#TITLE_ALTERNATIVE#

• Main Author: FITRIANDA NIM : 30713011, EKA
• Format: Dissertations
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/26868
• Institution: Institut Teknologi Bandung

Diabetes mellitus is a group of diseases characterized by state of hyperglycemia due <br /> <br /> to insufficient secretion of insulin, insulin action, or both. The state of chronic <br /> <br /> hyperglycemia in diabetes mellitus can lead to long-term damage, dysfunction and <br /> <br /> failure of a number of organs, especially eyes, kidneys, nerves, heart, and blood <br /> <br /> vessels. <br /> <br /> Extract of Centella asiatica or gotu kola has been shown to have antidiabetic <br /> <br /> activity both empirically and experimentally. However, investigation is needed to <br /> <br /> determine which compounds are actually responsible for the activity. Further, the <br /> <br /> action mechanism of the compounds in reducing blood glucose levels, and the <br /> <br /> safety profile of its use as a drug ingredient should also be investigated. <br /> <br /> This study aimed to find the active antidiabetic fraction and compound in leaves of <br /> <br /> gotu kola, and to find the mechanism of action of the compound, as well as to obtain <br /> <br /> safety profile of active fraction. To achieve these objectives, the research series <br /> <br /> were designed as follows: separation of saponin-rich and non saponin fractions <br /> <br /> from ethanolic extract of gotu kola leaves; investigating in vitro antidiabetic activity <br /> <br /> of extract, fractions and asiaticoside compound using alpha glucosidase inhibition <br /> <br /> method, alpha amylase inhibition method and glycation inhibition method; <br /> <br /> investigating in vivo antidiabetic activity using oral glucose tolerance test, alloxan <br /> <br /> induction method and high lipid-carbohydrate diet induction method in animal (in <br /> <br /> vivo testings were done using dosage of 125, 250 and 500 mg/kg for extract; 75, <br /> <br /> 150 and 300 mg/kg for fractions; 1.4, 2.8 and 5.6 mg/kg for asiaticocide); as well <br /> <br /> as oral acute and repeated doses toxicity of active antidiabetic fraction. <br /> <br /> In vitro activity test showed that the extract, saponin-rich fraction and asiaticoside <br /> <br /> compound of pegagan leaves were active as antidiabetes. The IC50 of extract, <br /> <br /> saponin-rich fraction and asiaticoside in inhibiting alpha glucosidase activity were <br /> <br /> 372,5, 175,5 and 14,6 &#956;g/ml, while IC50 in inhibiting alpha amylase activity were <br /> <br /> 266,8,173, 8 and 30.2 &#956;g/ml respectively. But no inhibition was detected in antiglycation test of these samples. <br /> <br /> On oral glucose tolerance test, extract, saponin-rich fraction and asiaticoside <br /> <br /> significantly suppressed blood glucose levels increase from 15 to 60 minutes after <br /> <br /> the glucose loading. The respective increased in glucose levels in the 15 <br /> <br /> th <br /> <br /> , 30 <br /> <br /> th <br /> <br /> and <br /> <br /> 60 <br /> <br /> th <br /> <br /> minutes after glucose loading in the extract group at dose of 500 mg/kg were <br /> <br /> 68%, 57% and 8%; in saponin-rich fraction at dose of 300 mg/kg were 94%, 92% , <br /> <br /> and 11%; and in the asiaticoside group at dose of 5.6 mg/kg were 92%, 79% and <br /> <br /> 7% of baseline levels (before glucose loading) respectively. The increase in glucose <br /> <br /> levels of control group were 169%, 124% and 65% of baseline. <br /> <br /> In alloxan-induced diabetic animals, extract, saponin-rich fraction and asiaticoside <br /> <br /> lowered glucose and HbA1c levels significantly and increased insulin levels, even <br /> <br /> thought there was no improvement in pancreatic Langerhans island damage due to <br /> <br /> alloxan. Significant decrease in glucose levels began after 14 days of administration <br /> <br /> of all test doses. The highest decrease of fasting blood glucose level after 21 days <br /> <br /> of administration of test preparation was shown by extract group at dose of 500 <br /> <br /> mg/kg by 21%, saponin-rich fraction at dose of 300 mg/kg by 9% and asiaticoside <br /> <br /> at dose of 5.6 mg/kg by 23%. Significant decrease in HbA1c levels compared to <br /> <br /> control group only occurred in group of extract at dose of 500 mg/kg, saponin-rich <br /> <br /> fraction at dose of 300 mg/kg, and asiaticoside at dose of 5.6 mg/kg which were <br /> <br /> 2.04, 2.01, and 2.11 ng/ml respectively, compared to HbA1c