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All Journal Indonesian Journal of Cancer Chemoprevention Journal of Mathematical and Fundamental Sciences Indonesian Journal of Biotechnology Majalah Obat Tradisional Journal of Tropical Biodiversity and Biotechnology INDONESIAN JOURNAL OF PHARMACY The Indonesian Biomedical Journal JURNAL MANAJEMEN DAN PELAYANAN FARMASI (Journal of Management and Pharmacy Practice) JPSCR : Journal of Pharmaceutical Science and Clinical Research JFIOnline Jurnal Ilmu Kefarmasian Indonesia Majalah Kesehatan Pharmamedika
Muthi Ikawati
Faculty Of Pharmacy, Universitas Gadjah Mada
Agriculture, Biological Sciences & Forestry Astronomy Biochemistry, Genetics & Molecular Biology Chemistry Earth & Planetary Sciences Environmental Science Health Professions Immunology & microbiology Materials Science & Nanotechnology Mathematics Medicine & Pharmacology Physics Public Health

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Immunomodulatory Effect of Dioscorea esculenta L. on NF-κB, TLR-4, TNF-α, and IL-10 Expressions in LPS-stimulated RAW 264.7 Mouse Macrophages Puspitaningrum, Ika; Ikawati, Muthi; Fakhrudin, Nanang; Nurrochmad, Arief
The Indonesian Biomedical Journal Vol 17, No 3 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i3.3630

Abstract

BACKGROUND: Gene expressions of toll-like receptor 4 (TLR)-4, nuclear factor-kappaB (NF-κB), tumor necrosis factor (TNF)-α, and interleukin (IL)-10 are known to have roles in the inflammatory process and affect the regulation of the immune system. A preliminary study showed that Dioscorea esculenta L. tuber has immunomodulatory activity against macrophage phagocytosis activity and lymphocyte proliferation. However, the immunomodulatory activity of aqueous extract (AE), polysaccharide fraction (PF), and non-polysaccharide fraction (NPF) of D. esculenta L. tubers on these gene expressions have not been elucidated well. Therefore, this study was performed to determine its immunomodulatory activity by utilizing RAW 264.7 cell culture induced by lipopolysaccharide (LPS).METHODS: RAW 264.7 cells were stimulated with LPS at a concentration of 1 µg/mL for 30 minutes before incubation with non-toxic concentrations of AE, PF, NPF, positive control, and inulin at 25 and 50 µg/mL. TNF-α, IL-10, TLR-4, NF-κB, and β-actin expressions were evaluated using reverse transcription-polymerase chain reaction (RT-PCR) and were normalized with β-actin as an internal control. Triplicate experiments were performed throughout this study.RESULTS: Treatment with 25 µg/mL NPF significantly decreased the expression of NF-κB, TLR-4, and TNF-α (p<0.05). In contrast, treatment of 25 and 50 µg/mL PF significantly decreased the NF-κB expression (p<0.05). Moreover, only treatment with 50 µg/mL AE exhibited a significant increase in IL-10 expression (p<0.05).CONCLUSION: Treatment with D. esculenta L. tuber stimulated macrophage RAW 264.7 cells via NF-κB, TLR-4, TNF-α, and IL-10 expressions. NPF at 25 µg/mL has stronger immunomodulatory activity in reducing the expression of genes involved in the inflammatory process that plays a role in regulating the immune system.KEYWORDS: Dioscorea esculenta L., Immunomodulator, IL-10, NF-κB, TLR-4, TNF-α, RAW 264.7 cell
Diosmin Enhances the Anti-migration Activity of Curcumin Analog PGV-1 on Colorectal Cancer Cells Ikawati, Muthi; Utomo, Rohmad Yudi; Hapsari, Novia Permata; Meiyanto, Edy; Oka, Chio
The Indonesian Biomedical Journal Vol 16, No 1 (2024)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v16i1.2829

