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Action Target of Curcumin and PGV-0 on Luteal Cells Steroidogenesis through the Expression of Cytochrome P450scc

Journal of the Indonesian Medical Association Vol. 62 No. 4 April 2012
Publisher : Journal of the Indonesian Medical Association

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Abstract

Introduction: Curcumin and its analog pentagamavuon-0/PGV-0 can inhibit steroidogenesis of luteal cell culture. However, the action site of the curcumin and PGV-0 in luteal cells steroidogenesis is still unknown.Objectives: To analyze the action site of curcumin and PGV-0 in luteal cells steroidogenesis through its influences on the expression of cytochrome P450scc enzyme.Methods: PMSG-induced corpus luteums of Sprague Dawley rat were used as the subjects. Curcumin and PGV-0 with a dosage of 100 µM was given shortly after the stimulation of LH and/or PGF2a. Immunohistochemistry staining was done to assess the expression of cytochrome P450scc.Results: LH stimulation (50 ng/ml) increased the expression of cytochrome P450scc significantly (p<0.05) compared with the control group. In the other hand, PGF2a (0.56 µM) inhibited the expression of cytochrome P450scc significantly. Curcumin (100 µM) and PGV-0 (100 µM) inhibited the expression of cytochrome P450scc in the control group and groups stimulated by LH and/or PGF2a.Conclusion: Curcumin and PGV-0 inhibit the upstream transduction signal of cytochrome P450scc in the steroidogenesis of luteal cell culture. J Indon Med Assoc. 2012;62:136-41.Keywords: P450scc, steroidogenesis, transduction signal

Aktivitas Sitotoksik Ekstrak Etanolik Rimpang Temu Kunci (Boesenbergia pandurata) Terhadap Sel Kanker Serviks HeLa dan Sel Kanker Kolon WiDr

Majalah Kesehatan Pharmamedika Vol 3 N0 1 2011
Publisher : Majalah Kesehatan Pharmamedika

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Abstract

Temu kunci (Boesenbergia pandurata) rhizome showed cytotoxic effect against T47D breast cancer cell line. It contains Panduratin, a chalcone compund, that has been investigated as chemopreventive agent. The exploration of extract of temu kuci as chemopreventive agent was expected to be an alternative for cancer therapy. The aim of this research was to determine the cytotoxic activities of ethanolic extract of temu kunci against HeLa cervix cancer and WiDr colon cancer cell line. The cytotoxic activities of ethanolic extract of temu kunci were tested using MTT assay against HeLa and WiDr cells. The IC50 values were obtained using linier regression equation. The ethanolic extract of temu kunci showed cytotoxic activities on HeLa cervix cancer and WiDr colon cancer cell lines with IC50 at 87 µg/mL and 76 µg/mL, respectively. Low IC50 values (<100 µg/mL) showed that ethanolic extract of temu kunci is potential to be developed as chemoprevention agent on cervix cancer and colon cancer. However, its molecular mecahanism need to be explored.

Aktivitas Antiproliferasi Ekstrak Etanolik Herba Ciplukan (Physalis angulata L.) Terhadap Sel Hepar Tikus Betina Galur Sprague Dawley Terinduksi 7,12-Dimetilbenz[a]antrasena

Majalah Kesehatan Pharmamedika Vol 3 N0 1 2011
Publisher : Majalah Kesehatan Pharmamedika

