skin maladies, deadness and common debility 30 , stimulant, emetic, carminative, stomachic, as cures for a few harming 29 . AC can be found developing in Central Asia or India, Central Europe and North America. In India it is common in ranges that encompass the Himalayas. Indian AC from the Jammu zone is triploid and tetraploid; and European as well as American assortment of the AC is diploid 28 Botanical description: Calamus could be a semi seagoing herb and is broadly disseminated by the edges of lakes and moderate streaming waterways, growing in shallow water or in a really damp loamy soil. It lean towards a pH within the run 5.5 to 7.5. It is perennial herb; the rhizomes commonly occur in pieces about 5 to 15 cm in length and 1 to 2 cm in thickness. They are covered with a thin brownish epidermis and cork and are much shrunken, bearing brief longitudinal wrinkle. They are marked on the upper surface with large triangular leaf scare that encircle the rhizome, springing from each side alternately; from these scars fibrous leaf trace bundle frequently project. The under surface bears an irregular zigzag line of small raised root scars that are circular and exhibit a central stele surrounded by a narrow cortex. The rhizome breaks with a short corky fracture, and is pale brown or nearly white and spongy internally. The section shows a expansive stele isolated by a yellowish line, the endodermis from a thick cortex; various little, oval, vascular bundles are scattered thought the segment. The naturally broken rhizome has a pleasing fragrant odor. Clears out are right green having sword-shaped, based equitant, thickened in center, edges wavy. Blossoms are showed up in June and July and are yellow/green in color. The blossoms are bisexual (have both male and female organs) and are pollinated by Creepy crawlies 28,29,33 .

Plant
Root Figure 1: Acorus calamus L

Inhibitory of α-glucosidase
Our previous research find the potency of fraction nbutanol AC extract as inhibitory agent on αglucosidase enzyme. Sample from fraction of n-butanol AC extract with column chromatography method to separated it. We use the resin to separate fraction because it is suitable for the crude extract with high polarity (hydrophilic). The result of inhibitory assay of α-glucosidase from fraction showed that the 5 th fraction was the most active with IC50 value 4.87 µg ML-1 while the other fraction has not activity 37 . Our examination utilize a Koji extricate as control from Apergillus terreus is an particularly productive maker of auxiliary metabolites has organic exercises such as inhibitory of α-glucosidase and it features a most potential action in this manner inspected the impact on postprandial blood glucose level after a supper in mice.

In silico study
The large scale atom of the protein α-glucosidase was gotten through the protein information bank with the code 1lwj within the download NCBI site. Models of chemical compounds contained in A. calamus L. gotten through the location take out from databases "jamu" Knapsack 42 and make the 2D and 3D utilizing by Chemsketch on the freeware adaptation. At that point docking utilized Argus lab. Docking results as shown on Table 1, and Figure 3 and Figure 4. Recently, In silico has lead an important role in drug design and finding the drugs substances. In which micro molecule is virtually docked in to a drug target and the binding affinities are estimated using simplified free energy calculation method. Many programs capable of carrying out virtual screening have been developed; most of them are pay ware.  Isocaesp-itol, Acoragermacrone, Preisocalamendiol, Shyobunon, Epishyobunone, Isocalamone, Acolamone, Aristolene, (-)-Cadala-1,4,9-triene, Isocalamendiol, Calacone, β-gualene, Calamusenon, Acoronene, Acorid acid, Calarene, Acorenone through the site. Take out "jamu" Knapsack and made in the formula structures of 2D and 3D using the program ACD/Chemsketch. Then docking used Argus lab Program are visualized by Pymol program. Docking results showed activity in the compound 1-ethenyl-1-methyl-2,4-at (prop-1-en-2yl) Cyclohe-xane with free energy -8.04385 kcal/mol, and the compound Isocaespitol with a free energy -8.28388 kcal/mol.

Figure 4: Isocaespitol
Chemical component that has the lowest free energy showed the most stable affinity that is expected to have good medicinal properties as well. Docking results showed activity in the compound 1-ethenyl-1-methyl-2,4-at (prop-1-en-2-yl) Cyclohexane with free energy -8.04385 kcal/mol, and the compound Isocaespitol with a free energy -8.28388 kcal/mol 43 . Molecular mechanisms on Glucagon-like peptide-1 (GLP-1) ACE acts as an antidiabetic through affront sensitizing, affront discharging and alpha-glucosidase inhibitory exercises. The show consider is planned to examine the impacts and atomic instruments of ACE on glucagonlike peptide-1 (GLP-1) expression and secretion related to its hypoglycemic effects. The hypoglycemic effect of ACE (100mg/kg, i.g.) was affirmed by testing blood glucose levels or by means of verbal glucose resilience test (OGTT) in streptozotocin (STZ) actuated hyperglycemic mice, db/db diabetic mice and dietinduced hefty (DIO) mice. Plasma affront, GLP-1 levels and intestinal GLP-1 related quality expression were decided in STZ induced and db/db diabetic mice. The in vitro effects of ACE (12.5μg/ml) on the expression and secretion of GLP-1 were detected in NCI-H716 intestinal L-cells, and the relationship between ACE and particles within the signaling pathway was encouraging investigated 45 . ACE (100mg/kg) altogether brought down fasting blood glucose in STZ-induced and db/db diabetic mice and progressed the OGTT in DIO mice. Affront discharging and islet defensive impacts, at the side the expanded emission of GLP-1, were watched. The expression of proglucagon quality (gcg) and posttranslational processing gene prohormone convertase 3 (pc3) and the GLP-1 substance within the culture medium of L-cells eminently expanded after the ACE treatment (12.5μg/ml). At the same time, β-catenin atomic translocation happened, and its downstream protein cyclin D1 was actuated, appearing the inclusion of Wnt signaling. ACE might enact Wnt signaling to extend the quality expression of gcg and pc3 and apply incretin impacts, counting insulin otropic and islet security, to lower blood glucose levels through lifted GLP-1 discharge either straightforwardly or by implication 45 .

CONCLUSION
Acorus calamus L. has been proofed as folk medicine that can cure type 2 diabetes mellitus on many mechanisms and can be used as antidiabetic related with many experiment methods.