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"ABSTRAKLatar belakang. Potensi terjadinya kekambuhan paska pengobatan endometriosisdengan terapi hormonal dan pembedahan konservatif masih terjadi sekitar 11-32dalam waktu 1-5 tahun. Salah satu faktor pemicunya adalah proses inflamasi kronikyang merangsang peningkatan sitokin proinflamasi dalam rongga peritoneum, sehinggaperlu pengembangan terapi baru. Heptil galat dan oktil galat merupakan senyawaturunan asam galat yang berpotensi menekan proliferasi beberapa jenis sel kanker.Penelitian kami sebelumnya membuktikan oktil galat dapat menekan ekspresi mRNANFkB yang merupakan faktor transkripsi aktivasi jalur proinflamasi, serta dapatmenekan proliferasi sel endometriosis in vitro. Saat ini kami ingin menganalisisaktivitas heptil galat dan oktil galat terhadap protein target NFkB melalui teknik insilicodocking dan efeknya terhadap regulasi sitokin proinflamasi IL-1, COX-2, TGF-1 dan IL-10 pada kultur primer sel endometriosis.Metode. In silico docking heptil galat dan oktil galat terhadap protein target NFkBmelalui teknik bioinformatika. Sel endometriosis dari jaringan primer pasien diisolasisecara enzimatis dan dikultur, kemudian diberi perlakuan heptil dan oktil galat dengan 2macam dosis (51,2 μg/mL dan 102,4 μg/mL) selama 48 jam, dilanjutkan induksi LPS 10ng/mL selama 24 jam. Kelompok kontrol positif hanya diinduksi LPS tanpa perlakuan,dan kontrol negatif tanpa perlakuan dan LPS. Regulasi inflamasi dinilai dari tingkatkadar sitokin IL-1, COX-2, TGF-1 dan IL-10 dengan teknik ELISA.Hasil. Analisis in-silico docking protein NFkB menunjukan nilai ikatan energi oktilgalat lebih tinggi (-7,98 kkal/mol) dibandingkan heptil galat (-7,68 kkal/mol) dan asamgalat (-7,66 kkal/mol). Terjadi penurunan kadar sitokin COX-2 secara signifikan(p<0,03) pada kelompok perlakuan dibandingkan dengan kontrol positif, begitu jugadengan sitokin IL-1 dan IL-10 cenderung menurun (p>0,05). Sedangkan kadar sitokinTGF-1 mengalami kenaikan pada kelompok perlakuan dibandingkan kontrol positifmeskipun kurang bermakna secara statistik (p>0,05).Kesimpulan. Melalui jalur NF-kB sebagai regulator inflamasi, baik oktil galat danheptil galat terbukti dapat menekan produksi sitokin proinflamasi COX2 dan IL-1serta meningkatkan sitokin TGF-1 dan menurunkan sitokin IL-10 sehingga berpotensisebagai bahan terapi tambahan pada endometriosis.

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ABSTRACTBackground: The potential for relapse post endometriosis treatment with hormonaltherapy and conservative surgery still occurs around 11-32 within 1-5 years. One ofthe trigger factors is a chronic inflammatory process that stimulates an increaseproinflammatory cytokines in the peritoneal cavity, so needed the development of newtherapies. Heptyl galate and octyl galate are gallic acid derivatives which have thepotential to suppress the proliferation of several types cancer cells. Our previousresearch proved that octyl galate can suppress the expression of NFkB mRNA which isa proinflammatory activation transcription factor, and can suppress endometriosis cellproliferation in vitro. We currently want to analyze the activity of heptyl galates andoctyl galates against the NFκB target protein through in-silico docking techniques andtheir effects on the regulation of proinflammatory cytokines IL-1, COX-2, TGF-1 andIL-10 in primary cultures of endometriosis cells.Method: In silico docking heptyl and octyl galates against the NFkB target proteinsthrough bioinformatics techniques. Endometriosis cells from primary tissue wereenzymatically isolated and cultured, then given heptyl and octyl gallate treatment with 2doses (51.2 μg/mL and 102,4 μg/mL) for 48 hours, continued induction of 10 ng / mLLPS for 24 hours. The positive control group only induced LPS without treatment, andnegative treatment without treatment and LPS. Inflammatory regulation was assessedfrom levels of cytokines IL-1, COX-2, TGF-1 dan IL-10 with ELISA techniques.Results: In-silico docking analysis of the NFkB gene showed higher energy bondingvalues in octyl galate (-7,98 kcal / mol) than heptyl galate (-7,68 kcal / mol) and gallicacid (-7,66 kcal / mol). Significantly decreased levels of COX-2 cytokine (p <0,03) inthe treatment group compared with positive controls, so also the cytokines of IL-1 andIL-10 tended to decrease (p> 0,05). Whereas the levels of cytokine TGF-1 experiencedan increase in the treatment group compared to the positive control although it was lessstatistically significant (p> 0,05).Conclusion: Through the NFkB pathway as an inflammatory regulator, both octylgalates and heptyl galates have been shown to suppress the production ofproinflammatory cytokines COX2 and IL-1, as well as increase TGF-1 cytokines andreduce IL-10 cytokines so that they have the potential to be additional therapeuticagents in endometriosis."