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One of the natural materials as potentially efficacious chemopreventive agents are Ciplukan (Physalis angulata L.). Several previous studies reported that Physalis angulata L. herbs ethanolic extract (PEE) has cytotoxic activity and induction of apoptosis in breast cancer cells MCF-7 and HeLa cervical cancer cells. This study aims to determine the effects of  PEE as an chemopreventive agent on rat liver cells induced 7,12-dimetilbenz[a]anthracene (DMBA). This study used Sprague Dawley strain female rats aged 40-50 days were divided into 5 groups : (1) DMBA control group, mice were induced with DMBA in per oral dose of 20 mg/kg; (2) DMBA + PEE dose 750 mg/kgBW group ; (3) DMBA + PEE dose 1500 mg/kgBW group ; (4) solvent control group of CMC-Na 0,5%; (5) PEE dose 1500 mg/kgBW control group. PEE was dissolved in CMC-Na 0,5% and administered daily, starting the seventh week after administration of DMBA. At the beginning of  the tenth week of the study, rats were necropted and liver organs were isolated and stored in buffered formalin 4%. Qualitative analysis to determine the histopathology of liver cells through staining method of Hematoxyllin & Eosin (HE), while quantitative analysis to determine the level of liver cell proliferation by AgNOR staining method. The results showed in the DMBA control group that liver cell morphology changes that hiperproliferation leading to carcinogenesis. In DMBA + PEE dose of  1500 mg/kgBW group improved the situation of DMBA-induced liver cells histopathology and antiproliferation activity better than DMBA + PEE dose of 750 mg/kgBW on DMBA-induced rat liver cells. The results showed that Physalis angulata L. herbs ethanolic extract can inhibit cell proliferation in rat liver caused by DMBA administration through antiproliferation mechanism and have potential for the development as chemoprevention material on liver cancer

The Action Target of Pentagamavunon-0 on the Luteal Cell Steroidogenesis Through Phosphorylation Measurement of Extracellular Signal Regulated Kinase

Journal of the Indonesian Medical Association Vol. 63 No. 2 February 2013
Publisher : Journal of the Indonesian Medical Association

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Abstract

Introduction: Pentagamavunon-0 (PGV-0) compound, a curcumin analog, can inhibit steroidogenesis of luteal cell culture by inhibiting the production of progesterone. The action site of PGV- 0 on steroidogenesis of luteal cells is still unknown. The objective of this study is to analyze the effect of PGV-0 on the Extracellular Signal Regulated Kinase (ERK) phosphorylation in steroidogenesis of luteal cell culture.Methods: This study used corpus luteum of rat Sprague Dawley strain that was induced withPregnant Mares Serum Gonadotropine (PMSG). Pentagamavunon-0 was given shortly after the stimulation of LH and or PGF2a with or without forskoline. The phosphorylation of ERK was assessed by IHC method.Results: The result showed that LH stimulation increased ERK phosphorylation significantly.However, PGF2a decreased ERK phosphorylation significantly. Forskoline increased ERKphosphotylation significantly but no significant effect was found with LH stimulation. PGV-0inhibited ERK by either LH or forskoline stimulation.Conclusion: Pentagamavunon-0/PGV-0 can inhibits signal transduction of steroidogenesis lutealcells by acting on the up stream to ERK. J Indon Med Assoc. 2013;63:52-7Keywords: luteal cell, ERK, immunohistochemisty, forskolin

ARECA (Areca catechu L.) SEEDS ETHANOLIC EXTRACT AND ITS CHLOROFORM FRACTION INCREASE APOPTOTIC EFFECT OF DOXORUBICIN ON HUMAN COLON CANCER CELLS

Jurnal Bahan Alam Indonesia Vol 6, No 5 (2008)
Publisher : Indonesian Research Gateway

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Abstract

Pada penelitian terdahulu, ekstrak etanolik biji buah pinang (AE) dan fraksi kloroformnya (ACF) memiliki aktifitas sitotoksik pada sel kanker kolon WiDr dengan nilai IC50 masing-masing sebesar 124 μg/mL dan 119 μg/mL. Penelitian ini bertujuan untuk mengetahui efek kombinasi AE dan ACF dengan agen kemoterapi Doxorubicin (Dox), serta efek kombinasi tersebut dalam menginduksi apoptosis pada sel kanker WiDr.Serbuk biji buah pinang diekstraksi menggunakan etanol 96%. Kemudian ekstrak difraksinasi secara partisi menggunakan heksan dan klorofon untuk mendapatkan fraksi klorofom. Aktivitas sitotoksik baik AE, ACF dan Dox tunggal maupun kombinasi dilakukan dengan metode MTT. Pengamatan apoptosis menggunakan metode double staining (acrydine orange-ethidium bromide). Metode imunositokimia digunakan untuk mendeteksi ekspresi Bcl-2 dan COX-2.Kombinasi AE dan ACF dengan Dox menunjukkan efek sinergistik pada sel kanker WiDr dengan indeks kombinasi (Cl) <0.9 (P<0.05). Pada kombinasi Dox - AE, efek sinergis paling kuat terlihat pada kombinasi Dox 100 dan 334 nM dengan AE 60 μg/mL (Cl<0.1), sedangkan efek sinergis paling kuat pada kombinasi Dox-ACF terlihat pada kombinasi Dox 100 nM dengan ACF 30,40 dan 60 μg/mL (Cl=0.1). Kedua kombinasi tersebut mampu  menginduksi apoptosis pada sel kanker WiDr. Kombinasi Dox-ACF mampu menurunkan ekspresi Bcl-2 dan meningkatkan ekspresi Bax. Hasil tersebut menunjukkan apoptosis pada sel kanker WiDr yang dimungkinkan mekanismenya berhubungan dengan ekspresi Bcl-2 dan COX-2. 

