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Miftah Irramah

Ekspresi connexin43 dan caspase 3 pada kardiomiosit ventrikel kiri tikus overtraining = Connexin43 and caspase 3 expression in left ventricle cardiomyocytes of overtrained rats

"Latar belakang : Overtraining berdampak buruk terhadap kesehatan karena dapat menyebabkan kematian mendadak pada atlet muda. Berdasarkan data epidemiologi ditemukan bahwa kejadian kematian mendadak (suddent cardiac death) pada atlet muda, penyebab paling banyak adalah gangguan kardiovaskular. Tubuh melakukan adaptasi terhadap beban berlebih, berupa remodelling (morfologi dan elektrofisiologi). Remodeling elektrofisiologis yaitu perubahan pada gap junction, berupa perubahan ekspresi Cx43 yang yang mengakibatkan gangguan penghantaran konduksi listrik. Selama latihan fisik dapat terbentuk ROS yang akan menginduksi permeabilitas mitokondria sehingga terjadi kebocoran sitokrom c, selanjutnya akan mengaktifkan kaskade apoptosis.

Metode : Penelitian ini dilakukan pada 6 jaringan kardiomiosit tikus Wistar kelompok kontrol dan overtraining. Ekspresi Cx43 dan caspase-3 diamati melalui pulasan imunohistokimia dan diukur dengan image J.

Hasil : Hasil penelitian ini menunjukkan peningkatan bermakna pada ekspresi Cx43 total overtraining (43644.57±27711.03) dibandingkan kelompok kontrol (13002.37±3705.41). Tidak ditemukan perbedaan bermakna ekspresi caspase-3 pada kedua kelompok meskipun diperoleh hasil lebih tinggi pada kelompok overtraining (14.15%±10.54%) dibandingkan kelompok kontrol (2,63%±3.56%).

Kesimpulan : Overtraining meningkatkan ekspresi Cx43 total tetapi tidak terbukti meningkatkan caspase-3 pada kardiomiosit ventrikel kiri tikus.

Background: overtraining has bad effect for health, overtraining can cause sudden death in young athlete, reports of sudden death incidences in young athlete claim that cardiovascular disease is the cause. The heart can face the excess load by remodeling as it?s adaptation mechanism. There is 2 type remodeling, morphology and electrophysiology. Remodeling electrophysiology is a change on Cx43 expression which can interfere the heart?s electrical conduction. Free radical which formed from physical exercise can induce mitochondrial permeability that lead leakage of cytochrome c, so that so that activate the apoptosis cascade.
Methods: This study conducted on 12 Wistar rat?s cardiomyocytes tissue that divided into control and overtraining group. Cx43 expression and caspase-3 was observed through immunohistochemical staining and measured by image J.
Results: There was significant increase in the expression of Cx43 total overtraining (43644.57 ± 27711.03) compared to the control group (13002.37 ± 3705.41). Found no significant differences in the expression of caspase-3 in both groups although the result was higher in the group of overtraining (14,15% ± 10,54%) compared to the control group (2,63% ± 3,56%).
Conclusion: Overtraining increase total Cx43 expression but not proven to increase caspase-3 in the rat left ventricular cardiomyocytes."

Depok: Fakultas Kedokteran Universitas Indonesia, 2016

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Yudhistirawaty

Efek Pemberian Kinin Topikal Terhadap Pertumbuhan Rambut Mencit Model Alopesia Androgenetik yang DiinduksiTestosteron: Kajian docking in silico, enzim 5α reduktase, prostaglandin E2, malondialdehid, kaspase-3, dan histologi = The Effect of Topical Administration Qunine on Hair Growth in Testosterone-induced Alopecia Mice. in silico docking, 5α reductase, prostaglandyn E2, malondyaldehide, caspase-3, histologi study

"Latar Belakang: Alopesia androgenetik (AAG), merupakan kelainan rambut yang mengganggu secara psikososial bagi sebagian besar lelaki. Pengobatan yang ada saat ini masih terbatas. Kinin secara empirik telah digunakan sebagai zat penumbuh rambut. Namun belum ada penelitian untuk menilai mekanisme kerja dan efektivitas kinin topikal terhadap pertumbuhan rambut.

