ANTIHYPERGLYCEMIC ACTIVITY OF IMPERATA CYLINDRICA ROOT METHANOL EXTRACT IN RATS

Objective: This study aims to evaluate the inhibitory activity of Imperatacylindrica root methanol extract on glucose absorption level in intestine of male Wistar strain rats at a dose of 90 mg/kg- Methods: Extract inhibitory activity on the glucose absorption level in the intestine of the rats followed Soedigdo-Marsongkohadi method. Glucose absorption level was measured from a mixture of 30 mmol glucose in 0.9 % sodium chloride solution as the control solution and a mixture of the extract at the dose of bw, which is a dose having antihypertensive activity. 90 mg/kg- Results: The result showed that the extract at the dose of bw in another control solution as the tested solution 90 mg/kg- Conclusion: I. cylindrica root methanol extract at the dose of bw had significant activity (p<0.05) to decrease the glucose absorption level in the intestine of the rats. cylindrical bw had a significant activity to inhibit the glucose absorption level in the intestine of the rats. the at the dose of 90 mg/kg-bw antihyperglycemic

The polyphenol compounds in methanol extract of I. cylindrica root antioxidant activity, which wasIC50 of 0.32 mg/ml along with the ability to donate a hydrogen atom to 1,1-diphenyl-2-pikrilhidrazil (DPPH) free radical, a total polyphenol content of 1.53% gallic acid equivalent (GAE) [6], and a total flavonoid content of1.17% quercetin equivalent (QE) [7]. Isoeugenin (new chromone) in methanol extract of I. cylindrical root as a nitric oxide synthase inhibitor is known to have a potential treatment for in flammatory disease [5], and it is also used for liver disease in ethnomedicinal plant [8]. The methanol extract is regarded as antianthelmintic at a concentration of 80 mg/ml and can cause paralysis in 3.3 min and death in 6.0 min. The potent activity of antianthelmintic is indicated by the presence of various phytoconstituents such as tannins and saponins [9]. The methanol extract at the doses of 100, 200 and 300 mg/kg, p. o had anticoagulant activity in rats [10]. The aqueous extract of I. cylindrical Indonesian people still use traditional medicine; one of the four types of traditional medicine is jamufrom herbs. Jamu is considered as natural-based medicines, which can be processed as herbmedicines [13].
aerial part has very potent antibacterial activity comparatively with the ether and ethanol extracts when tested against Staphylococcus aureus and Escherichia coli [11]. However, the root extract which is different from in the extraction method, solvent and dose does not have an effect to S. aureus (ATCC 6538P), Bacillus subtilis (ATCC 6633), Pseudomonas aeruginosa (ATCC 9027), Mycobacterium smegmatis (ATCC 14468), Candida albicans (ATCC 2091) [12].
Double burden of communicable and noncommunicable diseases increase in Indonesia and become main attention for the government. Hypertension, diabetes mellitus, cancer, and chronic obstructive pulmonary diseases are primary non-communicable disease-causing worldwide mortality, enhance the risk of strokes, heart attacks, and kidney failure, which become serious problems globally The data reported by the WHO in 2017, eighty-six diseases of most causing deaths were discovered in Indonesia. Among those diseases, stroke is regarded as the major cause of deaths. Meanwhile, coronary heart disease (CHD), diabetes mellitus (DM), and hypertension were 2 . World Health Organization (WHO) classified the main types of noncommunicable diseases (NCDs); they are cardiovascular diseases (such as heart attacks and stroke), cancers, chronic respiratory diseases (such as chronic obstructive pulmonary disease and asthma) and diabetes [14][15][16]

Experimental procedures
The protocol of the study was approved by the Animal Ethics Committee of Faculty of Medicine, Universitas Kristen Maranatha-R. S. Immanuel on No. 193/KEP/IV/2018 on 7 April 2018. Perfusion method used was a method described by Soedigdo-Marsongkohadi [18]. Before the examinations, all Wistar strain rats were placed in the laboratory for seven days. The rats were occupied in a suitable environment with several provided facilities such as normal temperature of the living room, a 12-hour of darkness/12-hour of light cycle, and food and water.
KTM-100 injection was injected via an intramuscular (IM) route to anaesthetize the rats at the dose of 0.4 ml/200 g-bw. After the rats under anesthetized state, the Then, the inhibitory activity measurement of the extract on glucose absorption considered as the tested solution. The tested solution was circulated again in the intestine of the rats and the glucose level in the tested solution was measured every 15 min for one hour. Unabsorbed glucose level was measured by spectrophotometer at wavelength 505 nm while absorbed glucose level was calculated from the unabsorbed glucose level by using the equation: the absorbed glucose levels = (100-unabsorbed glucose level) (%).
abdomen of the rats was opened and the perfusion instrument was put in the intestine where both intestine in abdomen and perfusion instrument were linked by two bent pipettes. The first pipette used was 10 cm placed from the pylorus, and the second pipette was 25 cm placed from the first pipette. Then the intestine was cleaned by 0.9% sodium chloride solution. Afterward, the control solution was circulated in the intestine four times per minute. The glucose level in the control solution was measured every 15 min for one hour. After one hour, the control solution was thrown out from the intestine; therefore, a repeated clean of the intestine was performed by 0.9% sodium chloride solution.