PENINGKATAN EFEK SITOTOKSIK DOXORUBICIN OLEH NARINGENIN MELALUI PEMACUAN APOPTOSIS SEL KANKER PAYUDARA MCF-7

Jurnal Bahan Alam Indonesia Vol 7, No 3 (2010)
Publisher : Indonesian Research Gateway

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Abstract

Chemoterapic agent doxorubicin has a lot of side effects and resistance problem. Therefore, it is important to develop combination treatment of doxorubicin with natural product (co-chemotheraphy) such as naringenin. Naringenin has been proven to have cytotoxic activity against several cancel cell lines. Naringen also increase the sensitivity of MCF-7 breast cancer cell towards doxorubicin. The aim of this research is to examine the synergistic effect of naringenin on the cytotoxic activity of doxorubicin through apoptosis induction of MCF-7 cancer cell lines.The cytotoxic assay of naringenin and doxorubicin were carried out by MTT method using naringenin concentrations of 100-1250 μM and doxorubicin concentrations of 50-800 nM to determine the IC50. Based on the IC50 values, the cytotoxic assay of their combination were carried out  by MTT method using concentration of ⅛, ¼, ⅜ and ½ IC50 values. Apoptosis assay of their best combination concentrations were done using double staining method.Naringenin and doxorubicin showed cytotoxic effect on MCF-7 breast cancer cell with IC50 of 520 μM and 467 nM, respectively. Based on CI values, almost all concentration combination of naringenin and doxorubicin showed synergistic effect (CI 0,1-0,9). This synergistic effect is due to the induction of apoptosis, showed by the increasing number of orange fluorescence cells in double staining method. Based on this results, naringenin was potential to be delevoped as co-chemotherapeutic agent. However, the molecular mechanism need to be explored further.

INDUKSI APOPTOSIS EKSTRAK ETANOL CIPLUKAN (Physalis angulata L.) PADA SEL KANKER LEHER RAHIM HeLa MELALUI PENEKANAN EKSPRESI Bel-2

Jurnal Bahan Alam Indonesia Vol 7, No 7 (2011)
Publisher : Indonesian Research Gateway

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Abstract

The incidence of cervical cancer continue to rise encourage a variety of studies to explore the plant as chemopreventive. Ciplukan (Physalis angulata L.) is one of potential plant as chemopreventive agent by regulating cell proliferation, cell cycle and apoptosis. In this study the potential effect of ciplukan ethanolic extract as inducers of apoptosis in human cervical carcinoma HeLa cell line wil be futher investigated. Induction of apoptosis was evaluated by Double Staining methods and Immunocytochemistry. Apoptosis observations qualitatively indicate the occurrence of early apoptosis marked by orange fluorescence and apoptosis bodies because of cell fragmentation. Then the expression of Bcl-2 was observed to know molecular mechanism of apoptosis. Immunocytochemistry experiment showed that Bcl-2 as antiapoptosis protein was inhibited in comparison with control cells. These data indicated that ciplukan ethanolic extract (P.angulata L.) could induced the apoptosis Hela cell line by downregulation of Bcl-2. Study of ciplukan shoud be developed to support ciplukan to be effective anticancer agents.