Metode: Kajian in silico molecular docking dan molecular dynamic simulation dilakukan untuk menilai afinitas kinin terhadap enzim 5α-reduktase dengan kontrol finasterid. Uji hambatan kinin terhadap 5α-reduktase secara in vitro dinyatakan sebagai nilai IC50 kinin, dengan finasterid sebagai pembanding. Pada percobaan in vivo, 28 ekor mencit C57BL/6 dibagi secara acak menjadi 7 kelompok terdiri dari 4 ekor. Kelompok (A) testosteron (alopesia),yang di suntik 1 mg testosteron secara subkutan,(B) testosteron+finasterid 2%(C) kelompok normal, dan kelompok (D-G) testosteron+kinin 0,5%,1%,1,5% dan 2 %. Dioles bahan uji sesuai kelompok setiap hari selama 28 hari. Luas dan panjang rambut dinilai secara visual setiap minggu. Pemeriksaan histologi (morfologi dan kepadatan folikel rambut), pemeriksaan imunohistokimia kaspase-3, pemeriksaan ELISA PGE2 dan pemeriksaan MDA dilakukan setelah 28 hari. Hasil : Hasil in silico menunjukkan kinin mempunyai afinitas yang cukup baik terhadap 5β-reduktase, meskipun lebih rendah dibanding finasterid (ΔG kinin -31,67 kj/mol dan ΔG finasterid -42,89 kj/mol dari hasil penambatan molekuler; serta ΔG kinin -6,32±12,84 kj/mol dan ΔG finasterid -11,17±18,92 kj/mol dari hasil simulasi dinamika molekuler. Hasil pengukuran hambatan enzim 5α-reduktase didapatkan IC50 kinin 10,6 ±1,40 uM dan IC50 finasterid 0,623 ± 0,14 nM. Luas area pertumbuhan rambut semua kelompok perlakuan kinin lebih luas dibandingkan kelompok alopesia dan berbeda bermakna pada pada minggu ke-3 dan ke-4. Gambaran histopatologi morfologi rambut semua kelompok didominasi fase anagen . Rasio anagen: telogen kelompok perlakuan kinin lebih tinggi dibandingkan kelompok alopesia. Kepadatan folikel rambut kelompok perlakuan kinin lebih tinggi secara bermakna daripada kelompok alopesia. Kinin 0,5%, 1%, dan finasterid menurunkan ekspresi kaspase-3. PGE2 meningkat pada semua kelompok perlakuan kinin, tertinggi pada kinin 2%. dan berbeda bermakna terhadap kelompok alopesia. Pemberian kinin tidak mampu menurunkan kadar MDA.

Kesimpulan: Pemberian kinin pada mencit jantan yang diinduksi testosteron mempunyai efek menumbuhkan rambut, meningkatkan kepadatan folikel rambut. Kinin menghambat enzim 5α-reduktase, meningkatkan kadar PGE2, menurunkan ekspresi Kaspase-3, tetapi tidak menurunkan MDA. Kinin potensial dikembangkan sebagai penumbuh rambut.

Background: Androgenetic alopecia (AGA), is a hair disease which lead to negative psychological effects in most of its sufferers. Currently, treatments of AGA are limited to topical minoxidil and oral finasterid, which possess many side effects. Quinine had empirically used as herbal hair grower, but its mechanism of action and effectiveness are not studied yet.

Methods: Molecular docking was done to analyse the inhibition affinity of the 5β-reductase enzyme of both quinine and finasteride as control. Further, an in vitro test for inhibition of 5α-reductase was carried out by measuring the IC50 on both compounds. The experimental study used male C57BL/6 mice aged 7 weeks. Mice were randomly divided into 7 groups of 4 animals, namely (A) testosterone (alopecia group) which was performed subcutaneous injection of 1 mg testosterone on the back of mice (B) testosterone+2% finasterid, (C) normal group, (D-G) testosterone+0.5%, 1% , 1.5% and 2% quinine. Application of the test material on the back of the mice was carried out daily for 28 days. Hair growth assessment was done visually by assessing hair growth area every week. Histologic examination ( hair morphology and density of hair follicles), immunohistochemical examination of caspase-3, ELISA examination for PGE2, and lipid peroxidase by MDA examination were carried out after 28 days.

