| dc.description.abstract | Penyakit degeneratif adalah penyakit yang risikonya meningkat seiring dengan meningkatnya usia. Salah satu penyebab penyakit degeneratif adalah timbulnya radikal hidroksil dalam mekanisme biokimia di dalam tubuh.Kanker payudara merupakan salah satu penyakit degeneratif penyebab kematian terbesar di dunia pada wanita.Resistensi agen kemoterapi menyebabkan perlu dilakukannya pencarian bahan alam dengan aktivitas antikanker. Penggunaan bahan alam diharapkan dapat meningkatkan efektivitas dan menurunkan efek samping.Tujuan penelitian ini untuk mengetahui aktivitas antikanker dan antioksidan fraksi etilasetat buah andaliman (FEABA) dan herba poguntano(FEAHP) secara in vitro dan in vivo.
Fraksi diperoleh melalui maserasi bertingkat dengan menggunakan pelarut n-heksana dan etilasetat. Sel T47D dibiakkan dalam media kultur RPMI , dan sel Vero dibiakkan dalam media kultur M199 kemudian diberi FEABA atau FEAHP dan diberikan larutan H2O2 0,8 mM. Pengujian sitotoksik secara in vitro menggunakan metode MTT [3-(4,5-dimetiltiazol-2-il)-2,5 difeniltetrazolium bromida] yang kemudian dianalisis menggunakan SPSS 22. . Pengujian apoptosis, siklus sel, ekspresi kaspase 3/7, ROS, permeabilitas membran mitokondria, ekspresi protein PI3K, mTOR, siklin D1 dan p53 dengan metode flow sitometri. Pengujian ekspresi Bcl-2 dengan metode imunositokimia, analisis fragmentasi DNA dengan metode elektroforesis, analisis morfologi inti dengan metode pewarnaan inti dan analisis ekspresi gen PI3KCA, Akt-1, Akt-2, mTOR, EGFR, VEGFR-2, ERα dan p53 dengan metode RT-PCR. Pengujian aktivitas antioksidan secara in vitro dilakukan dengan metode DPPH, ABTS dan reducing power serta secara in vivo dengan metode SOD pada tikus putih betina yang diinduksi dengan doksorubisin. Penentuan kandungan fenol total dengan metode Folin-Ciocalteu dan kandungan flavonoid total dengan metode kolorimetri.
Hasil uji sitotoksik (IC50) FEABA dan FEAHP sebesar 48,94 ± 0,32μg/mL dan 62,53 ± 0,55μg/mL. Fraksi etilasetat herba poguntano dan buah andaliman memiliki aktivitas antikanker melalui penghambatan siklus sel dengan akumulasi pada fase G0-G1 (57,23% dan 60,48%), pemacuan apoptosis awal (72,75% dan 70,12%) dan apoptosis akhir (10,23% dan 8,54%), ekspresi kaspase 3/7 (35,38% dan 80,41%), ekspresi ROS (76,78% dan 76,81%), permeabilitas membran (11,68% dan 19,02%), menurunkan ekspresi protein Bcl-2 dan siklin D1 (45,94% dan 69,19%), meningkatkan ekspresi p53 (8,10% dan 12,83%), menurunkan ekspresi protein PI3K (2,93% dan 4,71%) dan mTOR (14,03% dan 42,83%), meningkatkan aktivitas fragmentasi DNA, menyebabkan perubahan morfologi sel T47D dan menurunkan ekspresi gen PI3KCA, Akt-1, Akt-2, mTOR, EGFR, VEGFR-2, ER-α dan meningkatkan ekspresi gen p53.Viabilitas sel pada uji sitoprotektif FEAHP dan FEABA pada konsentrasi 100 μg/mL adalah88,83 ± 2,90% dan 87,03 ± 1,36%. FEAHP dan FEABA memiliki aktivitas sitoprotektif menurunkan ekspresi ROS (62,58% menjadi 54,63% dan 58,22%). Aktivitas antioksidan dengan metode DPPH nilai IC50 (166,90 ± 0,10 dan 156,48 ± 0,04 μg/mL), persen peredaman metode ABTS pada konsentrasi 200 μg/mL (35,21 ± 0,03% dan 45,78 ± 0,21%), absorbansi pada metode reducing power (0,2172 ± 0,0002 dan 0,4044 ± 0,0002). Kandungan fenol dan flavonoid total FEAHP dan FEABA (92,88 ± 0,50mg/g dan 90,41 ± 0,06mg/g) dan (84,39 ± 0,07mg/g dan 77,56 ± 0,02mg/g). Kadar SOD pada tikus putih betina pemberian dosis 300 mg/Kg BB (3,242 ± 0,086 dan 5,644 ± 0,152 U/mL). Pada pengamatan histopatologi jaringan hati terlihat pengurangan kerusakan jika dibandingkan dengan kontrol. Penelitian ini menunjukkan bahwa FEABA dan FEAHP berpotensi sebagai antikanker dan antioksidan dimana FEABA lebih berpotensi jika dibandingkan dengan FEAHP. | en_US |
| dc.description.abstract | Degenerative disease is used as a medical term to describe a process of decline in cell function without a known cause, from a previously normal to worse. One of the causes of degenerative diseases is the appearence of hydroxyl radicals in the biochemical mechanisms in the body. Breast cancer is one of the world's leading cause of death in women. Due to the resistance of chemotherapeutic agents,there is a continuous need to search of natural products withanticancer activity. The use of natural products is expected to increase the effectiveness and decrease side effect. The purpose of this study was to investigate the anticancer and antioxidant activities of ethylacetate fraction of andaliman fruits (FEAAF) and poguntano herbs (FEAPH)in vitro and in vivo.
