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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."

"Latar Belakang: Endometriosis dan infertilitas memiliki keterkaitan yang sangat erat. Namun etiopatogenesis terjadinya infertilitas pada kasus endometriosis sangat beragam. Teori yang berkembang akhir-akhir ini adalah buruknya reseptivitas endometrium. Gen HOXA 11 adalah salah satu gen yang berperan dalam reseptivitas endometrium karena berkorelasi dengan penanda lain seperti Leukemia Inhibitory Factor LIF, B3integrin, dan EMX2. Teori epigenetik yang berkembang adalah terjadi hipermetilasi pada gen HOXA 11 sehingga terjadi penurunan ekspresi gen tersebut.Metode: Penelitian potong lintang ini dilakukan di RS Cipto Mangunkusumo pada Juli 2015 - Juni 2016. Subjek penelitian adalah pasien endometriosis yang terbukti secara histopatologi dengan infertilitas dan kelompok kontrol merupakan pasien non-endometriosis yang fertil. Status metilasi gen HOXA 11 dari sampel endometrium eutopik pada kedua kelompok ini diperiksa dan dibandingkan.Hasil: Enam pasien endometriosis dan enam pasien kontrol diambil sebagai subjek. Perbedaan tingkat metilasi gen HOXA 11 pada kedua kelompok ini berbeda secara signifikan dengan nilai p 0.03 dengan perbedaan rerata peningkatan kadar metilasi pada kelompok pasien endometriosis sebesar 33.Kesimpulan: Gen HOXA 11 yang berperan dalam reseptivitas endometrium mengalami hipermetilasi pada pasien dengan endometriosis dan infertilitas.

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Introduction: Endometriosis compromises infertility in some patients. They have close relationship and many etiologies have been proposed. HOXA11 has important role in window implantation because it is related to other endometrial receptivity markers such as Leukemia Inhibitory Factor LIF , B3integrin, and EMX2. Recently, many researchers found poor endometrial receptivity in endometriosis due to hyper methylation of HOXA11 gene. Therefore, this study aims to find out the HOXA11 gene profile on endometriosis patients with infertility in Indonesia.

Methods: This cross sectional study was conducted in Dr. Cipto Mangunkusumo Hospital from July 2015 June 2016. The subjects were endometriosis patients with infertility who have been confirmed histopathological. The control group was taken from non endometriosis and fertile patients. Eutopic endometrium samples were taken and examined for the methylation of HOXA11 gene.

Results: Both groups consist of six patients. The difference of methylation of HOXA 11 gene between those two groups is statistically significant p 0.03 . There was hyper methylation in endometriosis group.

Conclusion: There is a hyper methylation of HOXA 11 gene in eutopic endometrium of endometriosis patients with infertility. Thus, possibly can explain the poor endometrial receptivity in endometriosis patient and give a broad research area in epigenetic therapy of endometriosis."

"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."

"Endometriosis merupakan jaringan yang ditemukan di luar uterus dan dapat menjadi faktor penyebab terjadinya infertilitas. Endometriosis dapat terjadi pada wanita umur reproduktif dengan probabilitas sebesar 10—15%. Selain itu, endometriosis dapat terjadi pada wanita yang memiliki nyeri kronis pada pelvis dengan besar probabilitas sebesar 70%. Pertumbuhan dan perkembangan jaringan endometriosis dipengaruhi oleh autofagi. Mekanisme autofagi yang terjadi pada tubuh manusia dapat dipicu dengan keberadaan HMGB1. Selain dengan HMGB1, autofagi dapat dipicu dengan adanya ekspresi gen LC3 dan BECN1. Adanya HMGB1 berpengaruh terhadap jumlah gen LC3 dan BECN1. Penelitian dilakukan dengan menganalisis sampel jaringan endometriosis pada fase proliferasi dan sekretori menggunakan metode RT-qPCR absolut dengan kurva standard. Kurva standard dibuat dengan fragmen gen HMGB1 didapatkan nilai efisiensi sebesar 88,38% dan nilai R2 sebesar 0,99618. Kedua nilai yang telah disebutkan masuk ke dalam kisaran yang dapat diterima yaitu dalam rentang 80—110% dan diatas 0,99. Sementara itu, Uji T menunjukkan perbedaan dengan hasil yang tidak signifikan (p<0,05) namun menunjukkan kecenderungan pada rata-rata kelompok. Penelitian ini juga melakukan uji Spearman’s Rank pada HMGB1 dan BECN1 dan menunjukkan korelasi positif baik itu pada kelompok sekretori maupun proliferasi dengan hasil masing-masing 0,721 dan 0,729. Sehingga dapat disimpulkan dari penelitian yang dilakukan bahwa jumlah salinan gen HMGB1 berperan dalam mekanisme autofagi dan memengaruhi tingkat autofagi pada jaringan endometriosis.

