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Hesperidin increase cytotoxic effect of doxorubicin in MCF-7 cells Hermawan, Adam; Meiyanto, Edy; Susidarti, Ratna Asmah
Indonesian Journal of Pharmacy Vol 21 No 1, 2010
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

Hesperidin,Â aÂ flavonoid,Â showsÂ strongÂ cytotoxicÂ effectÂ in Â severalÂ cancer cellÂ lines.Â TheÂ aimÂ ofÂ thisÂ researchÂ wasÂ toÂ investigateÂ cytotoxicÂ activitiesÂ of hesperidinÂ aloneÂ andÂ inÂ combinationÂ withÂ doxorubicin.Â CellÂ viabilityÂ assayÂ of hesperidin,Â doxorubicin,Â andÂ combinationÂ treatmentsÂ wereÂ carriedÂ outÂ byÂ using MTTÂ assay.Â ApoptosisÂ assayÂ wasÂ doneÂ usingÂ doubleÂ stainingÂ methodÂ using EthidiumÂ Bromide-AcridineÂ Orange.Â HesperidinÂ didÂ notÂ showÂ cytotoxicÂ effect but doxorubicin showed cytotoxic effect with IC50467 nM. Hesperidin (5, 50 and 100Â ÂµM)Â increasedÂ cytotoxicÂ effectÂ ofÂ doxorubicinÂ comparedÂ withÂ doxorubicin alone.Â TheÂ strongestÂ cytotoxicÂ activityÂ wasÂ showedÂ byÂ theÂ combinationÂ ofÂ 200 nMÂ doxorubicinÂ andÂ 100Â ÂµMÂ hesperidin.Â CombinationÂ treatmentÂ ofÂ doxorubicin 200Â nMÂ andÂ hesperidinÂ 100Â ÂµMÂ inducedÂ apoptosisÂ inÂ MCF-7 cells.Â HesperidinÂ is potentiallyÂ toÂ beÂ developedÂ asÂ co-chemotherapeuticÂ agentÂ forÂ breastÂ cancer, while molecular mechanism need to be explored.Key words: Hesperidin, doxorubicin, synergism, MCF-7, apoptosisÂ

Visible spectrophotometric determination of Cefadroxil using ethyl acetoacetate and formaldehyde reagents Susidarti, Ratna Asmah; Rianti, Andrih; Martono, Sudibyo
Indonesian Journal of Pharmacy Vol 19 No 1, 2008
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

Visible spectrophotometric method can be used to determine cefadroxil in the pharmaceutical dosage form. The reaction of cefadroxil with the condensation product of 2 mol ethyl acetoacetate and 1 mol formaldehyde in acidic condition (pH 3,5) at 45Â°C for 20 minutes yielded yellow product giving absorption at 367 nm. The concentration of Cefadroxil was calculated by using standard curve Y = 1.6063 X + 0.1634, r = 0.9932, p = 0,001 and Vxo = 9,7%. Determination of cefadroxil capsule â€Xâ€ (562.4 mg/capsule) showed that the concentration of cefadroxil observed is 490.5 mg/capsule (CV = 0.89%) and recovery value is 102.88% (CV = 0.67%). Key words: visible spectrophotometry, Cefadroxil, ethyl acetoacetate, formaldehyde

Antioxidant–free radical scavenging of flavonoid from The Leaves of Stelechocarpus burahol (Bl.) Hook f. & Th. Sunarni, Titik; Pramono, Suwidjiyo; Asmah, Ratna
Indonesian Journal of Pharmacy Vol 18 No 3, 2007
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

