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Optimized RT-qPCR Detection of Hepatic Lipopolysaccharide-Binding Protein in Diet-Induced Obese Mice Tjahjono, Yudy; Wedharga, I Gede Putu Adhi; Novita, Bernadette Dian; Tahalele, Paul; Wijaya, Hendy; Soediono, Endang Isbandiati; Hendrata, Adi Pramono; Dewi, Sianty; Wijaya, Sumi; Ervina, Martha; Kuncorojakti, Suryo
Jurnal Medik Veteriner Vol. 8 No. 2 (2025): October
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/jmv.vol8.iss2.2025.308-317

Abstract

High-throughput RT-qPCR results on hepatic lipopolysaccharide-binding protein (LBP) expression in obese subjects are essential, as they reveal the endotoxin’s role in the development of obesity and non-communicable disease (NCD). This study aimed to optimize RT-qPCR detection of LBP in diet-induced obese mice. This study primarily focused on addressing high variability through reference gene normalization. A total of six male C57BL/6 mice aged 6 weeks were randomly allocated into two dietary treatments (n = 3), consisting of mice fed with the standard chow diet (SCD group) and mice fed with the high-fat and high sucrose diet (HFHS group) ad libitum for 8 weeks. Relative quantification strategies involving the standard 2-ΔΔCt method (calibrator as mean) and the modified 2-ΔΔCt method (calibrator as individual sample-matched biological replicates) were compared in terms of their variability. Obesity was successfully induced in the HFHS treatment group, as indicated by significantly higher body weight, calorie intake, and LBP relative expressions compared to the SCD group. In addition, a sample-specific calibrator approach using the modified 2-ΔΔCt method resulted in lower variability in relative gene expression levels. A modified 2-ΔΔCt method, which utilizes a sample-specific calibrator to counteract sample-specific variability, was successfully employed to address high variability in RT-qPCR results.
HYPOGLYCEMIC, ANTI-INFLAMMATORY EFFECT OF PORANG (AMORPHOPHALLUS ONCHOPYLLUS) ON ALLOXAN-INDUCED DIABETIC RATS Young, Andre; Novita, Bernadette Dian; Ricardo, Evan; Dewi, I.G.A.P.D; Amadeo, Jovan; Dewi, Sianty; Tahalele, Paul
JURNAL WIDYA MEDIKA Vol. 9 No. 1 (2023): April
Publisher : FAKULTAS KEDOKTERAN UNIVERSITAS KATOLIK WIDYA MANDALA SURABAYA

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33508/jwm.v9i1.4173

Abstract

Introduction: Iles-iles / Porang is a tuber-producing plant that is commonly found in Indonesia. One of the most sought after content from Porang is Glucomannan. One of Porang’s health benefit is related to the effect of lowering blood glucose levels because it can prevent glucose absorption. These benefit is interesting to study because there has been no research linking the use of Porang to reduce inflammatory process in hyperglycmic conditions. Purpose: This study aims to analyze the anti-inflammatory, anti-oxidant and hypoglycemis effects of Glucomannan from Porang (Amorphophallus onchophyllus) extract in experimental animals. Method: The research design was a true experimental post-test only control group with random sampling to determine 5 white rats into the normal group, positive control group, negative control group and treatment 1, 2 and 3. The positive control group received Acarbose therapy thile the negative control group received carboxy-methyl-cellulose (CMC) therapy. This study used Porang extract (Amorphophallus onchophyllus) with concentrations of 200, 400 and 800 mg/Kg in hyperglycemic white rats that had been induced by Aloxan. The study was conducte for 50 days and then blood and serum samples were taken to assess the effect of hypoglycemia, anti-inflammatory and anti-oxidant using blood glucose, Malondialdehyde and C-reactiveprotein (CRP) measurement. Result and Discussion: The results showed no significant difference between the groups of rats receiving Porang extract and the positive and negative control groups. However, the MDA levels after 50 days of intervention between the negative control group and the therapy with doses of 200, 400 and 800 mg/Kg showed significant differences. Similarly found for blood glucose levels after intervention between negative control group and the 200 and 400 mg/Kg therapy group. This results may be caused by the type of Porang used, the form of the Porang and the concentration level of the Porang extract. Conclusion: Porang with the type of Amorphophallus onchophyllus can’t be used directly, but requires further processing to obtain the active substance Glucomannan.
Modified High-Fat High-Sucrose Diet Promotes Obesity and Alters Colonic Cytokines Novita, Bernadette Dian; Wedharga, I Gede Putu Adhi; Tjahjono, Yudy; Wijaya, Hendy; Theodora, Imelda; Ervina, Martha; Wilianto, Yufita Ratnasari; Dewi, Sianty; Parengkuan, Irene Lingkan; Herjunianto, Herjunianto; Ghasani, Sabrina Maria; Limantoro, Michael Christian; Jaya, Ferdinand Wiliam
The Indonesian Biomedical Journal Vol 17, No 5 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i5.3807