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Endometriosis is human tissue found outside the uterus and can be a contributing factor to infertility. Endometriosis can occur in women of reproductive age with a percent probability of 10—15%. In addition, it can occur in women who have chronic pelvic pain with a 70%. The growth of endometriotic tissue can be influenced by autophagy. The mechanism of autophagy that occurs in the human body can be triggered by the presence of the HMGB1 protein. In addition to the HMGB1 protein, autophagy also regulated by the presence of LC3 and BECN1 genes. The presence of HMGB1 protein affects LC3 and BECN1. The study was conducted by analyzing tissue samples of endometriosis in the proliferative and secretory phase using the RT-qPCR absolute method with standard curve. The standard curve was made by HMGB1 gene fragment and obtained an efficiency value of 88.38% and R2 value of 0.99618. The two values ​​mentioned above fall into the acceptable range, namely in the range of 80-110% and above 0.99. Meanwhile, the results the T test showed there is no significant difference (p<0,05) but showed a tendency if it compared by the group means. Spearman's Rank test showed a positive correlation between HMGB1 and BECN1 expression in both the secretory and proliferative phase with results of 0.721 and 0.729, respectively. So, it can be concluded from the research that was conducted, it was found that the expression of HMGB1 gene mRNA in the secretory phase was higher in endometrial tissue while in the proliferative phase it was higher in endometrial tissue. In addition, it can be concluded that there is a tendency for the HMGB1 gene take parts to autophagy mechanism in endometriotic tissue.

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"ABSTRAKLatar Belakang: Endometriosis merupakan isu utama yang sering ditemukan pada wanita usia reproduksi dengan keluhan infertilitas. Endometriosis Fertility Index EFI merupakan suatu sistem staging yang sederhana, bermakna, dan bermanfaat secara klinis untuk memprediksi prognosis fertilitas pada pasien endometriosis. Penelitian ini bertujuan untuk mengetahui angka kehamilan dan hubungannya dengan skor EFI pada pasien endometriosis yang menjalani terapi pembedahan laparoskopi di Rumah Sakit Cipto Mangunkusumo dan Rumah Sakit Carolus. Metode: Penelitian ini merupakan penelitian analitik dengan metode kohort prospektif yang dilakukan di RSCM dan RS Carolus pada subjek pasien endometriosis dengan keluhan infertilitas yang dilakukan terapi pembedahan laparoskopi pada tahun 2012-2014. Skor EFI ditentukan dengan mengambil data catatan rekam medis dan video operasi berdasarkan klasifikasi menurut Adamson. Data kehamilan diambil dalam periode 2 tahun follow up. Hubungan antara skor EFI dengan kehamilan dianalisis secara bivariat. Hasil: Terkumpul 51 pasien yang dianalisis. Sebanyak 18 pasien 35,3 diketahui hamil selama durasi pemantauan 2 tahun. Insidensi kehamilan sampai dengan tahun kedua pada kelompok skor EFI 0-3, 4, 5, 6, 7-8, 9-10 beturut turut adalah 0 , 0 , 50 , 25 , 92,9 , 100 . Median skor EFI pasien yang hamil vs tidak hamil yakni: 7 5-9 vs 4 1-8 dengan nilai p

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ABSTRACTBackground Endometriosis is among the main issue related to infertility among reproductive age women. Endometriosis Fertility Index EFI is a simple staging system, significant, and has clinical benefit to predict fertility prognosis among endometriosis patients. This study aimed to know incidence of pregnancy and the relationship between Endometriosis Fertility Index EFI score and pregnancy among endometriosis patients underwent laparoscopy in Cipto Mangunkusumo and Carolus Hospitals. Method This was a cohort prospective study conducted in Cipto Mangunkusumo and Carolus Hospitals. Subjects were endometriosis patinets with infertility who underwent laparoscopic surgery at 2012 2014. EFI score was noted by medical records and procedure video, using Adamson criteria. Pregnancy data was collected with two years of follow up duration. The relationship betweem EFI score and pregnancy was analyzed. Results Fifty one patients was recruited in this study with 18 of them 35,3 konw to be pregnant in follow up. Incidence of pregnancy in two years based on EFI score 0 3, 4, 5, 6, 7 8, 9 10 were respectively 0 , 0 , 50 , 25 , 92,9 , 100 . The median EFI score of pregnant patients vs non pregnant was 7 5 9 vs 4 1 8 , p"