RESULTS AND DISCUSSION
The glucose absorption level from glucose control and a mixture of glucose and I. cylindrical root methanol extract at the dose 90 mg/kgbw (as glucose tested solution) by the intestine of the rats was measured in 15, 30, 45 and 60 min has shown in table 1 and table 2.  Breakdown of food in t %) were statistically significant (p<0.05).
he digestive system involves ingestion, digestions, then food absorption across the gastrointestinal epithelium prior to circulatory vessels occurs. In the gastrointestinal tract, nutrients are digested and net nutriome (is a term referring to all beneficial food components) is produced. Nutriome will be absorbed in the gastrointestinal tract and the main absorptive tissue in the gastrointestinal tract is the small intestine [19]. Based on the data in table 1 and 2, the study showed that the glucose absorption occurred in the intestine by using I. cylindrica root methanol extract at the dose of 90 mg/kg-bw(48.71±5.87%). The result of glucose absorption level by the intestine of the rats determined glucose absorption level of control solution was 60.28±3.33% and the glucose absorption level of the tested solution was 48.71±5.87% and student T-test between differences of glucose control and glucose tested showed significant results (p<0.05). This finding reveals The effect of chronic high glucose concentrations causes injuries of a large number of organs and tissues. As various reactions occurring inside the I. cylindrical root methanol extract at the dose of 90 mg/kg-bw has inhibitory activity on glucose absorption in the intestine. cells, in this worse condition, most of the cells can adjust the rate of intracellular glucose transport and can eliminate negative effects to the intracellular environment. Other cells which are susceptible to this effect include endothelial, neuronal, and β-cells [20]. Chronic hyperglycemia is a hallmark of diabetes mellitus and becomes a main risk factor for endothelial dysfunction and vascular complication. Vascular complications exhibit hyperinsulinemia due to compensation for insulin resistance associated with atherogenic risk (e. g., dyslipidemia, hypertension, procoagulant state) [20,21]. Endothelial cells release diverse mediators which affect the vascular tone (the balance of blood vessel as follows: vasoconstrictors such as endothelin-1, vasodilators such as nitric oxide), cell adhesion, and the homeostasis of clotting; and fibrinolysis [20][21][22][23]. Hyperglycemia disturbs cellular homeostasis and biochemistry, causing modifications both in small blood vessels (microvasculature) and large vessels (macro vasculature). Loss of normal function of ECs is marked by the following features: decreased endothelium-mediated vasorelaxation, deficiency in bioavailable nitric oxide (NO), deregulation of hemodynamic, and among others [20][21][22][23]. Vascular relaxation mechanism contributed by NO; the NO is generally produced by endothelial nitric oxide synthase (eNOS) towards vascular smooth muscle cells (SMC) to excite soluble sGC (soluble guanylyl cyclase) and producing the formation of cyclic guanosine monophosphate (cGMP). cGMP will activate protein kinase G (PKG), PKG can cause of vascular relaxation [21,22]. Chronic hyperglycemia can cause diabetes mellitus (DM). Hyperglycemia in DM may develop vascular complications such as dyslipidemia, hypertension, retinopathy, neuropathy, and nephropathy, and others [20][21][22][23]. Severe impacts of chronic hyperglycemia have been described previously in this study. Therefore, the inhibitory activity on the glucose absorption in the intestine by using I. cylindrica root methanol extract at the dose of 90 mg/kg-bwis useful to maintain health. The extract at the dose of 90 mg/kg-bw can decrease the postprandial blood glucose level. The antihyperglycemic activity can prevent the development of other severe diseases or complications with severe diseases in DM such as hypertension. Besides, the extract also has anti-hypertensive activity on hypertension rat model; CONCLUSION thereby, of I. cylindrica root methanol extract at the dose of 90 mg/kg-bw has two synergistic activities as anti-hyperglycemic and anti-hypertensive.
The I. cylindrical root methanol extract at the dose of 90 mg/kg-bw has been proved to have inhibitory activity on glucose absorption in the intestine of the rats. At the dose of 90 mg/kg-bw (the antihypertensive dose) of I. cylindrica root methanol extract have ACKNOWLEDGMENT significant activity (p<0.05) to inhibit the glucose absorption level in the intestine of the rats. Thereby, I. cylindrica root methanol extract at the dose of 90 mg/kg-bw has antihyperglycemic activity.
We want to say thanks to Halu Oleo University, who give us supports and facilities for this research.