Skripsi

Comparison between Apoptotic Induced by Gallic Acid and Alkyl Gallate Derivatives on Endometriosis Cells In Vitro = Perbandingan antara Apoptosis Dipicu oleh Asam Galat dengan Derivat Alkil Galat Pada Sel Endometriosis In Vitro.

Research into endometriosis falls behind despite increasing publication for the last five years, contributing to lack of non-invasive treatments and 20% decrease in quality of life. Since gallic acid usage as anti-inflammatory agent has been elucidated in cancer cells, this study serves to investigate the potential of gallic acid as an apoptotic inducer in endometriosis cells. Primary culture of endometriosis was derived from patients who had laparascopy via enzymatic technique. In vitro endometriosis cells were treated with three dosages of 25.6 µg/mL, 51.2 µg/mL and 102.4 µg/mL of gallic acid, heptyl gallate, and octyl gallate for 48 hours. Determination of quality and quantification of early, late, viable, and necrotic cells was done using confocal fluorescence with acridine orange/ethidium bromide staining of at least 100 cells per sample. Control samples showed 63.8% cell underwent apoptosis. Gallic acid, heptyl gallate, and octyl gallate showed different inhibition pattern. Apoptosis after gallic acid treatment decreased from 90.1% to 79.2% as the dose is increased. On the contrary, 51.2 µg/mL heptyl induce 92.5% apoptosis, while octyl showed most apoptosis at 93.1% This study exhibited apoptotic inductor effect of octyl gallate, followed by heptyl gallate and gallic acid and their potency as treatment for endometriosis.
Keywords: Endometrial cells, endometriosis, gallic acid, heptyl gallate, octyl gallate, apoptosis, confocal fluorescence, AO/EB staining


Walaupun publikasi mengenai endometriosis meningkat dalam lima tahun terakhir, penanganan dan terapi definitif untuk endometriosis masih belum ditemukan, menyebabkan 20% penurunan kualitas hidup pada pasien endometriosis. Asam galat telah digunakan pada sel kanker sehingga mendorong peneliti untuk menyelidiki potensi asam galat sebagai pemicu apoptosis pada sel endometriosis. Kultur primer didapatkan dengan teknik enzimatik dari pasien yang telah menjalani laparoskopi. Sel endometriosis in vitro diberi perlakukan dengan asam galat, heptil galat, dan oktil galat dengan dosis 25.6 µg/mL, 51.2 µg/mL, dan 102.4 µg/mL selama 48 jam. Kuantifikasi dan penentuan kualitas sel dilakukan menggunakan mikroskop konfocal fluoresens dan pewarnaan Acridine Orange/Ethidium Bromide (AO/EB) dengan observasi sel yang mengalami apoptosis awal, apoptosis akhir, nekrosis, dan sel hidup. Sampel kontrol menunjukkan 63.8% sel mengalami apoptosis. Apoptosis setelah pemberian asam galat menurun dari 90,1% menjadi 79,2% dengan peningkatan dosis. Sebaliknya, 51,2 6 µg/mL heptyl meningkatkan apoptosis menjadi 92.5%, sedangkan oktil 51,2 6 µg/mL menunjukkan apoptosis terbesar dengan 93.1%. Penelitian ini menunjukkan efek apoptosis oktil galat diikuti oleh heptil galat dan asam galat dan potensi penggunaan oktil galat sebagai terapi endometriosis.
Kata Kunci: Sel endometrium, endometriosis, asam galat, heptil galat, oktil galat, apoptosis, konfokal fluoresens, pewarnaan AO/EB