"Endometriosis adalah kelainan ginekologis yang ditandai dengan adanya jaringan endometrium yang tumbuh di luar uterus. Penyakit ini bersifat multifaktorial, salah satunya dipengaruhi genetik. Polimorfisme genetik gen reseptor progesteron (PR) diketahui berhubungan dengan penyakit endometriosis. Penelitian ini bertujuan untuk mengetahui hubungan antara polimorfisme gen PR rs544843047 di bagian promoter dengan endometriosis di Indonesia. Penelitian ini menggunakan desain cross sectional, dengan membandingkan 25 jaringan endometriosis dari wanita penderita endometriosis dan 21 jaringan endometrium dari wanita tanpa endometriosis. Molekul DNA dari kedua jenis jaringan diisolasi, diamplifikasi dengan menggunakan metode PCR. Analisis perubahan nukleotida pada gen PR dilakukan dengan metode sequencing. Hasil penelitian menunjukkan frekuensi genotip dan alel pada SNP gen PR rs544843047 adalah genotip TT 100% dan alel T 100%. Penelitian ini menyimpulkan bahwa tidak ada hubungan antara SNP gen PR pada rs544843047 dengan penyakit endometriosis di Indonesia.

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Endometriosis is a gynecological disorder characterized by the presence of endometrial tissues that grow outside the uterus. This disease is multifactorial cause, one of which is influenced by genetics factor, and genetic polymorphism of the Progesterone Receptor (PR) gene is known to be associated with endometriosis. The aim of this study was to determine the relationship between PR gene polymorphism rs544843047 in the promoter and endometriosis in Indonesia. A cross sectional design was used in this study, comparing 25 endometriosis tissues of women with endometriosis and 21 endometrial tissues of women without endometriosis. DNA molecules from both types of tissues were isolated, then amplified using the PCR method. While analysis of nucleotide changes in the PR gene was conducted by sequencing. The results showed that the genotypic and allelle frequencies of the PR rs544843047 SNP were 100% TT genotype and 100% T allele. This research concludes that there are no association between SNP PR gene in rs544843047 and endometriosis in Indonesia."

"Endometriosis merupakan penyakit ginekologi yang ditandai dengan pertumbuhan jaringan mirip endometrium di luar rongga uterus. Inflamasi kronik pada endometriosis memiliki peranan penting dalam memfasilitasi perkembangan kista endometriosis. Penelitian ini bertujuan untuk menganalisis efektivitas oktil galat dalam menurunkan inflamasi pada tikus Wistar model endometriosis. Sejumlah 30 ekor tikus wistar betina dibagi secara acak ke dalam tiga kelompok. Kelompok pertama dan kedua direkayasa membentuk jaringan endometriosis, sedangkan kelompok ketiga dilaparatomi sebagai kelompok kontrol negatif. Setelah dua bulan, dilakukan laparatomi kedua pada kelompok satu dan dua untuk mengevaluasi pembentukan jaringan endometriosis. Induksi oktil galat diberikan pada kelompok pertama selama satu bulan. Seluruh tikus kemudian dieuthanasia dan jaringan endometriosis kelompok pertama dan kedua, serta jaringan endometrium kelompok ketiga, diambil untuk dianalisis. Pengukuran kadar sitokin TNF-α dan IL-10 dilakukan menggunakan Luminex Multiplex Assay, sedangkan kadar COX-2 dan PGE₂ diukur menggunakan metode ELISA. Analisis beda proporsi menunjukkan bahwa pemberian oktil galat pada kelompok pertama tidak memberikan perubahan kadar TNF-α kategori tinggi yang signifikan, sedangkan kadar COX-2, PGE₂, dan IL-10 kategori tinggi teramati mengalami penurunan signifikan sebesar 22,3%, 55,6%, dan 44,5%, dibandingkan dengan kelompok kedua (p<0,05). Oktil galat diketahui efektif dalam menurunkan mediator inflamasi COX-2 dan PGE₂, serta anti-inflamasi IL-10, yang memicu perbaikan gejala klinis berupa regresi ukuran kista endometriosis.

