THE ABILITY OF CURCUMIN AND 4-ARYL CURCUMIN DERIVATIVES AS RADICAL SCAVENGER OF SUPEROKSID

Superoxide radical scavenger property of curcumin and its derivatives was investigated using Nitro Blue Tetrazolium (NBT) method. Curcumin was modified at the active methyleen groups with aryl group (4-aryl curcumin), which supposed to be related to its anti-inflamatory activity. The experiment were carried out by mixing curcumin solution at five different concentrations and superoxide radical. After incubating for 60 minutes, NBT2+ (15,29 M) was added and measured at max of 687 nm, so as the curcumin derivatives. The result showed that superoxide radical made bathochromic shift of curcumin. Bathochromic shift was higher when aryl group was attached at active methyleen C-4 of curcumin. This shift made the max of curcumin and its derivatives moving close to max of diformasan (result of NBT2+ reduction by superoxide radical), producing an overlapping spectra. The increasing concentration of curcumin and its derivatives made the increasing max of diformasan absorbance. Thus, the method of NBT2+ reduction, in fact, unable to estimate the scavenger property of curcumin and its derivatives.Key words: curcumin derivatives, superoxide radica

Daya reduksi kurkumin dan turunannya (4-alkil-kurkumin) terhadap ion ferri yang diuji dengan metode orho-fenantrolin kompleks: The reduction ability of curcumin and its derivative (4-alkyl-curcumin) toward ferri ion and

Kurkumin yang mempunyai gugus atom C 4, diapit oleh gugus lcarbonil, menurut Tonnesemn dan Greenhill, mempunyai sifat reduktor. Penelitian ini untuk mengetahui kemampuan sifat reduktor kurkumin dan turunannya (4-alkil-kurkumin) terhadap ion ferri, yang diuji dengan orto-fenantrohn kompleks. Substitusi pada atom C-4, dengan gugus alkil mempunyai sifat sebagai pendorong elektron akan merubah dava reduksi kurkumin. Larutan kurkumin ditambah larutan dalam air atau metanol ferriklorida dan larutan ofenatrolin sebanyak 4.10-4M dicampur dan diencerkan sampai tertemtu. Percobaan lain dengan campuran yang sama ditambah larutan EDTA sebagai pengomplek untuk kontrol dan setelah 3 jam besarnya serapan dibaca pada 510 nm. Hasil menunjukkan bahwa kemampuan reduksi kurkunrin dan derivatnya mempunyai urutan sebagai berikut4-metil-kurkumin 4-benzil-kurkumin 4-isopropil-kurkumin -- kurkumin > 1,7-difenil,(1,6-heptadien 3,5-dion) atau kurkumin taktersubsitusi. Dart data yang diperoleh ternyata efek sterik gugus pensubstitusi atom C-4, lebih dominan pengaruhnya dart pada sifat pendorong elektron. Substitusi metil sifat pendorongnya paling lemah dan sifat strilarya terkecil, tetapi sifat reduksinya terbesar dibanding turunan kurkumin yang diuji. Hilangnya gugus metoksi dan gugus hidroksi pada cincin aromatis rnenurunkan daya reduksi terhadap ion ferri. Kata kunc: Turunan kurkumin, daya reduksi, gugus sterik alki

T47D cells arrested at G2M and Hyperploidy Formation Induced by a Curcumin’s Analogue PGV-1

its chemical structure than curcumin. As a curcumin analogue, PGV-1 was considered to have anticanceractivities. This research was conducted to study the effect of PGV-1 on the cycle progression of T47D cells. Cytotoxiceffects of PGV-1 on T47D cells were determined using MTT assay, and the the effect on cell cycle progressionwas carried out using flowcytometry. Western blot analysis was used to analyze protein expression correspondingto cell cycle progression. The result showed that at the concentration of 2.5 μM PGV-1 inhibited cell cycleprogression through G2/M arrest and induced of cells hyperploidy formation. The hyperploidy formation inducedby PGV-1 was related to the increase of cdc-2 expression. PGV-1 2.5 μM elevated the level of p21 CIP/KIPthrough p53- independent manner. Apoptosis was also induced by PGV-1 at early phase of treatment indicated byPARP cleavage due to activation of caspase-3/7 after 12 h treatment. The results above suggest that PGV-1 inhibitsthe growth of T47D cells targeted on microtubules.Keywords: PGV-1, G2/M arrest, apoptosis, p2

T47D cells arrested at G2M and Hyperploidy Formation Induced by a Curcumin’s Analogue PGV-1

its chemical structure than curcumin. As a curcumin analogue, PGV-1 was considered to have anticanceractivities. This research was conducted to study the effect of PGV-1 on the cycle progression of T47D cells. Cytotoxiceffects of PGV-1 on T47D cells were determined using MTT assay, and the the effect on cell cycle progressionwas carried out using flowcytometry. Western blot analysis was used to analyze protein expression correspondingto cell cycle progression. The result showed that at the concentration of 2.5 μM PGV-1 inhibited cell cycleprogression through G2/M arrest and induced of cells hyperploidy formation. The hyperploidy formation inducedby PGV-1 was related to the increase of cdc-2 expression. PGV-1 2.5 μM elevated the level of p21 CIP/KIPthrough p53- independent manner. Apoptosis was also induced by PGV-1 at early phase of treatment indicated byPARP cleavage due to activation of caspase-3/7 after 12 h treatment. The results above suggest that PGV-1 inhibitsthe growth of T47D cells targeted on microtubules.Keywords: PGV-1, G2/M arrest, apoptosis, p2