Abstract

BACKGROUND: Diosmin enhances the cytotoxicity of Pentagamavunone-1 (PGV-1) in cancer cells. PGV-1 and diosmin are predicted to target several matrix metalloproteinases (MMPs) in metastatic cancer, including colorectal cancer, but the anti-migration potency of their combination has not established yet. This study evaluates the anti-migration effect of PGV-1 and diosmin combination in colorectal cancer.METHODS: The cytotoxicity assay using Cell Counting Kit 8 (CCK-8) method in WiDr colorectal cancer cells was carried out to determine the concentration for anti-migration experiments. The wound healing assay was used to observe the anti-migration activity by measuring the cell-free area. Gelatin zymography was employed to detect the MMP activity indicating by the clear band density. The interaction between PGV-1 or diosmin and MMP proteins was predicted by molecular dockings.RESULTS: PGV-1 was cytotoxic (IC50 17 mM), while diosmin up to 100 mM did not affect cell viability. Both 10 mM PGV-1 as well as 50 and 100 mM diosmin slowed down the closure of cell-free area. A 100 mM diosmin was significantly enhance the anti-migratory activity of 50 and 100 mM PGV-1. The activity of MMP-9 and MMP-2 was also lower in the presence of diosmin compared to than that of PGV-1 alone. PGV-1 or diosmin was also able to interact with MMP proteins with a lower energy compared to than that of the native ligands.CONCLUSION: Diosmin enhances the anti-migration activity of PGV-1 in WiDr cells, possibly by affecting MMPs’ activity. This study is an evidence that diosmin is a potential co-chemotherapy candidate for PGV-1, that can be utilized to overcome metastatis in colorectal cancer.KEYWORDS: cancer, citrus flavonoid, co-chemotherapy, diosmin, matrix metalloproteinases (MMPs), migration, Pentagamavunone-1, WiDr cancer cell
Phytochemical and Bioinformatic Studies of Citrus Flavonoids as Chemopreventive Agents Targeting GGPS1 for Liver Cancer Wardani, Ratih Kurnia; Rhamandana, I Made; Gono, Christina Mutiara Putri; Ikawati, Muthi
Indonesian Journal of Cancer Chemoprevention Vol 12, No 3 (2021)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev12iss3pp137-147

Abstract

Overexpression of geranylgeranyl diphosphate synthase 1 (GGPS1) is an unfavorable prognosis in liver cancer development. The side effects of therapeutic standards encourage the development of therapeutic agents from herbal materials. Citrus peels are rich of phytochemical compounds, especially citrus flavonoids, that possess cytotoxic activities. This study aimed to determine the potential of citrus flavonoids as chemopreventive agents targeting GGPS1 protein by phytochemical and bioinformatic studies. Dried peels of Citrus reticulata were extracted by hydrodynamic-cavitation method followed by identification of compounds using thin layer chromatography (TLC). The expression level of GGPS1 was obtained from UALCAN, while its correlation with survival rate was obtained from the GEPIA. Prediction models regarding the potential inhibitors of citrus peel compounds against GGPS1 were obtained through KNIME and ChEMBl, followed by literature studies on chemopreventive activity of citrus flavonoids. The molecular docking was used to predict the molecular interaction followed by tracking of target genes that were positively correlated with GGPS1 by SwissTargetPrediction. Yielded 75% (v/v), the extract positively contained citrus flavonoid with hesperidin as comparison. Overexpression of GGPS1 significantly reduced the survival rate of liver cancer patients (p value=0.019). Four citrus flavonoid compounds, namely tangeretin, nobiletin, hesperidin, and naringenin showed potential inhibition to GGPS1. The molecular docking showed that tangeretin had a strong affinity compared to the native ligand and zoledronic acid, as positive control. PARP1, CSNK2A1, TNKS2, and GSK3B were clarified as targeted genes for tangeretin and nobiletin that positively correlated with GPPS1. In vitro and in vivo studies will validate our findings and support the development of citrus peel extract with rich flavonoid contents as a chemopreventive agent.Keywords: geranylgeranyl diphosphate synthase 1 (GGPS1), liver cancer, hydrodynamic-cavitation, citrus flavonoid, bioinformatic.
Chromolaena odorata L. Leaf Extract Elevates Cytotoxicity of Doxorubicin on 4T1 Breast Cancer Cells Putri, Amaliya Permata; Rahmawati, Desty Restia; Rahman, Faaza Aulia; Meiyanto, Edy; Ikawati, Muthi
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/indonesianjcanchemoprev14iss3pp160-170