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Endometriosis is a gynecological disease, characterized by the growth of endometrial-like tissue outside the uterine cavity. Chronic inflammation in endometriosis has an important role in facilitating the development of endometriosis cysts. The present study aimed to analyze the anti inflammatory effect of octyl gallate in endometriosis Wistar rat model. 30 female Wistar rats were divided randomly into three groups. Endometriosis induction was performed in the first and second group, while a sham operation was performed in the third group. Two months later, a second laparotomy was performed in the first and second groups to evaluate endometriosis tissue formation. Octyl gallate was administered via oral gavage to the first group for one month. All rats were sacrificed and endometriosis tissue samples were collected for further analysis. TNF-α and IL-10 levels were measured using Luminex Multiplex Assay, while COX-2 and PGE₂ levels were measured using the ELISA method. The administration of octyl gallate in the first group did not significantly effect TNF-α levels, whereas the high category of COX-2, PGE₂, and IL-10 levels were observed to experience a significant decrease up to 22.3%, 55.6%, and 44.5% compared to the second group (p<0.05). In conclusion, octyl gallate was able to supress the inflammatory mediators, COX-2 and PGE₂, along the anti-inflammatory mediators IL-10, which induced the regression of endometriosis cysts size as an improvement of clinical symptoms."

"ABSTRAKEndometriosis merupakan penyakit ginekologi ditandai dengan implantasi jaringan endometrium di luar rongga uterus, berhubungan erat dengan proses inflamasi kronis. Stres oksidatif menjadi aktivator terjadinya proses inflamasi kronis di endometriosis. Oktil galat terbukti lebih efektif menekan proses inflamasi dibandingkan asam galat dan heptil galat pada sel kultur primer endometriosis. Penelitian ini bertujuan menganalisis pengaruh oktil galat pada proses inflamasi dan stres oksidatif pada tikus Wistar model endometriosis. Tiga puluh ekor tikus Wistar dibagi menjadi tiga kelompok, yaitu kelompok uji, kontrol endometriosis dan kelompok normal. Kelompok uji dilakukan autotransplantasi lalu diberikan suspensi oktil galat dan CMC selama satu bulan. Kelompok endometriosis dilakukan autotransplantasi lalu diberikan larutan CMC selama satu bulan, sedangkan kelompok normal hanya dilakukan laparotomi. Seluruh tikus kemudian dieuthanasia, dari kelompok uji dan kontrol endometriosis diambil jaringan endometriosisnya sedangkan dari kelompok sehat diambil jaringan endometriumnya untuk dianalisis. Analisis MDA (Malondialdhyde) dan SOD (Superoxide Dismutase) dilakukan secara spektofotometri, kadar NF-ĸB dengan ELISA dan IL-1β (Interleukin-1 Beta) dengan LUMINEX. Pemberian oktil galat pada kelompok uji tidak menurunkan kadar MDA namun berpotensi menekan kondisi stres oksidatif dengan meningkatkan kadar SOD. Oktil galat terbukti menekan aktivasi NF-ĸB secara signifikan, namun tidak menekan kadar IL-1β. Oktil galat berperan sebagai antiinflamasi pada tikus Wistar model endometriosis dengan cara induksi peningkatan SOD dan hambatan langsung pada translokasi nuklear NF-ĸB.

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ABSTRACTEndometriosis is a gynecological disease characterized by the implantation of endometrial tissue outside the uterine cavity, related to the chronic inflammatory process. Oxidative stress activates the occurrence of chronic inflammatory in endometriosis. Octyl gallate is more effective in suppressing the inflammatory process than gallic acid and heptil gallate in primary endometriosis culture cells. This study aimed to analyze the effect of octyl gallate on the inflammatory process and oxidative stress in endometriosis Wistar rat models. 30 Wistar rats were divided into three groups, the test group, endometriosis control and normal groups. The test group was autotransplantated and then given a suspension of octyl galate and CMC for one month. The endometriosis group was autotransplanted and then given a CMC solution for one month, while the normal group only underwent laparotomy. All rats were then euthanized, from the test and endometriosis group the endometriosis tissue was taken while from the normal group endometrial tissue was taken for analysis. MDA and SOD were measured using spectrophotometry, NF-ĸB with ELISA and IL-1β with LUMINEX. Induction of octyl gallate in the test group did not reduce MDA levels but could potentially suppress oxidative stress conditions by increasing SOD levels. Octyl gallate significantly inhibit the NF-ĸB activation, but not suppressing IL-1β levels significantly. Octyl gallate act as anti-inflammatory agent in endometriosis Wistar rat model through the enhancement of SOD and direct inhibition to nuclear translocation of NF-ĸB."

