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Hesperidin Increases Cytotoxic Activity of Doxorubicin on Hela Cell Line Through Cell Cycle Modulation and Apoptotis Induction Kusharyanti, Indri; Larasati, .; Susidarti, Ratna Asmah; Meiyanto, Edy
Indonesian Journal of Cancer Chemoprevention Vol 2, No 2 (2011)
Publisher : Indonesian Research Gateway

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Abstract

Combination of chemotherapeutic agent and chemo preventiveagent is being a new approach in cancer treatment.This is aimed at enhancing the effectivity and also reducing drugresistance and adverse side effect of the chemo therapeuticagent.Hesperidin,acitrus flavonoid has reported to reduce theproliferation of many cancer cells.The objectives of this study were to investigate cytotoxic activities, cell cycle modulation and apoptosis induction of he speridinand its combination withdoxorubicinon Helacelllines.MTT [3-(4,5-dimethylthiazol-2-yl)-2.5-diphenyltetrazoliumbromide] assay was used tomeasure the growth inhibitory effect of he speridinanditscombination with doxorubicinon Helacells.Cellcycle profile was determined by flowcytometry and the dataobtained was analyzed by using Mod Fit LT3.0program.Apoptos is assay was done using double staining method usingethidium$bromideandacridine$orange.Hesperidin inhibited cellgrowth with IC5048M, while the IC50 of doxorubicin was 1000nM.Combination of 500n Mdoxorubicin and 6M hesperidin showed strongest inhibitory effect toward Hela cells. Hesperidin of 24 2M accumulated HeLacells at G1phase,butit scombinationwith 500nM Doxorubicin gave G1 and Sphase accumulation at 24h incubation.Both of Hesperidin and Doxorubicin were capable of inducing apoptosis.Inaccordance of the apoptoticeffect,hesperidin,doxorubicin and their combination decreasedthe expression Bcl$2 and increased the expression of Bax. Accordingtothisresult,hesperidinhasapotencytobedevelopedasco$chemotherapeutic agent forcervical cancer. Key    words:Cochemotherapy,Hesperidin,Doxorubicin,Hela,MTTassay
Naringenin Enhances the Anti-Tumor Effect of Doxorubicin on HeLa Cervical Cancer Cells Through Cytotoxic Activity and Apoptosis Induction Larasati, .; Kusharyanti, Indri; Hermawan, Adam; Susidarti, Ratna Asmah; Meiyanto, Edy
Indonesian Journal of Cancer Chemoprevention Vol 2, No 3 (2011)
Publisher : Indonesian Research Gateway

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Abstract

Naringenin, an abundant flavanon in the peel of citrus fruits is reported to possess anti-proliferative  effect  in  many  cancer  cells.  Herein,  we  investigated  the  cytotoxic  effect  and apoptosis  induction  of  naringenin  in  combination  with  doxorubicin  on  HeLa  cells.  The cytotoxicity assay of naringenin, doxorubicin, and their combination were carried out by using MTT  assay.  Cell  viability  was  used  as  the  parameters  to  evaluate  combination  effectiveness. Cell  cycle  distribution  was  determined  by  flow  cytometry  and  analyzed  using  ModFit  LT  3.0 program.  Apoptosic  assay  was  done  by  double  staining  method  using  Ethidium  Bromide-Acridine  Orange.  Investigation  on  the  expression  of  Bax  and  Bcl-2  were  determined  by immunocytochemistry method. Naringenin and doxorubicin showed cytotoxic effect  on HeLa cells  with  their  IC50  values  of  195  µM  and  1  µM,  respectively.  Whereas  combination  of naringenin  -  doxorubicin  showed  greater  cytotoxicity  compared  the  single  treatment  of doxorubicin.  The  strongest  cytotoxic  activity  was  observed  at  a  combination  of  100  µM naringenin  and  0,5  µM  doxorubicin.  Single  treatment  of  0,5  µM  doxorubicin  for  24  hours  on HeLa cells induced  S-phase arrest while 100 µM naringenin did not affect on HeLa cell cycle. The  combination  induced  S-phase  arrest  with  the  increased  of  sub-G1  phase  percentage.  In accordance with the flow cytometry results, the double staining apoptosis assay results showed the increase of apoptotic cells. Naringenin, doxorubicin, and their combination also increased the  expression  of  Bax  and  decreased  the  expression  of  Bcl-2.  These  results  concluded  that naringenin was a potential co-chemotherapy agent for cervical cancer due to its synergism with doxorubicin.Keywords:  co-chemotherapy,  naringenin,  doxorubicin,  HeLa  cells,  cytotoxicity,  cell  cycle, apoptosis
UJI TOKSISITAS AKUT FRAKSI N-HEKSAN EKSTRAK METANOL DAUN DAN BATANG Impatiens balsamina Linn. DENGAN PEDOMAN OECD 425 Sunggono, Benny Wijaya; Kusharyanti, Indri; Nurbaeti, Siti Nani
Majalah Obat Tradisional Vol 19, No 3 (2014)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (954.094 KB) | DOI: 10.14499/mot-TradMedJ19iss3pp118-126