Abstract

Chemotherapeutic agents for breast cancer such as doxorubicin can attack normal cells as the side effects. Chromolaena odorata L. and its chemical content, sinensetin, have potential anticancer  and  antioxidant  properties.  The  objective  of  this  research  is  to  examine  the anticancer properties of C. odorata leaves extract and sinensetin on 4T1 triple negative breast cancer (TNBC) cells combined with doxorubicin. The MTT (3-(4, 5-dimethylthiazolyl-2)-2, 5 diphenyltetrazolium  bromide)  assay  on  4T1  cells  was  used  to  determine  the  IC50 and the Combination  Index  (CI)  of  the  two  agents  in  combination.  Washing  out the  treatment  and determining  the  cells  viability  after  a  few  days  was done  to evaluate  the  persistence  of the  effects  to  cancer  cells.  Chromolaena odorata  extract  (COE)  obtained  was  proven  to contain  sinensetin  which  gave  a positive  signal  on  the  chromatogram.  COE  and  sinensetin were  moderately  cytotoxic  to  4T1  cells  with  IC50  value  of  53  μg/mL  and  58  μM  (21.6 μg/mL), respectively. Both compounds were synergist (CI<0.7) to strong synergist (CI<0.3) when combined with doxorubicin (IC50 90 nM = 0.05 μg/mL). COE and sinensetin exhibited moderate and not cytotoxic against Vero cells with IC50 values of 60 μg/mL and 243 μM (90.43 μg/mL), respectively. Both COE and sinensetin showed selectivity index values of >1 (1.13 and 4.19, respectively).  Moreover,  the  cytotoxic  effects  of  COE  on  4T1  cells  was  persisted  until  48  h after  removing  COE  from  the  medium,  indicating  the  tumor-suppression  potency  of  COE. Our findings strengthen the scientific basis of C. odorata leaves extract to be developed as a co-chemotherapeutics agent for doxorubicin on TNBC.Keywords: Chromolaena odorata L., breast cancer cells, doxorubicin, co-chemotherapy, kidney cells.
Citrus Flavonoids from Citrus reticulata Peels Potentially Target an Autophagy Modulator, MAP1LC3A, in Breast Cancer Anggoro, Bayu; Kumara, Dennaya; Angelina, Dhella; Ikawati, Muthi
Indonesian Journal of Cancer Chemoprevention Vol 12, No 3 (2021)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev12iss3pp114-122

