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All Journal EKSAKTA: Journal of Sciences and Data Analysis The Indonesian Biomedical Journal Molecular and Cellular Biomedical Sciences (MCBS) Jurnal Respirologi Indonesia Majalah Patologi Indonesia
Fadhillah, Muhamad Rizqy
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Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Dentistry Earth & Planetary Sciences Health Professions Immunology & microbiology Materials Science & Nanotechnology 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
The Role of Centella asiatica and Its Main Bioactive Compound, Asiatic Acid in Cardiac Remodeling: A Systematic Review of Animal Studies Fadhillah, Muhamad Rizqy; Arozal, Wawaimuli; Wibowo, Heri; Bustami, Arleni; Primadhani, Suci Widya; Gusti Khatimah, Nurul; Putri, Rizky Clarinta; Riski Amanda, Clara; Azizah, Nur
Molecular and Cellular Biomedical Sciences Vol 9, No 1 (2025)
Publisher : Cell and BioPharmaceutical Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21705/mcbs.v9i1.482

Abstract

Cardiac remodeling is a phenotype of heart failure characterized by a molecular, cellular, and interstitial change in the heart, which manifests in the change of size and function of the heart after specific insults with multiple mechanisms. Centella asiatica (CA) and its main bioactive triterpenoid, asiatic acid (AA), pose antioxidant and anti-inflammatory effects. Still, no adequate clinical trials support the potency of CA and AA as anti-cardiac remodeling. Hence, this systematic review aims to provide an in-depth analysis of CA extract and AA in animal studies' prevention or therapy of cardiac remodeling. The search strategies were conducted based on preferred reporting Items for systematic reviews and meta-analysis (PRISMA) protocol through Pubmed, EMBASE, Scopus, and Web of Science using keywords as follows: “Centella asiatica” OR “Asiatic Acid” AND “Cardiac Remodeling” OR “Cardiac Hypertrophy” OR “Cardiac Fibrosis” along with their synonym. The data collected included hemodynamic parameters based on echocardiography, biomolecular tests such as quantitative polymerase chain reaction (qPCR), Western blotting, or biochemistry procedures. The paper quality was assessed using Systematic Review Centre for Laboratory Animal Experimentation (SYRCLE) risk of bias (RoB). The previous selected study has shown that CA and AA might prevent and cure cardiac remodeling by inhibiting various pathways and protein expressions through AMPKα, NOX2/4, PI3K/Akt/mTOR, p70S6K, YAP/TAZ, and IL-1β, IL-6, and IL-18 cytokines. CA and AA, thus, exhibit cardioprotective effects in the animal model, which need to be confirmed in the clinical trials on humans. Keywords: cardiac remodeling, cardiac hypertrophy, cardiac fibrosis, Centella asiatica, asiatic acid
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
Optimasi Prosedur Kultur H9C2 Kardiomioblas: Sebuah Pembelajaran untuk Membangun Model Studi In-vitro Kardiovaskular Fadhillah, Muhamad Rizqy; Arozal, Wawaimuli; Wibowo, Heri
EKSAKTA: Journal of Sciences and Data Analysis VOLUME 5, ISSUE 1, April 2024
Publisher : Fakultas Matematika dan Ilmu Pengetahuan Alam

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20885/EKSAKTA.vol5.iss1.art8

Abstract

Cardiovascular disease become one of the leading factors of death in the world. Thus, research is urgently needed to discover newer drugs or therapeutical agents and biological plausibility. The H9C2 cardiomyoblast originated from embryonic BDIX rat ventricular cells and was previously used in numerous in vitro studies because of its similar nature to cardiomyocytes. However, to our knowledge, there are still limited studies on the basic procedure for culturing H9C2 cardiomyoblast and arranging the best strategy to perform a suitable timeline. Here, we shared our experience in culturing the H9C2 cardiomyoblast, including harvesting and subculturing the cells. We also demonstrated the change of cell confluency, depending on the seeding number, serum concentration, and culture flask through days 1, 3, and 6, to determine their doubling-time population. H9C2 cardiomyoblasts’ doubling time is around 48-54 days with Mean±SD 2.38±0.41. However, seeding density, different culture flasks, and serum concentration have become independent factors in determining specific measures to harvest the cells for further experiments.
Krebs von den Lungen-6 (KL-6) and Surfactant-D as Biomarkers of Interstitial Lung Disease: A Literature Review Fachrucha, Fanny; Fadhillah, Muhamad Rizqy; Aflah, Syazatul Syakirin Sirol
Jurnal Respirologi Indonesia Vol 44 No 1 (2024)
Publisher : Perhimpunan Dokter Paru Indonesia (PDPI)/The Indonesian Society of Respirology (ISR)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.36497/jri.v43i4.439

Abstract

Interstitial Lung Disease (ILD) is a group of lung diseases characterized by various patterns of lung tissue damage, including inflammation and fibrosis of the lung interstitium, both with known or unknown causes (idiopathic). In establishing the diagnosis of ILD, a comprehensive approach including history-taking, physical examination, and supporting examinations, is needed and managed in a multidisciplinary manner. Biomarkers are diagnostic tools known to be accessible, inexpensive, reproducible, and non-invasive for helping diagnose ILD patients. Growing evidence has supported the idea that many biomarker molecules can detect lung injury in ILD, including Krebs von de lungen-6 (KL-6) and Surfactant D (SP-D). KL-6 and SP-D could be utilized in the detection, disease monitoring, prognostication, and therapeutic responses of ILD patients. This review aimed to discuss several potential Kl-6 and SP-D biomarkers against ILD and discusses their clinical utility.
Co-Authors Aflah, Syazatul Syakirin Sirol Agian Jeffilano Barinda Antarianto, Radiana Dhewayani Arleni Bustami Dewi Sukmawati Fachrucha, Fanny Gusti Khatimah, Nurul Heri Wibowo Khatimah, Nurul Gusti Kusmardi Kusmardi Novi Silvia Hardiany Nur Azizah Primadhani, Suci Widya Putri, Rizky Clarinta Riski Amanda, Clara Shimizu, Ippei Triana, Novi Wawaimuli Arozal