Uji In-Vitro Translokasi GLUT4 pada Sel Otot Hiperinsulinemia terhadap Ekspresi Protein Akt dengan Blocker AS160

Izzah, Shabrina Ziyadatul (2026) Uji In-Vitro Translokasi GLUT4 pada Sel Otot Hiperinsulinemia terhadap Ekspresi Protein Akt dengan Blocker AS160. Other thesis, Institut Teknologi Sepuluh Nopember.

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Abstract

Diabetes Melitus Tipe 2 (DMT2) merupakan penyakit metabolik kronis yang ditandai oleh resistensi insulin. Salah satu jalur molekuler utama yang mengatur pengambilan glukosa adalah jalur pensinyalan insulin yang mengaktivasi transpor GLUT4 (Glucose Transporter Type 4). Pada kondisi hiperinsulinemia, insulin akan memicu aktivasi kaskade sinyal melalui protein INS (Insulin), INSR (Insulin Receptor), IRS-1 (Insulin Receptor Substrate), PI3K (Phosphoinositide 3-Kinase), PIP3 (Phosphatidylinositol (3,4,5)-trisphosphate), PDK1 (3-Phosphoinositide-dependent Protein Kinase-1) dan Akt (Protein Kinase B), tetapi dengan kondisi negative feedback yang menyebabkan penurunan sensitivitas insulin. Hal utama yang diuji pada penelitian ini adalah konsentrasi Akt dan GLUT4 yang terekspresi pada jalur pensinyalan insulin ketika dihambat maupun tidak dihambat oleh blocker AS160. Penelitian ini menggunakan sel otot spesifik myoblast human skeletal muscle (T48_CTF) yang diberi perlakuan hiperinsulinemia. Kemudian sel otot hiperinsulinemia dibagi menjadi 2 perlakuan; diberi blocker AS160 (P1) dan tidak diberi blocker AS160 (P2), dengan rentang waktu 10 menit hingga 50 menit. Selanjutnya, sel dikultur dan ekspresi protein total dihitung menggunakan ELISA reader. Hasil tersebut kemudian dianalisis menggunakan metode deskriptif. Hasil morfologi sel menunjukkan kondisi P2 memiliki kerusakan lebih parah dari P1. Konsentrasi protein Akt dan GLUT4 yang terekspresi pada P1 dan P2 memiliki persamaan pola kenaikan dan penurunan di menit awal yang selanjutnya mengalami perubahan respons insulin signaling yang tidak stabil. Akt pada P1 mengalami kenaikan konsentrasi, dengan kenaikan tertinggi dari 3,308 ± 0,176 mµ/L di menit 10 menjadi 14,529 ± 2,274 mµ/L di menit 20, lalu terjadi kenaikan sedikit di menit 30 dan 40, yaitu 15,488 ± 1,170 mµ/L dan 15,859 ± 0,663 mµ/L, dan turun kembali di menit 50 menjadi 14,466 ± 0,027 mµ/L. Akt pada P2 mengalami penurunan konsentrasi, berawal dari 19,241 ± 1,392 mµ/L di menit 10 menjadi 15,723 ± 7,322 mµ/L di menit 20, dan semakin turun pada menit 30 yaitu 9,279 ± 0,438 mµ/L, lalu meningkat di menit 40 yaitu 11,325 ± 1,899 mµ/L dan turun kembali pada menit 50 di angka 11,203 ± 0,981 mµ/L. GLUT4 pada P1 mengalami penurunan konsentrasi, namun diawali dengan kenaikan dari 1,562 ± 0,072 mµ/L di menit 10 menjadi 6,087 ± 0,844 mµ/L di menit 20, lalu meningkat sedikit menjadi 6,539 ± 0,478 mµ/L pada menit 30, selanjutnya menurun drastis menjadi 2,474 ± 0,582 mµ/L di menit 40 dan berakhir pada angka 1,302 ± 0,011 mµ/L di menit 50. GLUT4 pada P2 mengalami penurunan konsentrasi, dari 8,072 ± 0,569 mµ/L di menit 10 menjadi 5,565 ± 0,226 mµ/L di menit 20, lalu menjadi 4,002 ± 0,179 mµ/L di menit 30 hingga 1,555 ± 0,014 mµ/L di menit 40 dan selanjutnya mengalami peningkatan sedikit di menit 50 yaitu 2,378 ± 0,400 mµ/L. Pendekatan ini ditujukan untuk memperjelas peran relatif AS160 dalam modulasi efek Akt pada translokasi GLUT4 untuk merancang intervensi eksperimen yang lebih terarah pada studi resistensi insulin.