Abstract

Citrus flavonoids have been known for their vast biological activities including chemoprevention activities. However, the organic solvent extraction system limits its potential utilization. We recently adopted a hydrodynamic-cavitation method to extract citrus flavonoids from citrus peels. In this study we verified the high flavonoid content of the hydrodynamic-cavitation extract from Citrus reticulata peels and explore the potency of its citrus flavonoid contents as targeted chemoprevention agent for breast cancer by using bioinformatics. Based on a thin layer chromatography, the extract positively yielded high content of citrus flavonoids represented by hesperidin. The toxicity analysis by Protox II Online Tool revealed that hesperidin as the major citrus flavonoid in the extract was considered safe with a predicted LD50 of 12,000 mg/kg. We then further exploring citrus flavonoids’ capacity in targeting MAP1LC3A, a key protein in autophagy. UALCAN analysis validated that low expression of MAP1LC3A is associated with low survival rates in breast cancer patients. Limonin, hesperidin, narirutin, neohesperidine, and naringin are flavonoids from citrus peels that predicted to have inhibitory activity against Protein Kinase A (PKA), a negative upstream of MAP1LC3A, calculated by KNIME. Citrus flavonoids scoparone, cirsimaritin, 4',5,7-trimethoxyflavone, eupatorine, and hesperidin were also exhibit similar structure to an agonist of ATG4B, a protein that plays a role in MAP1LC3A activation. Furthermore, eupatorine, hesperidin, and cirsimaritin displayed a high affinity to ATG4B based on a molecular docking. We concluded that citrus flavonoids from citrus peels are safe to normal cells, and the citrus flavonoids potentially targets MAP1LC3A by inhibiting PKA and acting as ATG4B agonists. Thus, this extract-contained flavonoids from citrus peels is potential to be investigated further as a chemoprevention agent by inducing autophagy, especially for breast cancer.Keywords: Citrus reticulata, citrus flavonoid, autophagy, MAP1LC3A, breast cancer.
Co-Authors . Anindyajati Abdul Manaf Ali, Abdul Manaf Adam Hermawan ADTYA FITRIASARI Afifah, Anis Afivah Dewi Anggraeni Agusta Fauzi, Ilham Alfi Yasmina Angelina, Dhella Anggoro, Bayu ANINDYAJATI ANINDYAJATI Anindyajati Anindyajati ANNISA KARAMITA Annisa Khumaira Arief Nurrochmad Astrid Ayu Maruti Astrid Ayu Maruti Chio Oka, Chio Dewi Pratiwi Dewi Pratiwi Dhania Novitasari Dhiya Ulhaq Salsabila Dyaningtyas D.P. Putri Dyaningtyas Dewi Putri Pamungkas EDIATI EDIATI Ediati Sasmito Edy Meiyanto Edy Meiyanto Edy Meiyanto Edy Meiyanto Edy Meiyanto EDY MEIYANTO Edy Meiyanto Edy Meiyanto Edy Meiyanto Edy Meiyanto Edy Meiyanto Een Sri Endah Endah Puji Septisetyani Endah Puji Septisetyani, Endah Puji Erlina Rivanti Erlina Rivanti Febri Wulandari Fikri Amalia Gono, Christina Mutiara Putri Hanifa, Mila Hapsari, Novia Permata Hilyatul Fadliyah Ibrahim Arifin Ika Puspitaningrum Ika Putri Nurhayati Ilham Augusta F Ilham Augusta F. Imaniyyati, Niar Nurul Inna Armandari Inna Armandari Jenie, Riris Istighfari Jun-Ya Kato Jun-ya Kato Kumara, Dennaya Mintarsih, Betty Mintarsih, Betty Mitsunori Kirihata Moordiani Moordiani Muhammad Novrizal Abdi Sahid Mustofa Mustofa Nanang Fakhrudin, Nanang Nanda Resa Pratama Nanda Resa Pratama Natasia, Nyssa Niken Nur W Niken Nur W, Niken Nindya Budiana Putri Normaidah, Normaidah Novi Hastuti Novi Hastuti, Novi Nurma Sabila Nurramadhani A. Sida Purwanto, Heri Putri, Amaliya Permata Rahman, Faaza Aulia Rahmani, Mawardi Rahmani, Mawardi Rahmawati, Desty Restia RATIH HARDIKA PRATAMA Ratih Hardika Pratama Ratih Hardika Pratama Ratna Asmah Susidarti Ratna Asmah Susidarti Ratna Asmah Susidarti Rhamandana, I Made Rifai, Fauziah Novita Putri RIRIS ISTIGHFARI JENIE Rita Riata Rita Riata Ritmaleni, Ritmaleni Rohmad Yudi Utomo Sagiyo, Marrita Langgeng Sari Haryanti Sari Haryanti Shirly Kumala Sismindari, Sismindari Sri Handayani Sri Kasianningsih Sri Kasianningsih Sri Pudjiraharti Sukari, Mohd. Aspollah Sukari, Mohd. Aspollah Susi Ari Kristina Susi Ari Kristina Susi Ari Kristina Syifa Athia Zainun Faqiha Tafrihani, Ahmad Syauqy Wardani, Ratih Kurnia Wulandari Wulandari Yogi Ertanto Yogi Ertanto Yohanes, Jasson Yurista Gilang Yurista Gilang YURISTA GILANG IKHTIARSYAH Ziana Walidah Zulfin, Ummi Maryam