Kurkumin merupakan pigmen kuning alami dari rimpang kunyit yang diduga memiliki aktivitas kemopreventif terhadap sel kanker melalui mekanisme jalur pensinyalan apoptosis. Penelitian ini bertujuan untuk menguji hubungan kurkumin terhadap kadar protein RASSF1A, Bax, dan aktivitas kaspase-3 dalam menunjang mekanisme apoptosis pada sel kanker payudara CSA03, MCF-7, dan MDA-MB-468.
Penelitian eksperimen in vitro dilakukan di laboratorium terpadu Fakultas Kedokteran Universitas Indonesia Jakarta, laboratorium terpadu Fakultas Kedokteran Universitas YARSI Jakarta, RSUPN Dr. Cipto Mangunkusumo Jakarta, serta RS Islam Jakarta tahun 2016–2018. Pemberian kurkumin terhadap sel kanker didasarkan atas perbedaan dosis dan waktu pemberian. Uji sitotoksisitas setelah pemberian kurkumin ditentukan secara MTS. Kadar protein RASSF1A dan Bax diuji secara ELISA. Aktivitas kaspase-3 digunakan untuk mengetahui apoptosis diuji secara flowsitometri. Selanjutnya perubahan morfologi sel diamati melalui pewarnaan acridine orange/ethidium bromide.
Pemberian kurkumin terhadap sel-sel yang diuji menunjukkan konsentrasi IC50 yaitu 40,85 µg/mL pada sel CSA03; 75,73 µg/mL pada sel MCF-7; dan 380,79 µg/mL pada sel MDA-MB-468. Pemberian kurkumin menunjang mekanisme apoptosis melalui jalur RASSF1A, Bax, dan aktivitas kaspase-3 pada sel kanker payudara.
Kata Kunci: Apoptosis, Bax, CSA03, kaspase-3, kurkumin, MCF-7, MDA-MB-468, pewarnaan ganda, RASSF1A
Curcumin is a natural yellow pigment from turmeric rhizome which is thought to have a chemopreventive effect on cancer through the mechanism of apoptotic signaling pathways. This study aims to examine the correlation of curcumin with protein level of RASSF1A, Bax, and caspase-3 activities in conjunction with the mechanism of apoptosis in CSA03, MCF-7, and MDA-MB-468 breast cancer cells.
In vitro experimental research was carried out at the Integrated Laboratory of Faculty of Medicine, Universitas Indonesia Jakarta; RSUPN. Dr. Cipto Mangunkusumo Jakarta; and Jakarta Islamic Hospital during 2016–2018. Curcumin was administered to the cancer cells in different doses and time. Cytotoxicity test after administration of curcumin was determined by MTS. The protein level of RASSF1A and Bax were measured by ELISA. Caspase-3 activity was used to determine apoptosis by flow cytometry. Furthermore, changes in cell morphology were observed by acridine orange/ethidium bromide staining.
The administration of curcumin to the cells showed IC50 concentrations of 40.85 µg/mL in CSA03 cells; 75.73 μg/mL in MCF-7 cells; and 380.79 µg/mL in MDA-MB-468 cells. The administration of curcumin supports the mechanism of apoptosis through the RASSF1A, Bax, and caspase-3 activity in breast cancer cells.
Indonesia adalah negara tropis dengan transmisi infeksi DENV yang tinggi dan sebuah ancaman kesehatan di dunia tanpa terapi spesifik yang dapat bekerja secara tunggal. Rakyat Indonesia memiliki kepercayaan tinggi atas obat herbal, salah satunya yang berasal dari Kunyit dengan senyawa utama, Kurkumin dengan efek antioksidan, pencegah kanker dan anti-inflamasi yang sudah terbukti melalui uji in vivo dan in vitro. Beberapa penelitian membuktikan bahwa kurkumin bekerja sebagai antivirus DENV-2 namun mekanisme yaitu waktu dimana kurkumin bekerja paling efektif, belum diketahui. Penelitian ini dilakukan secara in vitro dengan Sel Vero yang diinfeksikan DENV-2. Fokus pada penelitian ini adalah membandingkan mekanisme penghambatan replikasi DENV-2 sekaligus persentase viabilitas sel pada pre-post (whole) dan post infeksi setelah diberikan Kurkumin dengan dosis 20 ug/mL. Infektivitas hambatan dan viabilitas sel diteliti melalui metode focus assay dan MTT assay. Berdasarkan penelitian yang dilakukan, hasil penghambatan inefektivitas pada mekanisme pre-post (whole) dan post infeksi adalah 99,74% ± 3,90 dan 51,31% ± 8,97 secara berurutan. Penelitian untuk viabilitas sel mendapatkan hasil 73,21% dan 81,66% untuk pre-post (whole) dan post infeksi secara berurutan. Hasil penelitian menunjukan kurkumin memiliki efektivitas dalam mengambat DENV-2 lebih tinggi pada mekanisme pre-post infeksi (whole), dengan persentase penghambatan lebih tinggi serta toksitas rendah dengan viabilitas diatas 50%.