"ABSTRAKLatar belakang. Proses inflamasi kronik dan persisten mempengaruhi tingginya rekurensi dan survival endometriosis pasca pembedahan. Hal ini menjadi permasalahan endometriosis, sehingga perlu pengembangan terapi target salah satunya yaitu asam galat. Asam galat terbukti efektif sebagai antikanker, anti tumor, anti inflamasi dan antibakterial pada beberapa cell line, namun efektifitasnya pada sel endometriosis harus dibuktikan. Tujuan. membuktikan efek asam galat dan senyawa turunannya terhadap regulasi inflamasi pada kultur primer endometriosis ditinjau dari ekspresi mRNA NF-kB, serta sekresi TNF-? dan IL-6. Metode. Sel endometriosis berasal dari jaringan endometriosis pasien yang menjalani laparaskopi, diisolasi secara enzimatis dan dikultur primer. Sel kultur diberi perlakuan asam galat, heptil dan oktil galat dengan dosis 25,6 g/mL, 51,2 g/mL dan 102,4 g/mL selama 48 jam, kemudian diinduksi dengan LPS 500 ng/mL selama 24 jam. Regulasi inflamasi dinilai dari ekspresi mRNA NF-kB dengan qRT-PCR, kadar sekresi TNF-? dan IL-6 dengan ELISA, serta inhibisi viabilitas sel dengan MTS Assay. Hasil. Setelah data dirasiokan dengan kontrol, ketiga zat signifikan menghambat viabilitas sel endometriosis p value 0,000 dengan inhibisi tertinggi pada dosis 102,4 g/mL. Terjadi penurunan ekspresi relatif NF-kB yang dirasiokan dengan kontrol dan IL-6 meskipun secara statistik tidak bermakna. Konsentrasi TNF? tidak berbeda secara bermakna p value 0,340 . Kesimpulan. Asam galat dan senyawa turunannya berpengaruh terhadap inhibisi viabilitas sel, penurunan ekspresi relatif NF-kB dan IL-6, namun tidak bermakna terhadap penekanan sitokin TNF-?. Perlu dilakukan studi lanjut untuk menilai efektifitas asam galat sebagai kandidat obat antiinflamasi pada endometriosis ditinjau aspek lain.

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ABSTRACTBackground. Endometriosis is a benign gynecological disorder characterized by the growth of the lining of the endometrium like tissue outside the uterus. The cause of the growth of endometriosis is not known well, chronic and persistent inflammatory process is suspected to be one of the pathogenesis that contributes to the high recurrence and survival endometriosis. One of the potential therapeutic agents is a gallic acid which proved effective in earlier studies as an anti cancer, anti tumor, anti inflammatory and antibacterial in several cell line. The Effectiveness of gallic acid to the endometriosis cell is a preliminary study and have not found evidence of publication yet. Object. Proving the effect of gallic acid and its derivatives on the inflammatory regulation of endometriosis primary culture study on mRNA expression of NF kB, TNF , and IL 6 secretion. Method. Endometriosis cells from Indonesian endometriosis patients tissues who had undergone laparoscopy surgery were isolated by the enzymatic reaction and primary cultured. Cultured cells treated by gallic acid and alkyl ester synthetic derivatives of the gallic acids heptyl gallate and octyl gallate each with the dosage of 25,6 g mL, 51,2 g mL, and 102.4 g mL for 48 hours and then induced by LPS 500 ng mL for 24 hours. Parameter research was assessed by qRT PCR for mRNA expression of NF kB, ELISA for the quantification of TNF and interleukin 6, and MTS assay was used to observe endometriosis cell viability. Results. After the data was rationalized with the control, three substances showed significant inhibition of endometriosis cell viability. The highest inhibition for all treatment was at doses 102,4 g mL. Overall there was an inhibition of relative expression of mRNA NF kB were rationalized to controls and suppression of IL 6 in octyl gallate groups. The concentration of TNF among the groups did not differ significantly p value 0.340 . Conclusion. Gallic acid and its derivatives have significantly inhibition effect toward cell viability, mRNA expression of NF kB, and IL 6 but have not significantly effect toward cytokine TNF . Further studies need to be conducted to assess the effectiveness of gallic acid as an anti inflammatory drug candidate toward to any pathway."

