Kurkumin merupakan senyawa bioaktif utama dan pigmen kuning dalam rimpang kunyit dan temulawak (Curcuma sp.). Kurkumin memiliki aktivitas fisiologis dan farmakologis luas, salah satu di antaranya sebagai antifertilitas. Pentagamavunon-O (PGV-0) merupakan salah satu senyawa analog kurkumin yang telah mendapatkan hak paten sebagai anti-inflamasi dan dikenalkan dengan sebutan Molekul Nasional (MOLNAS). Penelitian ini bertujuan untuk mengkaji mekanisme kerja dan letak aksi kurkumin dan PGV-0 pada fungsi ovarium khususnya steroidogenesis, proliferasi, dan apoptosis pada kultur sel granulosa (KSG) berbagai ukuran folikel ovarium dengan rangsangan follicle stimulating hormone (FSH), luteinizing hormone (LH), dan/atau prostaglandin F_2α (PGF_2α). Penelitian eksperimental ini terdiri atas dua rancangan penelitian. Penelitian I, untuk mengkaji pengaruh kurkumin dan PGV-0 terhadap produksi progesteron (P) dan 17 β-estradiol (E₂), proliferasi, dan apoptosis pada KSG dengan rangsangan FSH, LH, dan/ atau PGF_2α. Penelitian II, untuk mengkaji letak kerja kurkumin atau PGV-0 pada penghambatan produksi P pada KSG setelah pemberian teofilin dengan rangsangan FSH dan/atau LH. Penelitian ini menggunakan sampel sel granulosa folikel ovarium babi yang dibedakan :menjadi folikel kecil (1 - 2 mm), sedang (> 2-5 mm), dan besar (> 5 mm). Preparasi kultur sel granulosa mengikuti metode Channing & Kammerman (1978) dengan menggunakan Dulbecco's modified Eagle's medium (DMEM) pada cawan kultur 48 sumuran dan chambers slide 8 kotakan. Setelah diinkubasi selama 72 jam (konfluen), diberikan perlakuan dan diinkubasi lagi selama 36 jam. Produksi P dan E₂ diukur dengan menggunakan metode enzymeimmunoassay (EIA) atau radioimmunoassay (RIA). Aktivitas proliferatif ditentukan dengan metode immunositokimia menggunakan antibodi-monoklonal proliferating cell nuclear antigen (MAb-PCNA) dan enzim avidin-biotin immunoperoxidase (ABIP). Apoptosis ditentukan dengan metode terminal deoxynucleotidyl transferase mediated dUTP-biotin hick end labelling (TUNEL). Data dianalisis menggunakan analisis varians (ANOVA), dan jika terdapat perbedaan rerata antarkelompok perlakuan dilanjutkan dengan komparasi multipel menggunakan Duncan's multiple range test (DMRT). Perbedaan dianggap bermakna jika nilai P < 0,05. Hasil penelitian ini menunjukkan bahwa: (1). Pemberian kurkumin mengurangi produksi P dan E₂ pada KSG folikel besar secara bermakna, tetapi pada KSG folikel kecil dan sedang berpengaruh tidak bermakna. Kurkumin mengurangi produksi P dan E₂ yang terangsang oleh FSH atau LH pada KSG berbagai ukuran folikel. Pemberian kurkumin setelah diberi PGF_2α, berpengaruh tidak bermakna terhadap produksi P dan E₂ oleh KSG berbagai ukuran folikel. Pemberian PGV-0 tidak berpengaruh terhadap produksi P dan E₂ pada KSG bebagai ukuran folikel. Pemberian PGV-0 menghambat produksi P dan E₂ yang terangsang oleh FSH atau LH pada KSG berbagai ukuran folikel. Pemberian PGV-0 pada KSG setelah mendapat PGF_2α tidak berpengaruh terhadap produksi P dan E₂ pada KSG berbagai ukuran folikel. (2). Kurkumin menghambat aktivitas proliferatif pada KSG berbagai ukuran folikel. Kurkumin menghambat aktivitas proliferatif yang terangsang FSH atau LH pada KSG berbagai ukuran folikel. Pemberian kurkumin setelah mendapat PGF_2α, tidak merangsang aktivitas proliferatif pada KSG berbagai ukuran folikel. Pemberian PGV-0 atau PGV-0 setelah mendapat FSH, LH, dan/atau PGF_2α tidak menghambat aktivitas proliferatif pada KSG berbagai ukuran folikel. Pemberian PGF_2α menghambat aktivitas proliferatif pada KSG folikel besar, tetapi pemberian PGF_2α berpengaruh tidak bermakna pada KSG folikel kecil dan sedang. (3). Kurkumin merangsang apoptosis pada KSG berbagai ukuran folikel. Kurkumin merangsang apoptosis pada KSG yang terhambat oleh FSH atau LH pada KSG berbagai ukuran folikel. Pemberian kurkumin pada KSG setelah mendapat PGF_2α, tidak merangsang apoptosis pada KSG berbagai ukuran folikel. Pemberian PGV-0 pada KSG atau pemberian PGV-0 setelah mendapat FSH, LH, dan/atau PGF_2α tidak menghambat apoptosis pada KSG berbagai ukuran folikel. (4). Kurkumin dan PGV-0 tidak mengganggu akumulasi cAMP setelah pemberian teofilin pada. KSG berbagai ukuran folikel ovarium. Berdasarkan hasil penelitian mIli dapat disimpulkan: (1). Pemberian kurkumin menghambat produksi P, E₂, proliferasi, dan meningkatkan apoptosis pada KSG folikel besar (mature). Pemberian PGV-0 berpengaruh tidak bermakna terhadap produksi P, E₂, proliferasi, dan apoptosis pada KSG berbagai ukuran folikel ovarium. (2). Pemberian kurkumin atau PGV-0 menghambat produksi P; E₂, proliferasi yang terangsang FSH atau LH, dan merangsang apoptosis yang terhambat FSH atau LH pada KSG berbagai ukuran folikel ovarium. (3). Pemberian kurkumin atau PGV-0 dengan PGF_2α tidak berpengaruh bermakna terhadap produksi P, E₂, proliferasi, dan apoptosis pada KSG berbagai ukuran folikel ovarium. (4). Letak kerja kurkumin dan PGV-0 pada hambatan produksi P terjadi setelah akumulasi cAMP.