levels in the control <br /> <br /> group of 2.76 ng/ml. Significant increase in insulin levels were detected in the <br /> <br /> extract, saponin-rich fraction and asiaticoside groups at all doses. The highest <br /> <br /> insulin levels were obtained from the extract group at dose of 500 mg/kg (19.4 <br /> &#956;IU/ml), saponin-rich fraction at dose of 300 mg/kg (24.8 &#956;IU/ml), and asiaticoside <br /> <br /> at dose of 5.6 mg/kg (27.8 &#956;IU/ml), compared to the control group of 7.28 &#956;IU/ml. <br /> <br /> Meanwhile, no increase in the number of the island Langerhans and changes in the <br /> <br /> shape and size after treatment with the test materials. <br /> <br /> In insulin-resistant animal models induced with high fat-carbohydrate diet, <br /> <br /> decreased levels of glucose and HbA1c, and increased values of insulin tolerance <br /> <br /> test constant were only seen in extract and saponin-rich fraction groups, while <br /> <br /> asiaticoside did not exhibit similar activity although it could reduce fatty deposits <br /> <br /> arround the hepatocytes in test animals. Significant decrease in glucose levels <br /> <br /> started from 7 <br /> <br /> th <br /> <br /> day of dosage administration. The highest decrease of blood glucose <br /> <br /> level were measured on 21 <br /> <br /> st <br /> <br /> day in group of extract at dose of 500 mg/kg by 26%, <br /> <br /> and saponin-rich fraction at dose of 300 mg/kg by 25%. After 21 days of <br /> <br /> administration there was also a significant increase in insulin tolerance test constant <br /> <br /> values in the extract and saponin-rich fraction group. The increase of insulin <br /> <br /> tolerance test constant in extract group at dose of 500 mg/kg and saponin-rich <br /> <br /> fraction at dose of 300 mg/kg were 25% and 24% respectively. Significant decrease <br /> <br /> in HbA1c levels also occurred after 21 days of extract and saponin-rich fraction <br /> <br /> administration at all doses. Lower HbA1c levels were shown by extract at dose of <br /> <br /> 250 mg/kg (1.82 ng/ml) and saponin-rich fraction at dose of 150 mg/kg (1.79 <br /> <br /> ng/ml), compared to HbA1c in control group (2.25 ng/ml). Meanwhile, histologic <br /> <br /> observation showed that administration of extract, saponin-rich fraction, and <br /> <br /> asiaticoside reduce fat deposit in hepatocytes of liver tissue. <br /> <br /> Oral acute toxicity test results of saponin-rich fraction showed that LD50 of the <br /> <br /> fraction was higher than 5000 mg/kg. The results of oral subchronic toxicity test <br /> <br /> using repeated dose method for 28 days showed that in general saponin-rich fraction <br /> <br /> did not cause significant changes in the parameters observed. Despite changes in <br /> <br /> some hematological parameters in female animals, which showed mean corpuscular <br /> <br /> hemoglobin concentration (MCHC) values at dose of 500, 1000 mg/kg and control <br /> <br /> satellites at 22, 20, and 32 g/dl compared to control group values of 29 g/dl, and <br /> <br /> decreased platelet counts at dose of 500 and 1000 mg/kg to 310,000 and 289,000 <br /> <br /> compared to control group values of 435,000, but the values of these parameters <br /> <br /> were within normal limits. <br /> <br /> The results of in vitro and in vivo studies showed that the ethanolic extract of gotu <br /> <br /> kola leaves had antidiabetic activity by inhibiting alpha amylase and alpha <br /> <br /> glucosidase enzymes, increasing insulin levels, as well as increasing sensitivity to <br /> <br /> insulin. Saponin-rich fraction is an active antidiabetic fraction of the leaves of gotu <br /> <br /> kola, and asiaticoside compound is one of the compounds responsible for the <br /> <br /> antidiabetic activity. The dominant mechanism of action of the saponin-rich <br /> <br /> fraction as antidiabetic is increasing insulin sensitivity, beside increasing insulin <br /> <br /> levels and inhibiting the activity of alpha amylase and alpha glucosidase enzymes, <br /> <br /> while the dominant mechanism of action of asiaticoside is inhibiting the activity of <br /> <br /> alpha amylase enzyme. In addition the latter also inhibits the activity of alpha <br /> <br /> glucosidase enzyme and increase levels of insulin. The saponin-rich fraction <br /> <br /> belongs to the category of practically non-toxic substances with LD50 of higher than <br /> <br /> 5000 mg/kg and does not produce in toxicity symptoms in repeated oral dosing for <br /> <br /> 28 days. <br /> <br />