The activity testing of flavonoid compounds as antioxidant and as scavenger of free radical, isolated from the Stelechocarpus burahol leaves had been performed. The aqueous extract of the leaves was concentrated and then suspended in ethanol to produce ethanolic fraction. The fraction was chromatographed on several paper chromatography systems to produce isolate with chromatographic purity.The all isolated flavonoid was identified by paper chromatography system. Especially, the B4b isolate were identified further using spectrometer UV-Vis, FT-IR and ¹H-NMR. Their antioxidant activities were done by the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging method.The all isolated flavonoid showed activity as DPPH scavenger. Among these isolate, B4b exhibited a strong free radical scavenging with an EC50 value of 6.43 μg/mL. They were identified and B4b isolate was pressumed as flavon with hydroxyl group on C-3, C-7, C-3', C-4' and methyl on C-5.Key words : Stelechocarpus burahol, flavonoid, antioxidant, DPPH

DPPH Scavenging Activity, Reducing Power, and Metal Chelating Capacity of Compound 1-(2,5-dihydroxyphenyl)-3-pyridine-2-yl-propenone Octa Ujiantari, Navista Sri; Wibowo, Andy Eko; Susidarti, Ratna Asmah
Journal of Food and Pharmaceutical Sciences Vol 11, No 3 (2023): J.Food.Pharm.Sci
Publisher : Integrated Research and Testing Laboratory (LPPT) Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jfps.7317

Background: The excessive free radicals in the body could stimulate oxidative stress which has been implicated in several diseases such as cancer, inflammation, aging, and cardiovascular diseases. The presence of antioxidants can diminish the reactivity of free radicals. The antioxidant defense system in the body may be insufficient thus intake of dietary antioxidants is recommended. Natural flavonoid such as chalcone has been known to exert several biological activities, especially antioxidant activity. Compound 1-(2,5-dihydroxyphenyl)-3-pyridine-2-yl-propenone was successfully synthesized using microwave radiation and several studies reported its biological activities such as antiinflammation, anticancer, and antibacterial activities. Those activities are thought to be related to antioxidant mechanisms. Therefore, this study aimed to evaluate its antioxidant activity. Methods: The antioxidant activities were performed with three different methods: DPPH scavenging activity, ferric reducing power, and metal chelating capacity. Results: 1-(2,5-dihydroxyphenyl)-3-pyridine-2-yl-propenone showed DPPH scavenging activity, ferric reducing power, and metal chelating capacity with IC50S values of 4,471 ± 0,052 μg/mL; 156.56 ± 4.42μg/mL; and 6,273 ± 0,025 μg/mL, respectively. Conclusions: Our results found that 1-(2,5-dihydroxyphenyl)-3-pyridine-2-yl-propenone had quite potent antioxidant activity.

Comparative Analysis of Essential Oil Profiles From Emprit Ginger Rhizome (Zingiber officinale var. amarum) Grown in Different Locations and Antibacterial Activity Againts Staphylococcus aureus Styawan, Anita Agustina; Purwanto; Susidarti, Ratna Asmah; Windarsih, Anjar; Rohman, Abdul
Indonesian Journal of Tropical and Infectious Disease Vol. 12 No. 2 (2024)
Publisher : Institute of Topical Disease Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/ijtid.v12i2.50423

