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The Effect of Increased Glucose Induction on GSH Levels in Insulin Gaussia Luciferase (iGL) Cells Derived from Rat Pancreatic Beta Cells Ardiansah, Fery; Stujanna, Endin Nokik; Sanjaya, Arief Indra; Listiyaningsih, Erlin; Ningsih, Sri Suciati; Ujianti, Irena; Lestari, Dwi Retna
Journal of Biomedicine and Translational Research Vol 9, No 3 (2023): December 2023
Publisher : Faculty of Medicine, Universitas Diponegoro

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/jbtr.v9i3.19644

Abstract

Background: Prolonged hyperglycaemia can make the pancreatic beta cells work harder and cause fatigue. When this happens, it can trigger oxidative stress reactions, which can produce free radical compounds that can damage pancreatic beta cells. The body compensates by activating protective mechanisms such as the production of antioxidant compounds to reduce the levels of free radicals in the cells. One such compound is glutathione (GSH). Insulin Gaussia Luciferase (iGL) cells are a cell line derived from rat pancreatic β (beta) cells. These cells can be used as a model of oxidative stress in hyperglycaemia to measure GSH levels and there are no studies using iGL cells to measure GSH levels. Therefore, in this study, the iGL cells are used as the object of research.Objective: To investigate the effect of GSH levels on glucose toxicity condition through in vitro experiments on iGL cells.Methods: The study used 5 different glucose concentrations of 11, 16.5, 22, 33, and 44 mM with the addition of iGL cell growth medium exposed for 7 days. We measured the amount of intracellular GSH using a colourimetric method at a wavelength of 405 nm. The analysis used in this study was a one-way ANOVA test. Differences between groups were tested using SPSS.Results: The results of this study showed that there was an increasing trend in total GSH levels on the third and seventh day.Conclusion: there was a daily decrease in GSH/cell in the iGL sample cells exposed to different concentrations of glucose for 7 days. This can be due to an increase in the oxidative stress reactions in the cells, which can lead to a decrease in the levels of antioxidants.
Computational study of active compounds of Citrullus lanatus Linn peel extract as potential antidiabetics Sari, Shinta Dewi Permata; Budiman, Muhamad Arif; Ningsih, Sri Suciati
Indonesian Journal of Biomedicine and Clinical Sciences Vol 57 No 2 (2025)
Publisher : Published by Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/inajbcs.v57i2.14518

Abstract

Diabetes mellitus (DM) is one of the metabolic diseases that have emerged as a global health problem. Type 2 diabetes mellitus (T2DM), which affect 90-95% of DM, is caused by reduced insulin sensitivity and insulin resistance in peripheral tissues. Oral antidiabetics have resulted side effects, prompting an investigation for a natural-based antidiabetic agent as an alternative treatment. Using network pharmacology, we investigated the mechanism of phytochemical substances of Citrulus lanatus Linn. peel extract and their interactions with target proteins in the DM pathogenesis pathway. Cytoscape 3.6.1 software has created a network of extract compound-protein targets. Investigation of protein interaction, target gene function enrichment, and signal pathway performed via DAVID, STRING database, and the KEGG pathway database. The computational study identified 90 target proteins associated with T2DM based on protein-protein interactions. In addition, Cytoscape analysis and DAVID enrichment revealed the network of extract compound's target and generated proteins such as INS, TNF-α, IL-6, and AKT2. The KEGG pathway analysis presented the crucial role of insulin resistance and AGE-RAGE signaling pathways. This pathway correlated with lower glucose activity in obesity and hyperglycemia. It indicates that the active constituents of C. lanatus Linn peel extract can lower blood sugar levels by interacting with selected proteins. This study's findings will be carried out in further research of in vitro trials.
Immunocytochemistry Evaluation of Fibroblast Response to Garlic (Allium sativum) Extract Targeting MMP9: In Vitro and In Silico Approaches Ningsih, Sri Suciati; Zuraidah, Endah; Handayani, Supri Irianti; Ardiansyah, Ahmad Ali Irfan; Wahyuni, Ningrum
Indonesian Journal of Medical Chemistry and Bioinformatics
Publisher : UI Scholars Hub

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Background: Fibrosis involves excessive extracellular matrix (ECM) deposition, with matrix metalloproteinase-9 (MMP-9) playing a critical role. Natural compounds targeting MMP-9 may offer therapeutic potential. Objective: This study evaluated the antifibrotic effect of Allium sativum extract on fibroblast cells through in vitro and in silico approaches targeting MMP-9. Methods: Human dermal fibroblasts were treated with A. sativum extract (25, 50, and 100 µg/mL) for 24 and 48 hours. Cell viability was assessed using MTT assay. MMP-9 expression was analyzed via quantitative real-time PCR (qRT-PCR) and immunocytochemistry (ICC). Major garlic compounds were docked to MMP-9 using AutoDock Vina. Results: The 50 µg/mL concentration maintained >85% cell viability and significantly reduced MMP-9 gene and protein expression. S-allyl cysteine exhibited the strongest binding affinity (−6.8 kcal/mol) and formed hydrogen bonds with key residues (Glu402, Ala189, His405) of MMP-9. Conclusion: Allium sativum extract demonstrates antifibrotic activity by downregulating MMP-9 expression and inhibiting its function, supporting its potential as a natural MMP-9 inhibitor for fibrosis-related disorders.
Specific Primer Design to Detect Connexin-36 Gene in Rattus Norvegicus Brain Ningsih, Sri Suciati; Septiasari, Alfina; Stujanna, Endin Nokik; Avissa, Rizkyana; Ujianti, Irena; Permatasari, Shinta Dewi; Sukarya, Wawang S.
THE JOURNAL OF MUHAMMADIYAH MEDICAL LABORATORY TECHNOLOGIST Vol 7 No 1 (2024): The Journal of Muhammadiyah Medical Laboratory Technologist
Publisher : Universitas Muhammadiyah Surabaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.30651/jmlt.v7i1.17985

Abstract

Connexin-36 (Cx36) merupakan salah satu protein kanal antar sel. Protein ini paling banyak ditemukan pada otak. Gen Cx36 mengekspresikan protein penghubung Cx36 yang membentuk sinaps listrik. Penelitian ini bertujuan untuk memperoleh desain primer terbaik untuk menganalisis ekspresi Cx36 pada otak tikus dengan metode Quantitative Real Time PCR (qRT-PCR). Jenis penelitian ini adalah penelitian true experimental menggunakan mRNA dari 4 otak tikus. Analisis in silico menggunakan tools Primer 3 dan OlygoAnalizerTM by IDTdengan menggunakan database gen pada NCBI. Tiga primer dengan kriteria yang paling optimum kemudian dievaluasi dengan metode qRT-PCR. Berdasarkan hasil penelitian ketiganya mampu mengamplifikasi gen Cx36 dengan menggunakan qRT-PCR. Hasil primer terbaik menunjukkan melt curve tunggal pada primer no.5 gen Cx36 pada kisaran Tm 80-81oC. Hasil perbandingan uji in silico primer Cx36 yang sudah pernah dipublikasi sebelumnya menunjukkan bahwa primer no. 5 juga memiliki nilai optimal pada setiap kriteria. Maka hasil desain primer yang memiliki spesifisitas dan efektifitas yang optimum untuk mengamplifikasi gen Cx36 pada otak tikus Rattus norvegicus ialah primer no.5 (F: 5’-ATTTCCCGCTTCTACATCATCCAAG-3’ dan R: 5’-CACAGCAAACATGAACACCAGAAAG-3’).