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All Journal The Indonesian Biomedical Journal Molecular and Cellular Biomedical Sciences (MCBS) Majalah Patologi Indonesia Majalah Biomorfologi (Biomorphology Journal)
Khatimah, Nurul Gusti
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Biochemistry, Genetics & Molecular Biology Dentistry Health Professions Immunology & microbiology Medicine & Pharmacology Neuroscience Public Health

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Hypertrophy Determination of H9c2 Cardiomyoblast Cell Line Using Wright-Giemsa Staining: An Experience in Developing an Acceptable and Easy-to-handle In-vitro Protocol Fadhillah, Muhamad Rizqy; Arozal, Wawaimuli; Wibowo, Heri; Bustami, Arleni; Sukmawati, Dewi; Kusmardi, Kusmardi; Triana, Novi; Khatimah, Nurul Gusti
Majalah Patologi Indonesia Vol. 33 No. 2 (2024): MPI
Publisher : Perhimpunan Dokter Spesialis Patologi Anatomik Indonesia (PDSPA)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.55816/mpi.v33i2.682

Abstract

Introduction Cell-size area (CSA) becomes the standard parameter routinely tested in vitro for cardiac hypertrophy studies. Thus, staining is an essential tool for this purpose. As reported in a previous study, immunofluorescence staining is an established method for CSA. However, because it is expensive and requires a specialized microscope, e.g., immunofluorescence or confocal microscope, it is not applicable in a laboratory with limited equipment. Wright-Giemsa staining is a standard procedure in hematology laboratories and is inexpensive and convenient. Here, we shared our experience developing a CSA determination protocol using Wright-Giemsa in H9c2 cardiomyoblast. Methods The viability tests were performed on H9c2 to determine the effective dosage of angiotensin II and Irbesartan (standard drug). The H9c2 were divided into three groups: the control group (without either angiotensin II or irbesartan), the negative control (with angiotensin II), and the positive control (with angiotensin II and Irbesartan), triplicate for each group. The cells then are acclimatized overnight, serum-starved for one day, and incubated with angiotensin and irbesartan for 48 hours. Lastly, Wright-Giemsa was observed using a light microscope in three fields. The CSA was determined by three independent observers blindly, statistically different if the p<0.05 using ANOVA ways or Kruskal-Wallis. Results After the H9c2 induced by angiotensin-II 1 μM and Irbesartan 1μM, we found the CSA significantly differed among each group (p<0,0001). The negative control has a higher median and interquartile range (IQR) CSA (10.78 (6.79) um2) compared to the control group (median (IQR) 7.27 (4.91) um2) and positive control (median (IQR) 7.849(5.31) um2). Conclusion It can be concluded that the Wright-Giemsa might help determine the CSA for in-vitro hypertrophic studies.
Potential Anti-Senescence Effect of Extract from Andrographis paniculata Herbal Plant and Its Bioactive Compounds: A Systematic Review Khatimah, Nurul Gusti; Arozal, Wawaimuli; Barinda, Agian Jeffilano; Antarianto, Radiana Dhewayani; Hardiany, Novi Silvia; Shimizu, Ippei; Fadhillah, Muhamad Rizqy
Molecular and Cellular Biomedical Sciences Vol 8, No 3 (2024)
Publisher : Cell and BioPharmaceutical Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21705/mcbs.v8i3.432

Abstract

The rapid aging of the global population is a major worldwide issue because of the close relationship between age and the development of several diseases. Aging or senescence is among the most widely studied topics at the moment. However, no pharmaceuticals have been developed that claim to possess anti-senescence properties. Andrographis paniculata, is a medicinal plant found widely throughout tropical and subtropical Asia. This review aims to identify the potential anti- senescence effect of A. paniculata extract and its bioactive compounds. By following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, five databases were used and in vivo and in vitro studies were included in this review. A. paniculata extracts and their bioactive compounds exert anti-senescence properties through their anti-inflammatory and antioxidant properties. This herb and its compounds enhanced memory, cognitive function and behaviour in Alzheimer's disease. The extract also promoted cell cycle progression and proliferation in the skin. In addition, andrographolide exhibited anti-senescence effects in endothelial cells through the activation of PI3K/Akt/Nrf and PI3K/Akt/AP-1 pathways. A. paniculata along with its bioactive compounds including andrographolide and 14-deoxyandrographolide, may have the potential to be used as anti-senescence through anti-inflammatory and antioxidant properties. However, the specific markers to evaluate the senescence are necessary to be conducted. Any clinical trials should be done to establish these findings. Since in clinical settings this potential herbal may be used for long-life time, the safety profile and toxicity of A. paniculata should be considered. Keywords: herbal plants, Andrographis paniculata, andrographolide, bioactive compounds, senescence
Investigating the Prospect of C-Type Lectin-like Receptors-2 for Predicting Prognosis of Ischemic Stroke: Rapid Review of Molecular Mechanisms to Clinical Study Farandy, Rayhan; Rakasiwi, Muhammad Ilham Dhiya; Amaanullah, Muhammad Zaki Bariz; Khatimah, Nurul Gusti
Majalah Biomorfologi Vol. 35 No. 2 (2025): Majalah Biomorfologi
Publisher : Universitas Airlangga, Surabaya, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/mbiom.v35i2.2025.157-165