Indonesia, as a tropical country, boasts significant biodiversity, particularly regarding its flora and fauna. The country's medicinal plants are one inherited natural resource passed down through generations. The "emprit ginger” (Zingiber officinale var. amarum), a native Indonesian medicinal plant, is often employed in treating various diseases. Scientifically, the primary constituent of emprit ginger is the phenolic ketone homolog compound, gingerol, known for its antioxidant activity. In addition to its antioxidant potential, emprit ginger exhibits potential as an antibacterial agent. However, herbal substances used for therapeutic purposes often yield inconsistent effects due to the fluctuating chemical composition in the plants, typically a result of varying growing locations. These location differences can cause discrepancies in the content of active metabolites. Therefore, data on the chemical profile of medicinal plants is paramount for the standardization process. This study was conducted by obtaining essential oils from the rhizomes of emprit ginger, extracted via steam distillation obtained from various cultivation lands in Ponorogo, Magetan, Pacitan, Wonogiri, Karanganyar, Boyolali, Semarang, Magelang, Purworejo, Temanggung, Wonosobo, Banyumas, Bantul, and Kulonprogo. One of the efforts in standardizing natural materials involves the analysis of their metabolite profiles. The emprit ginger essential oil profile was obtained through Gas Chromatography-Mass Spectrometry (GCMS) testing. The emprit ginger essential oil profile was analyzed using a multivariate calibration of Principal Component Analysis utilizing SIMCA 17 software. Antibacterial activities were assessed using the microdilution method on Staphylococcus aureus bacteria. The analysis of antibacterial activity was determined using the Probit analysis method to ascertain the Minimum Inhibitory Concentration (MIC) 50 value. This study also involved the extraction of essential oils from emprit ginger rhizomes. The individual profiles of these essential oils were determined via Gas Chromatography Mass Spectrometry. The essential oil profiles of emprit ginger were subjected to a multivariate calibration using Principal Component Analysis facilitated by the SIMCA 17 software. Antibacterial activity tests were conducted using the microdilution method on Staphylococcus aureus bacteria. The antibacterial activity was ascertained using probit analysis to derive the Minimum Inhibitory Concentration 50 (MIC 50) values. The highest MIC 50 of emprit ginger rhizome on Staphylococcus aureus bacteria was found in samples from Wonogiri, with a concentration of 0.3408%. Compounds displaying significant discriminative influence on Staphylococcus aureus bacteria included Z-Citral, Geranyl acetate, Zingiberenol, Beta-Myrcene, (1S0-2,6,6-Trimethylbicyclo [3.1.1] hept-2-ene, and Bicyclo [2.2.1]heptan-2-ol, 1,7,7-trimethyl, exo-(CAS)

GSH-conjugation Reduces PGV-1 Cytotoxicity and Its Ability in Downregulating N-Myc, β-catenin, and p62 Protein in Huh-6 Cells Utomo, Rohmad Yudi; Meiyanto, Edy; Susidarti, Ratna Asmah
The Indonesian Biomedical Journal Vol 17, No 4 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i4.3634

BACKGROUND: Pentagamuvone-1 (PGV-1), a synthetic curcumin analogue, exhibits potent anticancer activity against Hepatocellular Carcinoma (HCC) by disrupting cell cycle regulation and downregulating key oncogenes such as N-Myc. Numerous studies have examined the role of glutathione (GSH) conjugation in modulating the anticancer properties of curcumin and its analogues. In contrast, the impact of PGV-1 metabolism, particularly GSH conjugation, and its implications for anticancer efficacy have not yet been elucidated. This study was performed to prepare GSH-conjugated PGV-1 (PGV-1-(GSH)2) as the model of PGV-1 metabolite and evaluate its potential distinct cytotoxicity on Huh-6 cells.METHODS: PGV-1 was synthesized via an acid-catalyzed reaction between 4-hydroxy-3,5-dimethylbenzaldehyde and cyclopentanone while PGV-1-(GSH)2 was obtained through reflux at 70oC for 2 hours. The cytotoxic effects of PGV-1 and PGV-1-(GSH)2 on Huh-6 and JHH4, two HCC cells, were assessed using a cell counting kit-8 (CCK-8) assay, while immunoblotting was performed to evaluate their impact on N-Myc and its downstream protein such as β-catenin, and p62.RESULTS: PGV-1-(GSH)2 was prepared through GSH conjugation of PGV-1 in orange color solution, as confirmed by Electrospray Ionization Mass Spectrometry (ESI-MS), Fourier Transform Infrared Spectroscopy (FT-IR), and Nuclear Magnetic Resonance (NMR) analysis. Cytotoxicity assays revealed that PGV-1-(GSH)2 exhibited less potent anticancer activity against HCC cells than PGV-1. GSH conjugation also decreased the ability of PGV-1 in downregulating the N-Myc, β-catenin, and p62 protein level.CONCLUSION: The prepared PGV-1-(GSH)2 reduces the cytotoxicity of PGV-1 and its ability on downregulating N-Myc, β-catenin, and p62 in Huh-6 cells. These findings highlight the need for further exploration about the study of PGV-1 metabolism which could affect the anticancer efficacy against HCC.KEYWORDS: curcumin, PGV-1, GSH, HCC, N-Myc

