Arleni Arleni
Department of Biology

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Influence of primaquine and ritonavir interaction on CYP3A4 mRNA expression in HepG2 cell culture Iskandarmudasyah, Adam; Louisa, Melva; Arleni, Arleni; Jusman, Sri W.A.; Suyatna, Franciscus D.
Medical Journal of Indonesia Vol 21, No 1 (2012): February
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (358.898 KB) | DOI: 10.13181/mji.v21i1.471

Abstract

Background: Concomitant treatment with antimalaria and antiretroviral drug is a new challenge in the management of malaria and HIV co-infection. Primaquine is a substrate and also an inhibitor of CYP3A4, while ritonavir is a substrate, an inhibitor, and also an inducer for CYP3A4. The objective of this study is to measure the CYP3A4 mRNA expression in HepG2 cell culture induced by primaquine and ritonavir co-treatment.Methods: For the initial study HepG2 cells were treated with 30, 40, 50 uM of primaquine; 2, 10, 20 uM ritonavir; DMSO ≤0.1 % for negative control; or 20 uM rifampicin for positive control. While for the co-treatment study the cells were treated with 40 uM primaquine+10 uM ritonavir; DMSO ≤0.1 %; or 20 uM rifampicin for 72 hours. The cells were harvested using trypsin–EDTA and total RNA was extracted using the Tripure isolation reagent. After determining the quantity of RNA spectrophotometrically, CYP3A4 mRNA expression was quantified using real-time reverse transcription polymerase chain reaction (RT-PCR).Results: The expression of CYP3A4 mRNA was up-regulated (1.22 fold over control) in HepG2 cells co-treated with primaquine and ritonavir. These data suggest that the induction effect of ritonavir was more dominant than the inhibitory effect of primaquine.Conclusion: Concomitant administration of primaquine and ritonavir result in up-regulation of CYP3A4 mRNA expression in vitro. (Med J Indones 2012;21:3-7)Keywords: CYP450 induction, CYP3A4, drug interaction, primaquine, ritonavir
Cytology technique: development of a simple spot method for cultured cell suspension Pawitan, Jeanne A.; Damayanti, Lia; Arleni, Arleni; Swantari, Ni M.
Medical Journal of Indonesia Vol 19, No 1 (2010): February
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (722.566 KB) | DOI: 10.13181/mji.v19i1.378

Abstract

Aim To develop a simple spot method to attach cultured cells in suspension on to a glass slide.Methods We compared three approaches using both conventional and special glass slide (Shandon-Polysin), either without additional fetal bovine serum (FBS), or with addition of 3 or 10 μl of FBS to a 20 μl sample (altogether there were six approaches). The slides were examined qualitatively for the background color, boundary color and intactness, and whether there were folded and detached parts. Further, for each slide, the attached intact cells were counted, and the percentage of attached intact cells per number of spotted cells was calculated. The difference in attach intact cells between different approaches was analyzed by ANOVA using SPSS 13.0 for windows.Results There were no significant difference in the percentage of attached intact cells between the six approaches (P= 0.804), though the approach using special glass slide without additional FBS (FBS final concentration 5%) yield the highest percentage of attached intact cells, showed clean background without folded parts.Conclusions We have developed a simple spot method for cultured cell suspension, and the best approach to make spot specimen is using special glass slide with 5% FBS in the cell suspension. (Med J Indones 2010; 19:26-31)Keywords: spot specimen, special glass slide, fetal bovine serum
Efek zat aromatase inhibitor dan GnRH agonis terhadap kadar Vascular Endothelial Growth Factor-A pada kultur jaringan endometriosis AS’ADI, A.S.; HESTIANTORO, A.; ARLENI, ARLENI
Indonesian Journal of Obstetrics and Gynecology Volume. 32, No. 1, January 2008
Publisher : Indonesian Socety of Obstetrics and Gynecology

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Abstract

Tujuan: Menganalisa efek zat aromatase inhibitor, GnRH agonis dan kombinasi keduanya terhadap kadar Vascular Endothelial Growth Factor-A (VEGF-A) pada kultur jaringan endometriosis dalam lingkungan kadar steroid seks yang berbeda. Tempat: RS Fatmawati, RSUPN Dr. Cipto Mangunkusumo, Klinik Kesehatan Reproduksi Raden Saleh Jakarta dan Laboratorium MAKMAL FKUI. Rancangan/rumusan data: Penelitian eksperimental. Bahan dan cara kerja: Selama kurun waktu Juni 2006 - April 2007, terkumpul 15 sampel jaringan endometriosis dari 15 pasien endometriosis. Sampel yang didapat berasal dari dinding kista endometriosis dan dari bercak-bercak endometriosis pada genitalia interna. Semua sampel diolah sesuai protokol yang dibuat. Ada 4 sampel (27%) yang berhasil tumbuh baik dalam medium kultur. Dari 4 sampel tersebut hanya 3 sampel yang mendapat perlakuan. Masing-masing sampel dibagi ke dalam 7 well dengan jumlah sel pada masing-masing well 7,1 - 9,1 x 103 sel/ml : well 1 ditambahkan Testosteron 100 nM/L, well 2 ditambahkan Testosteron 100 nM/L dan Estradiol 10 nM/L, well 3 ditambahkan Testosteron 100 nM/L, Estradiol 10 nM/L dan Letrozol (aromatase inhibitor) 10 nM/L, well 4 ditambahkan Testosteron 100 nM/L dan Letrozol 10 nM/L, well 5 ditambahkan Testosteron 100 nM/L, Letrozol 10 nM/L dan Leuprolide asetat (GnRH agonis) 100 ng/ml, well 6 ditambahkan Testosteron 100 nM/L dan Leuprolide asetat 100 ng/ml, well 7 tanpa perlakuan (kontrol). Setelah diinkubasi selama 72 jam, supernatannya diambil dan dilakukan pemeriksaan kadar VEGF-A dengan teknik ELISA. Hasil: Nilai median kadar VEGF-A yang paling tinggi terjadi pada pemberian Testosteron + Estradiol yaitu 20,228 pg/ml dan ini lebih tinggi bila dibandingkan kontrol yang hanya 9,233 pg/ml, maupun dengan sampel yang hanya diberikan Testosteron saja yaitu 9,944 pg/ml. Nilai median kadar VEGF-A pada sediaan yang diberikan Testosteron + Estradiol yaitu 20,228 pg/ml, bila dibandingkan dengan sampel yang mendapatkan perlakuan yang sama dan ditambahkan Letrozol (aromatase inhibitor) terjadi penurunan menjadi 14,205 pg/ml. Nilai median kadar VEGF-A pada sampel yang diberikan Testosteron + Letrozol (aromatase inhibitor) 16,335 pg/ml, Testosteron + Letrozol + Leuprolide asetat (GnRH agonis) 10,653 pg/ml dan Testosteron + Leuprolide asetat 11,364 pg/ml. Nilai terendah terjadi pada sampel yang diberikan Aromatase inhibitor + GnRH agonis. Kesimpulan: Nilai median kadar VEGF-A cenderung meningkat pada sampel yang diberikan Testosteron dan Estradiol. Nilai median kadar VEGF-A cenderung lebih rendah pada sampel yang diberikan kombinasi aromatase inhibitor dan GnRH agonis. [Maj Obstet Ginekol Indones 2008; 32-1: 11-21] Kata kunci: kultur jaringan endometriosis, testosteron, estradiol, letrozol, leuprolide asetat, VEGF-A, ELISA