ABSTRAK
Latar 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.
ABSTRACT
Background: 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. |
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