DPPH Scavenging Activity, Reducing Power, and Metal Chelating Capacity of Compound 1-(2,5-dihydroxyphenyl)-3-pyridine-2-yl-propenone Octa Ujiantari, Navista Sri; Wibowo, Andy Eko; Susidarti, Ratna Asmah
Journal of Food and Pharmaceutical Sciences Vol 11, No 3 (2023): J.Food.Pharm.Sci
Publisher : Integrated Research and Testing Laboratory (LPPT) Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jfps.7317

Background: The excessive free radicals in the body could stimulate oxidative stress which has been implicated in several diseases such as cancer, inflammation, aging, and cardiovascular diseases. The presence of antioxidants can diminish the reactivity of free radicals. The antioxidant defense system in the body may be insufficient thus intake of dietary antioxidants is recommended. Natural flavonoid such as chalcone has been known to exert several biological activities, especially antioxidant activity. Compound 1-(2,5-dihydroxyphenyl)-3-pyridine-2-yl-propenone was successfully synthesized using microwave radiation and several studies reported its biological activities such as antiinflammation, anticancer, and antibacterial activities. Those activities are thought to be related to antioxidant mechanisms. Therefore, this study aimed to evaluate its antioxidant activity. Methods: The antioxidant activities were performed with three different methods: DPPH scavenging activity, ferric reducing power, and metal chelating capacity. Results: 1-(2,5-dihydroxyphenyl)-3-pyridine-2-yl-propenone showed DPPH scavenging activity, ferric reducing power, and metal chelating capacity with IC50S values of 4,471 ± 0,052 μg/mL; 156.56 ± 4.42μg/mL; and 6,273 ± 0,025 μg/mL, respectively. Conclusions: Our results found that 1-(2,5-dihydroxyphenyl)-3-pyridine-2-yl-propenone had quite potent antioxidant activity.

The Doxorubicin-Induced G2/M Arrest in Breast Cancer Cells Modulated by Natural Compounds Naringenin and Hesperidin Junedi, Sendy; Hermawan, Adam; Fitriasari, Aditya; Setiawati, Agustina; Susidarti, Ratna Asmah; Meiyanto, Edy
Indonesian Journal of Cancer Chemoprevention Vol 12, No 2 (2021)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev12iss2pp83-89

Doxorubicin as the common drug for breast cancer has been widely proposed to use in combine with a natural compound in order to overcome its side effects such as cardiotoxicity and resistance. Previously, we reported that naringenin and hesperidin, the abundant flavanons in citrus fruit peel, increased cytotoxic and apoptosis activities of doxorubicin in doxorubicin resistant breast cancer cells (T47D and MCF-7 cells). Since doxorubicin arrests G2/M phase in most cancer cells, both flavanons are speculated to affect the similar phase in breast cancer cells. Cell cycle distributions were determined by flowcytometry using propidium iodide (PI) to stain DNA of the cells. Combination of naringenin or hesperidin with doxorubicin increased accumulation of T47D cells in G2/M phase, while in MCF-7 cells, accumulated cells in G2/M phase were decreased, accompanying with slightly increased in G1 phase. Naringenin itself had no effect on cell cycle of both cells. Whereas, hesperidin arrested G2/M and G1 phases in T47D and MCF-7 cells, respectively. The different effect of naringenin and hesperidin in T47D and MCF-7 cells is most likely caused by difference of p53 status. In p53 mutant, T47D cells, naringenin and hesperidin supported mechanism of doxorubicin to arrest at G2/M that to be considered via p53-independent pathway. Whereas, in p53 wild-type MCF-7 cells, naringenin and hesperidin decreased G2/M arrest, suggesting that both flavanons do not utilize cell cycle arrest for their anticancer activity with doxorucibin. This study revealed that potential co-chemoterapeutic agents, naringenin and hesperidin distinctly modulated cell cycle arrest induced by doxorubicin according to the characteristic of breast cancer cells.Keywords: naringenin, hesperidin, doxorubicin, cell cycle, breast cancer cells.