The fractions were obtained by maceration using n-hexane and ethylacetate solvents. T47D cells were grown in culture medium RPMI then given by FEAPH and FEAAF. Vero cells were cultured in M199 culture medium then FEAPH and FEAAF and given 0.8 mM H2O2 solution. In vitro cytotoxic activity was using MTT method [3- (4,5-dimethyltiazole-2-yl) -2,5 diphenyltetrazolium bromide] and then analyzed using SPSS 22. Examination of apoptosis, cell cycle, caspase 3/7 expression, ROS expression, mitochondria membrane permeability,expression of PI3K, mTOR, cyclin D1 dan p53 proteins were examined by flow cytometry method. Examination of Bcl-2 expression was examined by immunocytochemistry method. DNA fragmentation was analyzed by electrophoresis method and nuclear morphology was examined by nuclear staining method and analysis of expression PI3KCA, Akt-1, Akt-2, mTOR, EGFR, VEGFR-2, ERα and p53 genes with RT-PCR method. In vitro antioxidant activity was determined by DPPH method, ABTS and reducing power and in vivo with SOD method in female white rat which induced with doxorubicin. Determination of total phenol content by Folin-Ciocalteu method and total flavonoid content by colorimetric method.
The results from this study showed that the cytotoxic results (IC50) was 48.94 ± 0.32μg/mLfor FEAAF and 62,53 ± 0,55μg/mL for FEAPH. Ethylacetate fraction of poguntano herbs and andaliman fruits have anticancer activity through cell cycle arrest with accumulation at G0-G1 phase (57.23% and 60.48%), stimulate early apoptosis (72.75% and 70.12%) and late apoptosis (10.23% and 8.54%), increase caspase 3/7 expression (35.58% and 80.41%) and ROS expression (76,78% and 76.81%), decrease mitochondrial membrane permeability (11.68% and 19.02), reduce expression of protein Bcl-2 and cyclin D1 (45.94% dan 69.19%), increase expression of protein p53 (8.10% dan 12.83%), decrease expression of protein PI3K (2.93% dan 4.71%) and mTOR (14.03% dan 42.83%), increase of DNA fragmentation and cause exchange of nuclear morphology T47D cells. Decrease of genes expression of PI3KCA, Akt-1, Akt-2, mTOR, EGFR, VEGFR-2, ER-α and increase gene expression of p53.Cell viability in the FEAPH and FEAAFcytoprotective assay at concentrations of 100 μg/mL was 88.83 ± 2.90% and 87.03 ± 1.36%. FEAPH and FEAAF have cytoprotective activity which decrease ROS expression (62.58% to 54.63% and 58.22%). Antioxidant activity with DPPH method IC50 value (166.90 ± 0.10 and 156.48 ± 0.04 μg /mL), percent scavenging of ABTS method at 200 μg/mL concentration (35.21 ± 0.03% and 45.78 ± 0.21%), absorbance of reducing power method (0.2172 ± 0.0002 and 0.4044 ± 0.0002). The total phenol and flavonoid content of FEAPH and FEAAF (92.88 ± 0.50 mg/g and 90.41 ± 0.06 mg/g) and (84.39 ± 0.07 mg/g and 77.56 ± 0.02 mg/g). SOD levels in female white rats at dose of 300 mg/Kg BW (3.242 ± 0.086 and 5.644 ± 0.152 U/mL). Histopathologic observation of liver tissue appears to be less damage than controls. This study shows that FEAPH and FEAAFhave potential as anticancer and antioxidant where FEAAF more potential if compare to FEAPH. | en_US |