Abstract

Tanaman pacar air (Impatiens balsamina Linn.) merupakan tanaman herba yang secara empirik digunakan untuk terapi nyeri tulang, gigitan serangga, peluruh haid dan pencegah kanker pencernaan. Secara empiris, daun pacar air tidak boleh dikosumsi secara langsung karena mengandung racun yang dapat mempengaruhi pencernaan. Tujuan penelitian ini adalah untuk menentukan tingkat keamanan dari fraksi n-heksan daun dan batang pacar air sehingga dapat dijadikan acuan untuk penggunaan terapi yang aman. Fraksi n-heksan diperoleh dari ekstraksi cair cair ekstrak metanol daun dan batang pacar air. Hasil penapisan fitokimia dengan uji tabung menyatakan bahwa fraksi n-heksan mengandung senyawa triterpenoid steroid. Uji toksisitas akut dilakukan dengan metode yang diadopsi dari OECD (Organization for economic co-operation and development) nomor 425: Acute Oral Toxicity (Up and Down Procedure). Hasil pengujian Limit Test menyatakan fraksi n-heksan daun dan batang pacar air memiliki nilai LD50 lebih besar dari 5000 mg/kg berat badan tikus. Hingga dosis 5000 mg//kgBB tidak ada tanda-tanda toksik pada sistem mata, respirasi, kelakuan, otonom dan somatomor. Pada dosis 5000 mg/kgBB, fraksi n-heksan juga tidak menyebabkan kematian pada tikus, tidak menyebabkan penurunan berat badan, tidak menyebabkan perubahan makan dan minum yang signifikan. Pemberian fraksi n-heksan pada dosis 2000 mg/kg dapat menyebabkan lesi degenarasi hidropik pada organ hati dan dapat memperparah kerusakan pada organ ginjal pada dosis 5000 mg/kg. Oleh sebab itu, penggunaannya tetap harus diperhatikan. Menurut klasifikasi Loomis, fraksi n-heksan pacar air berada dalam kategori toksik ringan.
AKTIVITAS ANTIMAKAN EKSTRAK KULIT BATANG MUNDU (Garcinia dulcis (Roxb.) Kurz) TERHADAP LARVA Epilachna varivestis Widiyantoro, Ari; ., Kustiati; Kusharyanti, Indri
Sainstek Vol 3, No 3, 2008
Publisher : Jurnal Sainstek

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Abstract

Isolasi senyawa antimakan dari 1,5 kg kulit batang mundu (Garcinia dulcis (Roxb.) Kurz) menghasilkan senyawa golongan santon (50 mg) berupa kristal berwarna kuning dengan titik leleh 208210oC dan uji fitokimia menunjukkan positif polifenol. Spektrum UV isolat menunjukkan adanya gugus kromofor seperti gugus C=O, C=C aromatik, dan OH. Spektrum IR isolat menunjukkan adanya gugus fungsi seperti OH, C=O, C-O-C, C=C aromatik, dan C-H aromatik.Uji aktivitas antimakan terhadap Epilachna varivestis menunjukkan bahwa fraksi etil asetat batang mundu pada konsentrasi 0,5% tidak menunjukkan aktivitas antimakan. Sementara pada konsentrasi 2% menunjukkan rata-rata ratio antimakan 50,78 4,87 terhadap placebo. Aktivitas antimakan ini disebabkan oleh aktivitas senyawa yang terdapat dapat fraksi etil asetat yaitu golongan senyawa polifenol dan flavonoid. Aktivitas antimakan isolat (positif polifenol) kulit batang mundu terhadap Epilachna varivestis pada konsentrasi 0,5 % menunjukkan rata-rata ratio antimakan terhadap placebo sebesar 35,45 4,51 Sementara pada konsentrasi 2% menunjukkan rata-rata ratio antimakan 91,87 4,56 terhadap placebo.
Senyawa Aktif Antiinflamasi dari Kulit Batang Pasak Bumi (Eurycoma longifolia Jack) Ari Widiyantoro; Indri Kusharyanti; Lia Destiarti; Elvi Rusmiyanto Pancaning Wardoyo
EKSAKTA: Journal of Sciences and Data Analysis VOLUME 12, ISSUE 2, August 2011
Publisher : Fakultas Matematika dan Ilmu Pengetahuan Alam