"ABSTRAKPenggunaan sel punca sebagai anti fibrosis hati cukup menjanjikan. Sel punca CD34 asal darah tali pusat sudah banyak digunakan dalam studi anti fibrosis hati. Penelitian ini menjelaskan efek ko-kultur antara sel stelata hepatik HSC LX-2 dan sel punca CD34 asal darah tali pusat dalam morfologi sel dan ekspresi TGF-?, tenascin-C dan kolagen tipe 1A1. Metode : Sel CD34 diisolasi dari sel darah tali pusat manusia yang dikriopreservasi menggunakan separasi magnet. Sel HSC LX-2 dikultur sebagai kontrol monokultur. Sebagian dipanen dan dihitung untuk dilakukan ko-kultur dengan sel CD34 dalam rasio 1:1. Ko-kultur CD34 dan LX-2 dilakukan dengan metode kultur konvensional 2D dan 3D hanging drop. Hasil monokultur dan ko-kultur dipanen pada hari ke1, 2 dan 3 dan dilakukan pewarnaan imunositokimia tenascin-C ekstraksi RNA untuk analisis kuantitatif dengan real time PCR ekspresi TGF-? dan kolagen tipe 1A1.Hasil : Hasil menunjukkan perbedaan morfologi ko-kultur 2D dan 3D hanging drop dibandingkan kontrol monokultur. Pada ko-kultur 2D terdapat mikromassa, sedangkan pada monokultur 2D tidak ada mikromassa yang terbentuk. Pada ko-kultur 3D hanging drop, terdapat spheroid yang lebih kecil hambatan pembentukan spheroid dibandingkan monokultur 3D hanging drop. Sel CD34 memiliki efek direk terhadap aktivitas sinyal sel stelata hepatik dengan adanya kecenderungan penurunan ekspresi TGF-?. Analisis imunositokimia tenascin-C dalam mikromassa dan spheroid masih perlu dioptimasi. Ko-kultur 2D dan 3D hanging drop method sel punca CD34 asal darah tali pusat dan sel stelata hepatik memiliki efek terhadap penurunan ekspresi kolagen tipe 1A1.Kesimpulan : Sel punca CD34 asal darah tali pusat memiliki efek direk terhadap morfologi sel, inhibisi aktivitas sel stelata hepatik LX-2 yang ditandai dengan penurunan ekspresi TGF-beta dan inhibisi deposisi matriks ekstrasel yang ditandai penurunan ekspresi kolagen tipe 1A1.Kata kunci: sel punca asal darah tali pusat CD34 , sel stelata hepatik, liver fibrosis, TGF-beta, tenascin-C, kolagen 1A1.

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ABSTRACTBackground The development of stem cell therapy antifibrotik placing as one of the promising therapy. Umbilical cord blood CD34 stem cells has been widely used in the study antifibrosis. This study describes the effect of co culture between hepatic stellate cells HSC LX 2 and umbilical cord blood CD34 stem cells on cell morphology and expression of TGF , tenascin C and collagen type 1A1.Method CD34 cells were isolated from thawed cryopreserved human umbilical cord blood cells using magnetic separation. LX 2 cells culture were harvested and counted. CD34 and LX 2 cells were mixed in suspension with 1 1 ratio v v . Cell suspension divided into 2 sets 2D co culture plated in standard well plate and 3D co culture as hanging drops. LX 2 monoculture, CD34 dan LX 2 coculture were harvested on day 1, 2 and 3 as sample for further analysis. Tenascin C expression was analysed by imunocytochemistry techniques. TGF Beta and collagen type 1A1 expression was analysed by qPCR.Result The result showed different morphology between co culture and monoculture on 2D and 3D hanging drop. The 2D co culture showed micromass formation, instead of no micromass formation on monoculture. The 3D hanging drop showed smaller spheroid formation spheroid formation inhibition compared with monoculture. CD34 cells showed direct effect on hepatic stellate cell signalling activity represented by the decrease in TGF beta expression, inhibition of extracellular matrix deposition represented by a decrease in Collagen type 1A1 expression.Conclusion UCB CD34 cells showed direct effect on cell morphology, inhibition of hepatic stellate cell LX 2 activity represented by a decrease in TGF beta expression, inhibition of extracellular matrix deposition represented by a decrease in collagen type 1A1 expression. "