Potensi Protein Kecambah Kedelai dalam Menstimulasi Sekresi Insulin pada Pankreas Tikus Normal dan Diabetes

Abstract

The objective of this investigation was to examine the potential of seed and germinated SP that were extracted at two condition precipitation, Kunitz Trypsin Inhibitor (KTI) and Bowman Birk Inhibitor (BBI) to stimulate insulin secretion of the pancreas of induced diabetic and normal rats by in vitro bioassay. Mature male Sprague Dawley (SD) rats of the in vitro bioassay were divided into two groups. They were diabetic rats that was induced by aloksan injection and normal rats (without aloksan injection). Rat was anaesthetized with ether, and the pancreas was taken out, injected by HBS solution, hydrolyzed by collagenase and then washed several times with RPMI 1640 LITE solution and Kreb Ringer bicarbonate buffer before incubation. The islets of the pancreas were incubated in Kreb buffer under seven conditions of media treatment as follows: glucose medium that was used as a reference standard (R), total protein of seed (SPT), total protein of germinated soybean (GPT), TI protein of seed (SPTI), TI protein of germinated soybean (GPTI), KTI, and BBI. After two hours of incubation, the mixture was sonicated, and centrifuged The supernatant was stored at –20 oC for determination of insulin. The insulin analysis was conducted by ELISA method. The results show that aloksan injection increased the level of blood sugar and induced diabetic rats. The media treatment altered the soluble protein profile and the insulin secretion of the islets. The germinated SP has the ability to stimulate  insulin secretion of the pancreas of  diabetic as well as normal rats. The insulin secretion of the islets in the medium of crude TI of germinated soybean (GPTI) was highest. The potential order of the insulin secretion of normal rats islets were GPTI, KTI, BBI, GPT, SPTI, SPT, and R media. While the potential order of the insulin secretion of diabetic rat islets were GPTI, GPT, SPTI, BBI, KTI, SPT, and R media. The potential of germinated SP to stimulate insulin secretion was better than ungerminated SP.ABSTRAKTujuan dari penelitian ini adalah mempelajari potensi protein biji dan kecambah kedelai (protein total dan TI), Kunitz Trypsin Inhbitor (KTI), dan Bowman Birk Inhibitor (BBI)  dalam menstimulasi sekresi insulin pada pankreas tikus normal dan diabetes melalui pengujian biologis secara in vitro. Tikus Sprague Dawley (SD) jantan yang diguna- kan dalam pengujian biologis secara in vitro dibagi menjadi dua kelompok, yaitu tikus diabetes yang disiapkan dengan cara menginduksi melalui injeksi aloksan, dan tikus normal (tanpa diinjeksi aloksan). Selanjutnya tikus dieksekusi dengan bius ether dan dilakukan pencucian beberapa kali dengan larutan RPMI 1640 LITE. Pencucian terakhir meng- gunakan buffer Kreb Ringer Bicarbonat sebelum pancreas islet yang diperoleh diinkubasi. Pancreas islet diinkubasi dalam Kreb buffer yang dicampur dengan 7 perlakuan media inkubasi yang berbeda, yaitu medium glukosa sebagai control (R), protein total biji kedelai (SPT), protein total kecambah kedelai (GPT), protein TI biji kedelai (SPTI), pro- tein TI kecambah kedelai (GPTI), KTI dan BBI. Inkubasi dilakukan selama 2 jam, kemudian disonikasi, dan sentrifu- gasi. Supernatan yang diperoleh disimpan pada suhu -20 oC untuk disiapkan dalam  analisis insulin mengggunakan metode ELISA. Hasil penelitian menunjukkan bahwa injeksi aloksan pada tikus dapat menginduksi terjadinya diabetes karena dapat meningkatkan gula darah tikus. Perlakuan media inkubasi berpengaruh terhadap kadar protein terlarut media dan kemampuannya menstimulasi islet untuk mensekresikan insulin. Protein kecambah kedelai memiliki kemam- puan menstimulasi sekresi insulin baik pada pancreas tikus normal maupun diabetes. Sekresi insulin dari islet dalam medium GPTI paling tinggi dibandingkan perlauan yang lain. Urutan potensi sekresi insulin dari pancreas tikus nor- mal, yaitu berturut-turut dari yang tertinggi GPTI, KTI, BBI, GPT, SPTI, SPT, dan R. Sedangkan pada pancreas tikus diabetes, yaitu dari yang tertinggi GPTI, GPT, SPTI, BBI, KTI, SPT, and R media. Secara umum disimpulkan bahwa potensi protein kecambah kedelai dalam menstimulasi sekresi insulin lebih baik dibandingkan protein biji kedelai.