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Abstract

Pasak bumi (Eurycoma longifolia Jack) merupakan salah satu famili Simaroubaceae yang mengandung senyawa kimia dengan aktivitas biologik yang potensial.Penggunaannya di masyarakat selama ini hanya terbatas sebagai aprodisiaka, namun beberapa penelitian menunjukkan ditemukannya senyawa kimia dengan aktivitas sebagai antimalaria, antikanker dan antioksidan. Penelitian ini mengungkapkan adanya senyawa kimia yang bersifat antiinflamasi dengan tinjauan reduksi radang pada kaki tikus yang terinduksi karagenan. Hasil penelitian diperoleh suatu senyawa quassinoid yang bersifat antiinflamasi yaitu eurycomaosida, namun aktivitasnya lebih rendah dibanding indometasin sebagai kontrol positif.   Kata Kunci : Eurycoma longifolia Jack, antiinflamasi, udema, karagenan, quassinoid
UJI TOKSISITAS AKUT FRAKSI N-HEKSAN EKSTRAK METANOL DAUN DAN BATANG Impatiens balsamina Linn. DENGAN PEDOMAN OECD 425 Benny Wijaya Sunggono; Indri Kusharyanti; Siti Nani Nurbaeti
Majalah Obat Tradisional Vol 19, No 3 (2014)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (954.094 KB) | DOI: 10.22146/tradmedj.8228

Abstract

Garden balsam (Impatiens balsamina Linn) is a plant that has been used for joint pain, insect bite, promotes regular menstrual cycle and prevents stomach cancer. Traditionally, the leaves of garden balsam are suspected to contain poison that can affect the digestive system. The purpose of this research is to evaluate the safety of the garden balsam stem and leaf n-hexane fraction so it can be used safely. Stem and leaf Methanol extract of garden balsam was fractioned into n-hexane fraction with liquid extraction method. Phytochemical screening showed that the fraction contains triterpenoid steroid compound. The OECD Guideline no 425: Acute Oral Toxicity (Up and Down Procedure) was adopted to evaluate the safety of the fraction. Two until three months old female Sprague Dawley rat were subjected to n-hexane fraction suspension. Limit test showed that the n-hexane fraction LD50 is greater than 5000 mg/kgBW. There were no clinical sign of toxicity on the eye, respiration system, behavior, autonomic and somatomotoric system up to dose 5000 mg/kgBW. At that dose, the fraction did not cause mortality on rats, and did not lower the body weight, food and water consumption of rats. The fraction also did not cause any histology change to the liver and renal. Our conclusion is the fraction is safe to consume below 5000mg/kgBW. According to Loomis classification, n-hexane fraction of garden balsam steam and leaf has low toxicity.
EFEKTIVITAS KRIM ANTIJERAWAT EKSTRAK METANOL DAUN PACAR AIR (Impatiens balsamina L.) Abdurraafi' Maududi Dermawan; Liza Pratiwi; Indri Kusharyanti
Majalah Obat Tradisional Vol 20, No 3 (2015)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (513.143 KB) | DOI: 10.22146/tradmedj.8851

Abstract

Many studies have found that Impatiens balsamina leaves contain antibacterial activity. The present study was conducted to evaluate antiacne effect from cream of methanolic extract from leaves of Impatiens balsamina L. towards Propionibacterium acnes and Staphylococcus epidermidis, which concentration that gives the most effective antiacne and to evaluate physical and chemical properties of creams. The simplicia was extracted by soxhlet technique and using methanol as solvent. Kirby-Baeur diffusion method was used to measure the antibacterial activity. The extract was formulated in cream with concentration of 10%, 15% and 20%. Evaluation of physical and chemical properties was done by organoleptic testing, spreadability, adhesive force, pH and antibacterial effectiveness. Data was analyzed using R-Commander program 2.14.1 Version. Formula II which contained 15% extract showed the most effective inhibiton to the growth of bacteria. The evaluation of FII cream showed that the cream was homogen, the spreadability was 21,24 + 1,464cm2, adhesive force more than 60 minutes, and pH of 6,53 + 0,057.
Hesperidin Increase Cytotoxic Activity of Doxorubicin on Hela Cell Line Through Cell Cycle Modulation and Apoptotis Induction Indri Kusharyanti; Larasati Larasati; Ratna Asmah Susidarti; Edy Meiyanto
Indonesian Journal of Cancer Chemoprevention Vol 2, No 2 (2011)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev2iss2pp267-273