Growth Inhibitory Property of Pentagamavunone-0 (PGV-0) on 4T1 Cells under Stress Condition : 2D and 3D Culture Model Muflikhasari, Haruma Anggraini; Jenie, Riris Istighfari; Susidarti, Ratna Asmah; Meiyanto, Edy
Indonesian Journal of Cancer Chemoprevention Vol 10, No 3 (2019)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev10iss3pp149-158

Pentagamavunone-0 (PGV-0), one of the curcumin analogue, is reported to have a cytotoxic effect on various cancer cells. This study aimed to explore the growth inhibitory effects of PGV-0 against highly-metastatic breast cancer, 4T1 cells under stress condition covering 2D and 3D speroid cytotoxic, anti-migration, and suppression of MMP-9. PGV-0 showed cytotoxic effects on 2D and 3D 4T1 cells with IC50value of 49 μM and 26 μM, respectively. In addition, PGV-0 performed anti-migratory effect. The single treatment at 25 μM PGV-0 and 50 μM showed inhibitory effect on cell migration by 54% and 51% respectively. whilst, the combination of PGV-0 at the concentration of 25 μM and 50 μM with doxorubicin significantly inhibited cell migration by 41% and 38%, respectively. The gelatin zymography assay showed that PGV-0 decreased MMP-9 expression both in a single treatment and in combination with doxorubicin. In conclusion, PGV-0 is potential to be developed as anti-tumorigenesis agent on highly-metastatic breast cancers.Keywords: Pentagamavunone-0 (PGV-0), anti-migration, MMP-9, 4T1 cells, spheroid

Citrus sinensis Peel Extract Synergistically Enhances the Cytotoxic Effect of Chemotherapeutic Agents on HepG2 Cells Zufairo, Shofa Khamdanatuz; Rahmawati, Desty Restia; Meiyanto, Edy; Susidarti, Ratna Asmah
Indonesian Journal of Cancer Chemoprevention Vol 14, No 3 (2023)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev14iss3pp151-159

Doxorubicin (DOX) and cisplatin (Cis), non-specific chemotherapeutic agents used for hepatocellular carcinoma (HCC), are frequently combined with synthetic or natural agents to enhance their cytotoxic effects. Citrus sinensis peel extract (CPE) serves as a natural source of flavonoids, including sinensetin (SIN), which has the potential to increase the efficacy of DOX and Cis. This study aimed to observe the effect of CPE and SIN one of CPE compounds, in enhancing liver cancer cell susceptibility to doxorubicin and cisplatin. The assays conducted in this study included a phytochemical analysis of CPE using TLC, cell viability assays against HepG2 cells using MTT assay in both single and combination forms, and cell viability assays on Vero cells. The result confirmed the presence of SIN as one of the compounds in CPE. Both CPE and SIN, when used individually, exhibited moderate cytotoxic effects on HepG2 cells with IC50 of 101.09 μg/mL and 83.13 μM, respectively, while showing no cytotoxic effect on Vero cells. Cis demonstrated significant cytotoxicity against HepG2 cells with an IC50 of 7.86 μM. DOX exerted a strong cytotoxic effect on both HepG2 and Vero cells, with the IC50 of 2.52 μM and 13.98 μM. It was observed that CPE was able to synergistically enhance the cytotoxic effects of DOX, and SIN synergistically increased the cytotoxicity of Cis, particularly against HepG2 cells, with CI<1.0.Keywords: CPE, SIN, Cisplatin, Doxorubicin, HCC.

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