"Asam galat dan oktil galat adalah senyawa yang telah banyak digunakan di berbagai industri. Asam galat dikenal memiliki manfaat terapeutik potensial sebagai antibakteri, antivirus, antijamur, anti-inflamasi, dan anti kanker. Mirip dengan salah satu turunannya, octyl gallate. Turunan asam galat ini yang memiliki 8 rantai karbon alkil, berfungsi sebagai antibakteri, antifugal, dan antioksidan kuat. Tingginya kasus resistensi Salmonella thypi dan Streptococcus pneumoniae memberi peluang untuk asam galat dan oktil galat sebagai alternatif antibiotik baru. Pengujian dilakukan dengan metode difusi cakram diikuti dengan mengukur zona hambat yang terbentuk. Salmonella thypi dan Streptococcus pneumoniae diuji dengan asam galat dan oktil gallate dengan konsentrasi 16 mg / L, 32 mg / L, 64 mg / L, 128 mg / L, 128 mg / L, 256 mg / L, 512 mg / L, dan 1024 mg / L. Setiap tes diulang tiga kali (triplo). Dalam tes ini, gentamisin disc 10 μg digunakan sebagai kontrol positif. Hasil penelitian ini dinyatakan dalam satuan mm berdasarkan zona penghambatan setiap senyawa. Dari hasil pengujian menggunakan 7 konsentrasi yang berbeda, asam galat dan oktil galat belum mampu menghambat aktivitas Salmonella thypi dan Streptococcus pneumoniae, karena zona hambat yang diamati adalah 0 mm hingga konsentrasi 1024 mg / L.

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Gallic acid and octyl gallate are compounds that have been widely used in various industries. Gallic acid is known to have potential therapeutic benefits as antibacterial, antiviral, antifungal, anti-inflammatory, and anti-cancer. Similar to one of its derivatives, octyl gallate. This gallic acid derivative which has 8 alkyl carbon chains, functions as an antibacterial, antifugal, and strong antioxidant. High cases of resistance of Salmonella thypi and Streptococcus pneumoniae provide opportunities for gallic acid and octyl gallate as alternatives to new antibiotics. Tests carried out by the method of disk diffusion followed by measuring the zone of inhibition formed. Salmonella thypi and Streptococcus pneumoniae were tested with gallic acid and octyl gallate at concentrations of 16 mg / L, 32 mg / L, 64 mg / L, 128 mg / L, 128 mg / L, 256 mg / L, 256 mg / L, 512 mg / L, and 1024 mg / L. Each test was repeated three times (triplo). In this test, a 10 μg gentamicin disc was used as a positive control. The results of this study are expressed in units of mm based on the zone of inhibition of each compound. From the test results using 7 different concentrations, gallic acid and octyl gallate have not been able to inhibit the activity of Salmonella thypi and Streptococcus pneumoniae, because the inhibited zone observed was 0 mm to a concentration of 1024 mg / L."

"Latar Belakang. Endometriosis ditandai dengan pertumbuhan jaringan endometrium di luar uterus, salah satunya disebabkan oleh disregulasi apoptosis sel yang memicu ketahanan sel ektopik. Asam galat dan turunannya pada beberapa penelitian mampu menghambat karsinogenesis pada beberapa cell line kanker. Penelitian kami terdahulu membuktikan asam galat dan turunannya dapat menekan proliferasi sel dan meningkatkan apoptosis sel endometriosis in vitro, namun efeknya terhadap mekanisme jalur apoptosis instrinsik belum di buktikan. Metode. Sel endometriosis berasal dari jaringan endometrium pasien laparaskopi, diisolasi secara enzimatis dan dikultur primer. Sel kultur diberi perlakuan asam galat, heptil galat, oktil galat dengan dosis 51,2 μg/ml, 102,4 μg/ml dan 153,6 μg/ml selama 48 jam, dilanjutkan induksi 10 ng/ml LPS selama 24 jam . Kelompok kontrol hanya di induksi LPS tanpa perlakuan. Ekspresi relatif mRNA Bax, Bcl-2, dan Caspase-3 dinilai dengan qRT-PCR. Hasil. Peningkatan tertinggi ekspresi mRNA Bax dan penekanan tertinggi ekspresi mRNA Bcl-2 pada oktil galat dosis 153,6 μg/ml. Peningkatan ekspresi mRNA Bax, dan penurunan ekspresi mRNA Bcl-2 akan di ikuti dengan peningkatan ekspresi mRNA Caspase-3. Secara statistik tidak terdapat perbedaan bermakna antara kelompok perlakuan dengan ekspresi mRNA Bax, Bcl-2, dan Caspase-3 (p > 0,05). Kesimpulan. Oktil galat, asam galat, dan heptil galat memiliki efek potensial pada mekanisme apoptosis intrinsik.

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Background. Endomeriosis characterized by the presence of extrauterine endometrial tissue, one of which caused by disregulation of apoptosis that contribute of endometrial ectopic survival. Our previous research has proven that gallic acid and its derivatives can suppress proliferation and induce apoptosis endometriosis cell in vitro. However, the effect of gallic acid and its derivatives on apoptosis intrinsic pathway mechanism is not proven yet. Method. Endometriosis cell from endometriosis patiens who had undergone laparascopy surgery were isolated by enzimatic reaction and primary cultured. Cultured cells treated by gallic acid, heptyl gallate and octyl gallate each with dosage 51.2 μg/ml, 102.4 μg/ml, 153.6μg/ml for 48 hours, than induced by LPS 10 ng/ml for 24 hours. Parameter research was assessed by qRT-PCR for mRNA expression of Bax, Bcl-2, Caspase-3. Result. Octyl gallate showed more effect to induce apoptosis intrinsic . Endometriosis cell were treated with octyl gallate shown increases of Bax and Caspase3 mRNA expression than decrease of Bcl-2 mRNA expression. Statistically, mean differences are not significant between treatment groups and mRNA expression (p > 0.05). Conclusion. This study exhibited that octyl gallate has a more potential effect on apoptosis intrinsic in endometriosis cell cultures followed by gallic acid and heptyl gallate and their potency as treatment for endometriosis."