Abstract

Combination of chemotherapeutic agent and chemopreventive agent is being a new approach in cancer treatment. This is aimed at enhancing the effectivity and also reducing drug resistance and adverse side effect of the chemotherapeutic agent. Hesperidin, a citrus flavonoid has reported to reduce the proliferation of many cancer cells. The objectives of this study were to investigate cytotoxic activities, cell cycle modulation and apoptosis induction of hesperidin and its combination with doxorubicin on Hela cell lines. MTT [3-(4,5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide] assay was used to measure the growth inhibitory effect of hesperidin and its combination with doxorubicin on Hela cells. Cell cycle profile was determined by flowcytometry and the data obtained was analyzed by using ModFit LT 3.0 program. Apoptosis assay was done using double staining method using ethidium-bromide and acridine-orange. Hesperidin inhibited cell growth with IC50 48 μM, while the IC50 of doxorubicin was 1000 nM. Combination of 500 nM doxorubicin and 6 μM hesperidin showed strongest inhibitory effect toward Hela cells. Hesperidin of 24 µM accumulated HeLa cells at G1 phase, but its combination with 500 nM Doxorubicin gave G1 and S phase accumulation at 24 h incubation. Both of Hesperidin and Doxorubicin were capable of inducing apoptosis. In accordance of the apoptotic effect, hesperidin, doxorubicin and their combination decreased the expression Bcl-2 and increased the expression of Bax. According to this result, hesperidin has a potency to be developed as co-chemotherapeutic agent for cervical cancer.Keywords: Cochemotherapy, Hesperidin, Doxorubicin, Hela, MTT assay
Naringenin Enhances the Anti-Tumor Effect of Doxorubicin on HeLa Cervical Cancer Cells Through Cytotoxic Activity and Apoptosis Induction Larasati Larasati; Indri Kusharyanti; Adam Hermawan; Ratna Asmah Susidarti; Edy Meiyanto
Indonesian Journal of Cancer Chemoprevention Vol 2, No 3 (2011)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev2iss3pp325-333

Abstract

Naringenin, an abundant flavanon in the peel of citrus fruits is reported to possess anti-proliferative effect in many cancer cells. Herein, we investigated the cytotoxic effect and apoptosis induction of naringenin in combination with doxorubicin on HeLa cells. The cytotoxicity assay of naringenin, doxorubicin, and their combination were carried out by using MTT assay. Cell viability was used as the parameters to evaluate combination effectiveness. Cell cycle distribution was determined by flow cytometry and analyzed using ModFit LT 3.0 program. Apoptosic assay was done by double staining method using Ethidium Bromide-Acridine Orange. Investigation on the expression of Bax and Bcl-2 were determined by immunocytochemistry method. Naringenin and doxorubicin showed cytotoxic effect on HeLa cells with their IC50 values of 195 µM and 1 µM, respectively. Whereas combination of naringenin - doxorubicin showed greater cytotoxicity compared the single treatment of doxorubicin. The strongest cytotoxic activity was observed at a combination of 100 µM naringenin and 0,5 µM doxorubicin. Single treatment of 0,5 µM doxorubicin for 24 hours on HeLa cells induced S-phase arrest while 100 µM naringenin did not affect on HeLa cell cycle. The combination induced S-phase arrest with the increased of sub-G1 phase percentage. In accordance with the flow cytometry results, the double staining apoptosis assay results showed the increase of apoptotic cells. Naringenin, doxorubicin, and their combination also increased the expression of Bax and decreased the expression of Bcl-2. These results concluded that naringenin was a potential co-chemotherapy agent for cervical cancer due to its synergism with doxorubicin.Keywords: co-chemotherapy, naringenin, doxorubicin, HeLa cells, cytotoxicity, cell cycle, apoptosis
Formulasi Gel Pacar Air (Impatiens balsamina Linn.) terhadap Propionibacterium acnes dan Staphylococcus epidermidis Ismarani, Diah; Pratiwi, Liza; Kusharyanti, Indri
Pharmaceutical Sciences and Research Vol. 1, No. 1
Publisher : UI Scholars Hub

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Abstract

Acnes vulgaris is a chronic inflammatory disease of sebaceous follicles caused by Propionibacterium acnes and Staphylococcus epidermidis. The antibacterial activity of Impatiens balsamina L. has been studied for many years. The aims of this study are to find out the antiacne effect from methanolic extract of stems and leaves of Impatiens balsamina L. that formulated in gel. Simplicia were extracted using soxhlet technique bu using methanol as solvent. Extracts were then formulated in gel in three variations of HPMC 4000 and Carbopol 934 as gel base with ratio of 70:30 (F1), 50:50 (F2) and 30:70 (F3). The determination of antiacne effect was done using disc diffusion method. Evaluation of physical and chemical properties of those gels includes organoleptic examination, spreadability testing, adhesion testing and pH testing. Data were analyzed using R version 2.14.1 package R-commander. Determination results showed the diameter of inhibition zone from FI, FII and FIII for P.acnes are 6.46 ±0.15 mm; 12.16 ± 0.35 mm; 16.13 ± 0.35 mm and for S. epidermidis sebesar 14.5 ± 0.47 mm; 16.56 ± 0.651 mm; 17.13 ± 0.44 mm. Analysis results showed that gel of FIII gave optimal antiacne effect, and significantly different from positive control (p < 0,05). The evaluation of gel showed that they have homogeny texture, good spreadability and adhesion, as well as the pH.