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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