Structure Modification of Ethyl p-methoxycinnamate Isolated from Kaempferia galanga Linn. and Citotoxicity Assay of The Products on WiDr Cells

Indonesian Journal of Cancer Chemoprevention Vol 1, No 1 (2010)
Publisher : Indonesian Research Gateway

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Abstract

Ethyl p-methoxycinnamate, major ingredient of Kaempferia galanga rhizome, have been reported not only has analgesic – anti inflammatory activities like NSAIDs which inhibited cyclooxygenase, but also inhibit tumor cell proliferation in specimen of mouse epidermis. Therefore, it will be interesting to carry out  synthetic studies on the derivates of ethyl  p-methoxycinnamate and searching their citotoxic activity on WiDr cell. We wish to report of structure modification on carboxyl moiety of  ethyl p-methoxycinnamate  and  evaluation on their citotoxic activity  on WiDr cell. Isolation of ethyl p-methoxycinnamate from Kaempferia galanga rhizome was carried out by percolation with ethanol 96% as solvent. Hydrolysis of ethyl p-methoxycinnamate in basic condition was performed to obtain p-methoxycinnamic acid. Preparation of some thiourea derivates of ethyl  p-methoxycinnamate was carried out  by microwave irradiation. Citotoxicity assay was carried out by MTT method for 48 h.   Modification  of  carboxyl  group  of  ethyl  p-methoxycinnamate to its thiourea form could be carried out by microwave irradiation gave; (E)-3-(4-methoxyphenyl)-N-(phenylcarba- mothioyl)acrylamide (50%); (E)-3-(4-methoxyphenyl)-N-(4-methoxyphenylcarbamothi- oyl)acrylamide (26%) and (E)-3-(4-methoxyphenyl)-N-(4-methylphenylcarbamothioyl) acrylamide (54%), yield calculated for 2 step from the acid chloride. All compounds showed no citotoxic effect on WiDr cell at 48 h incubation.

SynergisticCombinationofCiplukan(Physalis angulata) HerbsEthanolicExtractandDoxorubicinonT47DBreast CancerCells

Indonesian Journal of Cancer Chemoprevention Vol 1, No 1 (2010)
Publisher : Indonesian Research Gateway

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Abstract

Doxorubicinisoneofchemotherapeuticagentwidelyusedinbreastcancertreatment,but in high dose doxorubicin gives negative side effect, including vomit, nausea, immune suppression, and cardiac toxicity. This toxicity hopefully could be reduced by combination chemotherapy using natural herbs such as ciplukan herb. This research was conducted to explorecytotoxicactivityofsingleciplukanherbsethanolicextractanditscombinationwith doxorubicinonT47Dbreastcancercells.Cytotoxicactivityofciplukanherbsethanolicextract only and its combination with doxorubicinwere tested on T47D cells using MTT assay toobtainIC50valueandcombinationindex(CI),respectively.Singleextractshowedcytotoxic activityonT47DcellswithIC50valueofwas160*g/ml.Thus,combinationtreatmentfrom ciplukanherbsethanolicextractanddoxorubicinshowedsynergisticeffect(CI<1,0).Thiseffect wasreachedatconcentrationofciplukanherbsethanolicextract-doxorubicin80μg/ml-2nM, 80 μg/ml-4 nM, and 80 μg/ml-8 nM. This research indicated that ciplukan herbs ethanolic extractispotentialtobeappliedasco-chemotherapeuticagentinbreastcancertherapy.

Leunca (Solanum nigrum L.) Herbs Ethanolic Extract Increase Cytotoxic Activity of Cisplatin on Hela Cervical Cancer Cells

Indonesian Journal of Cancer Chemoprevention Vol 1, No 1 (2010)
Publisher : Indonesian Research Gateway