In Indonesia, DENV infection remains a global health threat without an effective therapy available. One of Indonesian’s herbal medicine, turmeric with Curcumin as its main compound is believed to have antioxidant, cancer-preventing and anti-inflammatory effects through in vivo and in vitro trials. Previous studies have shown that curcumin act as DENV-2 antivirus. However, its mechanism, namely the time at which curcumin work effectively, is not known. This research was conducted using DENV-2 infected Vero cells through in vitro method. The focus of this study was to compare the mechanism of DENV-2 replication inhibition as well as the viability of Cell in the pre-post (whole) and post-infection phases after administrating curcumin with a dose of 20 ug/mL. Focus assay and MTT assay methods were used in the experiment. Based on the research conducted, the results of ineffectiveness inhibition on the pre-post and post infection mechanisms were 99.74% ± 3.90 and 51.31% ± 8.97, respectively. The results for cell viability showed 73.21% and 81.66% for the pre-post (whole) and post-infection mechanisms, respectively. The results showed that curcumin is more effective in inhibiting DENV-2 in the pre-post infection mechanism (whole), with a higher percentage of inhibition and less toxicity with viability above 50%.
Pendahuluan.ÃÂ Kurkumin merupakan senyawa alami yang ditemukan pada akar tumbuhan Curcuma longa. Kurkumin memiliki sifat penyembuhan yang sangat baik, diantaranya termasuk anti-inflamasi, anti-bakteri, dan antioksidan. Telah dijelaskan pula pada beberapa studi bahwa kurkumin memiliki sifatÃÂ renoprotectiveÃÂ yang dapat membantu memperbaiki penyakit gagal ginjal kronik (CKD). Meskipun kurkumin memiliki banyak manfaat kesehatan untuk ginjal, jumlah kurkumin yang dapat mencapai jaringan ginjal sangatlah sedikit. Hal ini dikarenakan bioavailabilitas oral kurkumin hanya mencapai 1% yang disebabkan oleh buruknya absorpsi kurkumin pada saluran pencernaan. Penelitian ini bertujuan untuk meningkatkan konsentrasi kurkumin pada organ/jaringan ginjal dengan cara memperkecil ukuran partikel kurkumin menjadi nanocurcumin.
Metode.ÃÂ Penelitian ini menggunakan sediaan nanokurkumin yang dibuat dengan teknik ball milling. Dosis tunggal kurkumin atau nanokurkumin sebanyak 500 mg/kg diberikan secara oral kepada tikus Sprague-Dawley betina. Tikus didekapitasi pada menit ke-180 dan -240 setelah pemberian kurkumin atau nanokurkumin untuk pengambilan organ ginjal yang nantinya setiap 100 mg jaringan ginjal akan dihomogenisasi dengan larutan Normal Saline 0.9% sebanyak 1 ml. Homogenat jaringan ginjal akan dianalisa menggunakan UPLC-MS/MS dengan sumber ionisasi electrospray (ESI) positif.
Hasil.ÃÂ Konsentrasi kurkumin cenderung lebih tinggi dibandingkan konsentrasi nanokurkumin pada jaringan ginjal tikus setelah pemberian dosis tunggal kurkumin/nanokurkumin sebanyak 500 mg/kg pada jam ke-3 dan ke-4.
Kesimpulan.ÃÂ Kurkumin cenderung untuk memiliki konsentrasi yang lebih tinggi dibandingkan konsentrasi sediaan nanokurkumin pada jaringan ginjal tikus.
Introduction.ÃÂ CurcuminÃÂ is a naturally occurring compound found inÃÂ Curcuma longaÃÂ roots. It possesses great healing properties which mainly include anti-inflammatory, anti-bacterial, and anti-oxidative. It has also been described that curcumin has renoprotective effects and is proven to be able to ameliorate chronic kidney diseases (CKD). Despite having numerous health benefits for the kidney, the number of curcumin that can reach the kidney is very little, in respect to its low oral bioavailability which is only 1% due to poor absorption from the gastrointestinal tract. This study aims to enhance curcumin concentration in the kidney by decreasing curcumin particle size into nanocurcumin.ÃÂ
Methods.ÃÂ This study uses nanoparticle curcumin that is produced by using ball milling technique. A single dosage of 500 mg/kg curcumin or nanocurcumin was given orally to female Sprague-Dawley rats. Rats were decapitated at minute-180 and 240 after curcumin or nanocurcumin administration for kidney collection, which then homogenized with a ratio of 100 mg kidney tissue per 1 mL normal saline 0.9%. Kidney tissue homogenates were analyzed using UPLC-MS/MS with positive electrospray ionization (ESI).
Results.ÃÂ Curcumin concentration in rats kidney tissue tended to be slightly higher than nanoparticle curcumin after a single dose of 500 mg/kg curcumin or nanocurcumin at both 3 and 4 hours.
Conclusion.ÃÂ Curcumin has the propensity to have a higher concentration than nanocurcumin in rats kidney tissue.