Abstract

BACKGROUND: Western dietary patterns are often associated with increasing non-communicable diseases, including inflammatory bowel disease (IBD). In experimental models, a high-fat high-sucrose (HFHS) diet is used to mimic metabolic and inflammatory effects of such diets, however the data regarding colonic inflammation in Indonesia remain limited. Therefore, this study was conducted evaluated the impact of HFHS feeding on colonic interleukin (IL)-6, IL-10 expression, and the IL-6/IL-10 ratio.METHODS: Six weeks old male C57BL/6 mice were randomly assigned to a normal fat diet (NFD) or HFHS diet group and fed ad libitum for 8 weeks. Colonic tissues were collected, and IL-6 and IL-10 expression was analyzed by immunohistochemistry.RESULTS: HFHS-fed mice showed significant increases in body weight (increased by 22.44%, p=0.0047) and caloric intake (increased by 125.17%, p=0.0000), confirming obesity induction. Colitis was also evident, with higher histological colitis scores (p=0.0072). However, colonic IL-6 (increased by 9.12%, p=0.1236), IL-10 (increased by 1.49%, p=0.8013), and the IL-6/IL-10 ratio (increased by 7.38%, p=0.4000) showed no significant differences compared to NFD.CONCLUSION: In C57BL/6 mice, an 8-week modified HFHS diet induced obesity, increased caloric intake, and mucosal injury, but did not significantly alter colonic IL-6, IL-10, or their ratio. This suggests preserved mucosal immune homeostasis consistent with an early compensatory phase rather than overt cytokine-driven inflammation. Longer or more intensive exposure may disrupt this balance, highlighting the need for further studies to define the temporal threshold and clarify immune microbiome interactions in colitis progression.KEYWORDS: high-fat high-sucrose diet, colon inflammation, IL-6, IL-10, obesity mice
Tumor Differentiation is Correlated with Estrogen Receptor Beta (ERβ) Expression but Not with Interleukin-6 (IL-6) Expression in Colorectal Carcinoma Theodora, Imelda; Sudiana, I Ketut; Budipramana, Vicky Sumarki; Erwin, Ferdinand; Dewi, Sianty; Novita, Bernadette Dian
The Indonesian Biomedical Journal Vol 16, No 5 (2024)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v16i5.3324

Abstract

BACKGROUND: Colorectal carcinoma (CRC), the third most common malignant disease worldwide, is associated with estrogen receptor β (ERβ) and interleukin-6 (IL-6). ERβ is known to down-regulate IL-6 in prostate cancer, lung carcinoma, and CRC cell lines; however, its effect on human with CRC remains unclear. Therefore, this study was conducted to investigate the association between ERβ and IL-6 expressions with the clinicopathological features of CRC.METHODS: This was an analytic observational study using 40 paraffin blocks of CRC patients. ERβ and IL-6 expression was measured by immunohistochemistry (IHC) staining. The percentage of immunoreactive tumor cell per 1000 cells was manually recorded and tumor differentiation as well as tumor infiltration were determined. Tumor differentiation was graded according to the World Health Organization (WHO) 2010 criteria, while tumor infiltration was defined based on the American Joint Committee on Cancer (AJCC) 8th edition.RESULTS: Fifty percent of samples were well-differentiated CRC, and 57.5% samples were T3 infiltration tumors. IHC staining showed 35.5% of samples were positive for ERβ expression, while 70.86% were positive for IL-6 expression. There were negative correlation of ERβ expression with tumor differentiation (p=0.018; r=-0.371), but no correlation with tumor infiltration (p=0.836) were found. There was no correlation between ERβ expression with IL-6 expression (p=0.154).CONCLUSION: There is statistically significant correlation between tumor differentiation and ERβ expression, wherein improved tumor differentiation is linked to higher levels of positive ERβ expression. However, there is no discernible relationship between IL-6 and tumor differentiation. These findings suggest that while IL-6 was involved in the growth of the tumor, ERβ expression might have an impact on tumor differentiation.KEYWORDS: colorectal carcinoma, estrogen receptor beta, interleukin-6, cell differentiation