Abstract

Highlights The potential of CLEC-2 as a prognostic biomarker for ischemic stroke is supported by in vivo and clinical studies demonstrating its role in thrombo-inflammatory mechanisms and clinical outcome prediction. Analysis of clinical studies indicates that elevated plasma CLEC-2 levels are associated with an increased risk of recurrent vascular events and mortality in acute ischemic stroke, suggesting its promise as a noninvasive tool for stroke prognosis assessment.   ABSTRACT Background: Every year, over 13.7 million individuals experience strokes, resulting in approximately 5.8 million deaths. C-type lectin-like receptor 2 (CLEC-2) plays a significant role in platelet activation, which is elevated in ischemic stroke and is associated with disease progression and prognosis. Objective: To review the literature on the potential of CLEC-2 as a biomarker for assessing the prognosis and progression of ischemic stroke. Material and Method: This rapid review followed the Cochrane interim guidelines and adhered to PRISMA standards. A comprehensive search was conducted in PubMed, Cochrane Library, and Google Scholar to identify original research articles published in English over the past 10 years. Studies at various stages—including in vitro, in vivo, and clinical trials—were included if they evaluated the association between CLEC-2 and acute ischemic stroke. Risk of bias was assessed using the QUIPS tool for clinical studies and SYRCLE’s tool for animal studies. Study selection and data extraction were performed independently by three reviewers. Result: The search identified five relevant articles: two experimental studies and three clinical prognostic studies examining CLEC-2 in the context of ischemic stroke. CLEC-2, a receptor for podoplanin expressed in various tumors and lymphatic endothelial cells, induces a calcium surge independent of secondary platelet activation. In vivo studies have demonstrated increased levels of CLEC-2 and podoplanin, which are highly expressed on neurons and microglia in ischemic brain regions. The three clinical studies showed that plasma CLEC-2 levels have prognostic value in predicting recurrent vascular events and mortality in patients with acute ischemic stroke. Conclusion: Plasma CLEC-2 shows potential as a biomarker for evaluating the progression and prognosis of acute ischemic stroke.
Andrographis paniculata Ethanol Extract Alleviates High Glucose-induced Senescence of Human Umbilical Vein Endothelial Cells via the Regulation of mTOR and SIRT1 Pathways Khatimah, Nurul Gusti; Arozal, Wawaimuli; Barinda, Agian Jeffilano; Antarianto, Radiana Dhewayani; Hardiany, Novi Silvia; Shimizu, Ippei; Fadhillah, Muhamad Rizqy
The Indonesian Biomedical Journal Vol 16, No 4 (2024)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v16i4.3067

Abstract

BACKGROUND: Chronic exposure of high glucose (HG) in endothelial cell induces senescence which may contribute to the development and progression of age-related diseases including insulin resistance. Andrographis paniculata improves insulin resistance in recent in vitro and in vivo studies. Anti-inflammatory and antioxidant properties of A. paniculata may be the new therapeutic approach to inhibiting premature senescence. However, the senolytic effect of A. paniculata on endothelial cells has not been investigated comprehensively. This study was conducted to evaluate the effect of A. paniculata extract on HG-induced endothelial cell senescence and the underlying mechanisms.METHODS: Human umbilical vein endothelial cells (HUVECs) were treated with 33 mM HG and 7.5 μg/mL A. paniculata extract for 48 hours. The expressions of p16, p21, interleukin (IL)-6, IL-8, insulin receptor substrate (IRS)-1, mammalian target of rapamycin, and sirtuin 1 (SIRT1) were measured by performing real-time quantitative polymerase chain reaction (RT-qPCR). The senescence-associated-β-galactosidase (SA-β-gal) staining was performed to observe the positive-stained senescent cells, while the cell surface expression of IL-1α was examined with flow cytometry method.RESULTS: A. paniculata extract reversed senescence in HUVECs under HG conditions by reducing mRNA expressions of p16 and p21, the number of SA-β-gal-positive-stained cells, and the expression of IL-1α on cell surface, which decreased the activation of IL-6 and IL-8. In addition, A. paniculata extract decreased the mRNA expression of mTOR and increased the mRNA expressions of IRS-1 as well as SIRT1.CONCLUSION: A. paniculata extract ameliorated senescence and improved insulin sensitivity by regulating the mTOR, SIRT1, and IRS-1 mRNA expressions on HG-treated HUVECs.KEYWORDS: Andrographis paniculata, endothelial cell, senescence, high glucose, nutrient-sensing pathways
Co-Authors Agian Jeffilano Barinda Amaanullah, Muhammad Zaki Bariz Antarianto, Radiana Dhewayani Arleni Bustami Dewi Sukmawati Fadhillah, Muhamad Rizqy Farandy, Rayhan Heri Wibowo Kusmardi Kusmardi Novi Silvia Hardiany Rakasiwi, Muhammad Ilham Dhiya Shimizu, Ippei Triana, Novi Wawaimuli Arozal