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Abstract

Cervical cancer is one of leading causes of cancer death in women in the developing countries. The use of cisplatin as chemotherapy agent in cervical cancer is known to cause side effects and also resistance for long-term uses. One of the strategies to prevent cervical cancer based on combination agents is being developed. Leunca (Solanum nigrum L.) has been revealed to inhibit growth of human cancer cells. Therefore, it can be used in combination with cisplatin to reduce those side effects and prevent the occurrence of cell resistance. Ethanolic extract of Leunca Herb (ELH) and cisplatin were tested their cytotoxic effect on HeLa cervical cancer cell by using MTT assay to determine IC50 value. The combinationss of cisplatin-ELH were tested to determine the combination index (CI value).  The IC50 of ELH and cisplatin on HeLa cells were 227 µg/mL and 17 µM. rRespectively. Tthe study of combination resulted that almost all the index combinations were <0,9 showed  the effect of synergism combination. The Ooptimum concentration of combination was  1/8 IC50 cisplatin–1/8 IC50 ELH. The results indicated that ELH had a potency to be combination agent to enhance the activity of cisplatin on HeLa cervical cancer cells. Therefore, further study on its molecular mechanism needs to be explored.

Co-Authors . Anindyajati . Larasati . Sarmoko . Sugiyanto . Sukardiman Adam Hermawan Aditya Fitriasari Agung Endro Nugroho Ainun Wulandari, Ainun Ameilinda Monikawati Andita Pra Darma Arief Nurrochmad Astrid Ayu Maruti Aulia Katarina B. Sudarto Barinta Widaryanti Broto Kardono Chandra Risdian Dewi Arum Sekti, Dewi Arum Dewi Pratiwi Djaswadi Dasuki Dwi Ana Nawangsari, Dwi Ana Dwi Nurahmanto, Dwi Dyaningtyas D. P. Putri Dyaningtyas Dewi Pamungkas P, Dyaningtyas Dewi Dyaningtyas Dewi Pamungkas Putri Dyaningtyas Dewi Putri, Dyaningtyas Dewi Endah Puji Septisetyani, Endah Puji Endah Puspitasari Endang Purwaningsih Erlina Rivanti Erna Prawita Setyowati Fany Mutia Cahyani Fikri Amalia Fina Aryani Goenadi, Fina Aryani Fitria Rahmi Fortunella Tjondro Fransiscus Feby Handoko HENDRI WASITO Herwandhani Putri Ibrahim Arifin Ika Nurzijah Ika Rahmawati Sutejo Ilham Agusta Fauzi Indri Kusharyanti Inna Armandani, Inna Inna Armandari Iwan Sahrial Hamid JAKA WIDADA Juni Ekowati Kartika Dyah Palupi Kholid Alfan Nur Lany Candra, Lany Laras Widawaty Putri Luthfia Indriyani Marcellino Rudyanto Maria Dwi Supriyati, Maria Dwi Marissa Angelina Masashi Kawaichi, Masashi Maya Fitria Moordiani ., Moordiani Muhammad Da’i, Muhammad Muhammad Fithrul Mubarok, Muhammad Fithrul Muthi Ikawati Muthi’ Ikawati Nanda Resa Pratama Niken Nur W, Niken Novi Hastuti, Novi Nunuk Aries Nurulita Nurma Sabila Perdana Adhi Nugroho R A Susidarti Rachmaniar Rahmat, Rachmaniar Raditya Prima Istiaji Rahmi Khamsita Ratih Hardika Pratama Ratna Asmah Susidarti Retno Murwanti Retno Murwantib Rina Andriyani Riris I Jenie, Riris I Riris Istighfari Jenie Rita Riata Rosa Adelina Rosana Anna Ashari, Rosana Anna Rosita Melannisa, Rosita Sari Haryanti Sendy Junedi, Sendy Sendy Junedy, Sendy Shigeru Sasaki Shirly Kumala Sismindari, ., Sismindari, Sitarina Widyarini Sofa Farida Sri Handayani Sri Kadarsih Soejono Sri Kasianningsih Sri Susilowati Sri Tasminatun, Sri Sudarsono . Sugiyanto . Supardjan A. M., Supardjan A. Supardjan AM, Supardjan Susi Ari Kristina Suven ., Suven Tutuk Budiati Umar A. Jenie Umar Anggara Jenie Yurista Gilang Yurista Gilang Ikhtiarsyah Yuyun Farida, Yuyun Zalinar Udin