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Type 2 Diabetes Mellitus (T2DM) is a chronic metabolic disease characterized by insulin resistance. One of the main molecular pathways regulating glucose uptake is the insulin signaling pathway that activates GLUT4 (Glucose Transporter Type 4) transport. Under hyperinsulinemic conditions, insulin will trigger the activation of a signaling cascade through the proteins INS (Insulin), INSR (Insulin Receptor), IRS-1 (Insulin Receptor Substrate), PI3K (Phosphoinositide 3-Kinase), PIP3 (Phosphatidylinositol (3,4,5)-trisphosphate), PDK1 (3-Phosphoinositide-dependent Protein Kinase-1) and Akt (Protein Kinase B), but with a negative feedback condition that causes decreased insulin sensitivity. The main thing tested in this study was the concentration of Akt and GLUT4 expressed in the insulin signaling pathway when inhibited or not inhibited by the blocker AS160. This study used specific human skeletal muscle myoblast cells (T48_CTF) treated with hyperinsulinemia. Then, the hyperinsulinemic muscle cells were divided into two treatments: those treated with AS160 blocker (P1) and those not treated with AS160 blocker (P2), with a time span of 10 to 50 minutes. Next, the cells were cultured and total protein expression was calculated using an ELISA reader. The results were then analyzed using a descriptive method. The results of cell morphology showed that the P2 condition had more severe damage than P1. The concentrations of Akt and GLUT4 proteins expressed in P1 and P2 had similar patterns of increase and decrease in the initial minutes, which then experienced changes in the unstable insulin signaling response. Akt in P1 experienced an increase in concentration, with the highest increase from 3.308 ± 0.176 mµ/L at minute 10 to 14.529 ± 2.274 mµ/L at minute 20, then there was a slight increase at minute 30 and 40, namely 15.488 ± 1.170 mµ/L and 15.859 ± 0.663 mµ/L, and decreased again at minute 50 to 14.466 ± 0.027 mµ/L. Akt at P2 experienced a decrease in concentration, starting from 19,241 ± 1,392 mµ/L at minute 10 to 15,723 ± 7,322 mµ/L at minute 20, and decreased further at minute 30, namely 9,279 ± 0,438 mµ/L, then increased at minute 40, namely 11,325 ± 1,899 mµ/L and decreased again at minute 50 at 11,203 ± 0,981 mµ/L. GLUT4 in P1 experienced a decrease in concentration, but it started with an increase from 1.562 ± 0.072 mµ/L at minute 10 to 6.087 ± 0.844 mµ/L at minute 20, then increased slightly to 6.539 ± 0.478 mµ/L at minute 30, then decreased drastically to 2.474 ± 0.582 mµ/L at minute 40 and ended at 1.302 ± 0.011 mµ/L at minute 50. GLUT4 in P2 experienced a decrease in concentration, from 8.072 ± 0.569 mµ/L at minute 10 to 5.565 ± 0.226 mµ/L at minute 20, then to 4.002 ± 0.179 mµ/L at minute 50. mµ/L at minute 30 to 1.555 ± 0.014 mµ/L at minute 40 and then increased slightly at minute 50 to 2.378 ± 0.400 mµ/L. This approach aimed to clarify the relative role of AS160 in modulating the effect of Akt on GLUT4 translocation in order to design more targeted experimental interventions in the study of insulin resistance.

Item Type:	Thesis (Other)
Uncontrolled Keywords:	Akt, DMT2, GLUT4, Resistensi Insulin, Translokasi, Akt, GLUT4, Insulin Resistance, TDM2, Translocation
Subjects:	Q Science > QH Biology
Divisions:	Faculty of Science and Data Analytics (SCIENTICS) > Biology > 46201-(S1) Undergraduate Thesis
Depositing User:	Shabrina Ziyadatul izzah
Date Deposited:	01 Aug 2026 03:52
Last Modified:	01 Aug 2026 03:52
URI:	http://repository.its.ac.id/id/eprint/139658

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