Curcumin is the major bioactive component and yellow pigment isolated from the rhizome of perennial herb Curcuma sp. (turmeric). The pigment exhibits a wide range of physiological and pharmacological properties; one of these is an antifertility. Pentagamavunon-0 (PGVO) is one of the curcumin analogues and is known as MOLNAS (National Molecule). It has been patented as antiinflammatory drug, and is more potent and safe in compare to other curcumin analogues. The aims of this studies were examine the mechanism and site of action of curcumin and PGV-0 on steroidogenesis, proliferative activity, and apoptosis in cultured porcine granulosa cell from varying stage of follicular growth (in vitro) in the stimulation of follicle stimulating hormone (FSH), luteinizing hormone (LH), and/or prostaglandin F_2α (PGF_2α). This experimental research consists of two studies. The first study was designed to examine the effects of curcumin and PGV-0 to progesterone (P) and 17 β-estradiol (E₂) production, proliferative activity, and apoptosis in cultured porcine granulosa cells from varying stage of follicular growth in the stimulation of FSH; LH, and/or PGF_2α. The second study was designed to establish the site action of curcumin and PGV-0 on inhibitor of P production in cultured porcine granulosa cells from varying stage of follicular groWth in the' stimulation of theophylline with and without FSH and/or LH. Sample of granulosa cells were taken from porcine follicle ovaries and were,' harvested from follicular fluids and differentiated from small (1 - 2 mm), medium (> 2 - 5 mm), and large (> 5 mm) follicles. The cell were incubated in 48 wells plate and 8 chambers slide for 72 h, after treatment were added the wells returned were incubated for 36 h. Concentration of P and E₂ in the medium of culture were determined using enzymeimmunoassay (EIA) and radio-immunoassay (RIA) kit.- Proliferative activity was determined using monoclonal antibody of proliferating 'cell nuclear antigen (MAb-PCNA) and avidin-biotin immunoperoxidase enzyme (ABIP). Apoptosis was determined using terminal deoxy-nucleotidyl transferase mediated dUTP-biotin nick end labelling (TUNEL) method. Data was analysed by one-way ANOVA and when significant differences indicated the analysis is continued by multiple comparison using Duncan's multiple range test (DMRT). Differences between means at P < 0.05 were considered statistically significant. The results of the studies demonstrated that: (1). Curcumin decreases P and E₂ production by granulosa cells culture of large follicles. Curcumin decreases FSH or LH stimulates P and E₂ production by granulosa cells culture of all stages of follicular growth. Coincubation curcumin with PGF_2α, shows no significant effect on P and E₂ production in granulosa cells culture of all stages of follicular growth. Pentagamavunon-shows no significant effect on P and E₂ production by granulosa cells culture of all stages of follicular growth. Pentagamavunon-0 decreases FSH or LH stimulates P and E₂ production in granulosa cells culture of all stages of follicular growth. (2). Curcumin decreases proliferative activity on granulosa cells culture of large follicle. Curcumin decreases FSH or LH stimulates proliferative activity in granulosa cells culture of all stages of follicular growth. Pentagamavunon-0 or coincubation PGV-0 with FSH or LH shows no significant effect on proliferative activity in granulosa cells culture of all stages of follicular growth. (3). Curcumin increases apoptosis in granulosa cells culture of all stages of follicular growth. Curcumin decreased FSH or LH inhibits apoptosis in granulosa cells culture of all stages of follicular growth. Pentagamavunon-0 shows no significant effect on apoptosis in granulosa cells culture of all stages of follicular growth. (4). Curcumin and PGV-0 shows no significant effect on the accumulation of cAMP in granulosa cells culture of all stages of follicular growth. From these studies were concluded that: (1). Curcumin inhibits P and E₂ production and so the proliverative activity in mature follicle cultured granulosa cell, and stimulates apoptosis of granulosa cells regardless of follicular maturation stage. By contrast, PGV-0 shows no significant effect on P and E₂ production, proliverative activity, and apoptosis in cultured granulosa cells of all sizes follicle: (2). The increases P and E₂ production and proliferative activity in respons to the treatment with FSH or LH were reduced by the concomitant treatment with curcumin and PGV-0 in cultured granulosa cells of all sizes follicle. (3). Coincubation curcumin and PGV-0 with PGF_2α, shows no significant effect on P and E₂ production as well as the proliverative activity in granulosa cells culture of all stages of follicular growth. (4). Curcumin and PGV-0 act after the accumulation of cAMP in granulosa cells culture of all stages of follicular growth.

Kata Kunci : Kurkumin, Pentagamavunon-0, Anti-inflamasi, Curcuma sp.