EFFECT OF ASPIRIN ON RAT LIVER CLASS- GLUTATHIONE S-TRANSFERASE ACTIVITY

Inflammation is the response of living tissues caused by injury. It involves a complex battery of enzyme activation, mediator release, extravasation of fluid, cell migration, tissue breakdown and repair. Inflammation mediator is synthesized in the body by several steps and catalyzed by several enzymes, such as cyclooxygenase and class- glutathione S-transferases (GSTs) in the cyclooxygenase arachidonic acid cascade. Aspirin, which has been reported as an inhibitor to inflammation mediator synthesis in the cyclooxygenase arachidonic acid cascade, is a non-steroid anti-inflammatory agent. The aim of this research is to study the effect of aspirin on rat liver class- GST activity in vitro. GSTs were isolated from the rat liver cytosolic fraction by centrifugation according to Lundgren. Protein concentration in the cytosol was determined spectrophotometrically using bovine serum albumin as a standard. The GST activity was determined using conjugation reaction rate between glutathione (GSH) and 1,2-dichloro-4-nitrobenzene (DCNB), followed by determining IC50 of aspirin. Then, a study was done to determine the ability of aspirin in inhibition of the rat liver class- GST activity in vitro. It can be concluded that aspirin has no inhibitory effect on rat liver class- GST activity.Key word: Aspirin, anti-inflammatory, glutathione S-transferase

Pengaruh Perlakuan PGV-1, PGV-0 dan Kurkumin terhadap Protein yang Terlibat dalam Siklus Sel Fase G2-M dan Apoptosis pada Sel Kanker Payudara T 47D

Previous experiments showed that curcumin analogue (PGV-1) inhibited breast cancer cell (T47D) growth at G2-M phase and induced cell apoptosis. This experiment was conducted to investigate the molecular effect of another curcumin analogue, PGV-0 and curcumin on the cell cycle progression and apoptosis as compared to the PGV-1. F lowcytometric method was conducted to analyze the effect of PGV-1 (2,5 µM), PGV-0 (5,0 µM) and curcumin (10,0 µM) on the cell distribution of various phase of T47D cell cycle. Western blot was also conducted to observe the effect of those compounds on proteins that involved in cell cycle (i.e. p21 and Cdc-Z) and apoptosis (Caspase-3/7/9). The results showed that PGV-1, PGV-0 and curcumin induced hyperploidy phenomenon on T47D cell. PGV-1 inhibited the cell cycle specifically on G2-M phase. Molecular observation showed that PGV-1 and PGV-0 were able to increase the expression of p21 protein and the Cdc-2 proteins, whilst curcumin was able to activate the Cdc-2 protein. The compounds have ability to induce apoptosis on T47D cell via Caspase3/7 activation. In conclusion, PGV-1, PGV-0 and curcumin inhibited the T47D cell cycle progression and induced cell apoptosis.Penelitian sebelumnya menunjukkan bahwa analog kurkumin PGV-1 dapat menghambat pertumbuhan sel kanker payudara pada fase G2-M dan menginduksi terjadinya apoptosis. Penelitian ini dilakukan untuk membandingkan pengaruh molekuler analog kurkumin PGV-0 dan kurkumin dengan PGV-l pada siklus sel dan apoptosis. Metode flowcytometry dilakukan dengan menganalisis pengaruh perlakuan PGV-1 (2,5 µM), PGV-0 (5,0 µM) dan kurkumin (10,0 µM) terhadap distribusi sel pada berbagai fase siklus sel T47D. Metode Western blot dilakukan untuk mengamati pengamh perlakuan senyawa uji terhadap protein yang terlibat pada siklus sel (p21 dan Cdc-2) dan apoptosis (Caspase-3/7/9). Hasil pengamatan pada perlakuan dengan PGV-1, PGV-0 dan kurkumin menunjukkan fenomena hiperploidi. Perlakuan dengan PGV-l secara spesifik menunjukkan fenomena penghambatan pada fase G2-M. Pengamatan molekuler menunjukkan PGV-1 dan PGV-0 mampu mempengaruhi ekspresi protein p21 dan Cdc-2 sedangkan kurkumin hanya mempengaruhi aktivasi Cdc-2. Ketiga senyawa uji terbukti mampu menginduksi apoptosis pada sel T47D melalui aktivasi Caspase-3/7. Secara keseluruhan, perlakuan ketiga senyawa uji mampu mempengaruhi siklus sel dan dapat menginduksi terjadinya apoptosis pada sel T47