Results: The results of molecular docking showed that quinine had a good affinity for 5β-reductase, but it was lower than finasterid (ΔG quinine = -31.67 kJ/mol vs ΔG finasterid = 42.89 kJ/mol and from dynamic simulation ΔG quinine -6,32±12,84 kj/mol and ΔG finasteride -11,17±18,92 kj/mol). The IC50 of quinine was 10.6 ± 1.40 uM, while finasterid was 0.623 ± 0.14 nM.The area of hair growth in the quinine treatment group was wider than the alopecia group and showed significance on the 3rd and 4th week. Histopathological features all of the groups was dominated by the anagen phase. The anagen:telogen ratio of the alopecia group was lower than that of the quinine group. The density of hair follicles in the treatment group and the alopecia group was statistically significant.The administration of 0.5% and 1% quinine, as well as finasterid, decreased the expression of caspase-3. PGE2 was increased in the quinine treatment group and significant compared to the alopecia group. MDA levels were higher in quinine compared to alopecia group.

Conclusion: Quinine was efficacious in promoting hair growth in AGA mouse models. It is inhibited 5α-reductase enzyme, increasing PGE2, decreasing caspase-3. However it was unable to suppress MDA levels. Hence, topical quinine has the potential to be further developed into a herbal medicine for hair growth."

Jakarta: Fakultas Kedokteran Universitas Indonesia, 2020

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Aditya Dwi Septiawan

Analisis ekspresi protein zip 1 dan caspase-3 pada adenokarsinoma prostat = Analysis expression of zip1 and caspase-3 protein in adenocarsinoma prostate

"[Pendahuluan: Proses karsinogenesis adenokarsinoma prostat terjadi akibat disregulasi kadar zinc dalam sel. Molekul zinc intrasel berperan dalam metabolisme aerob mitokondria dan induksi apoptosis. Penyerapan zinc diatur oleh protein ZIP1, berperan meningkatkan kandungan zinc sitoplasmik intrasel dengan membawa zinc dari cairan ekstrasel. Kadar zinc yang tinggi dan ekspresi protein ZIP1 banyak ditemukan pada epitel prostat normal, sedangkan pada kanker prostat ditemukan sedikit atau tidak ada ekspresi protein ZIP1. Penurunan ekspresi ZIP1 diduga dapat menghambat apoptosis, serta memacu perkembangan adenokarsinoma prostat. Penelitian ini bertujuan menganalisis korelasi ekspresi protein ZIP1 dan Caspase- 3 pada jaringan adenokarsinoma prostat berdasarkan Gleason score yang berbeda. Metode: Desain studi analitik retrospektif dengan desain potong lintang. Sampel penelitian ini adalah 31 sediaan blok parafin adenokarsinoma prostat yang memenuhi kriteria inklusi. Sediaan dipulas menggunakan teknik imunohistokimia untuk mengetahui ekspresi protein ZIP1 dan caspase-3. Ekspresi protein pada pulasan slide dihitung menggunakan program imageJ. Gleason score sebagai data sekunder yang didapatkan dari laporan kasus. Korelasi ekspresi kedua protein berdasarkan Gleason score dianalisis dengan uji korelasi Pearson menggunakan SPSS 11.5. Hasil: Rerata positivitas ekspresi ZIP1 pada adenokarsinoma prostate adalah 35% dan rerata positivitas caspase-3 adalah 33%. Terdapat korelasi positif bermakna antara ekspresi ZIP1 dan caspase-3 (r = 0.379 , p = 0,018). Terdapat korelasi positif antara ekspresi ZIP1 dan caspase-3 pada kelompok intermediate grade (r = 0.73, p = 0.01) dan korelasi lemah tidak bermakna pada kelompok high grade (r = 0.04, p = 0.48). Kesimpulan: Terdapat korelasi positif antara ekspresi ZIP1 dan ekspresi caspase- 3 pada adenokarsinoma prostat.

, Introduction: Carcinogenesis of adenocarcinoma of the prostate occurs due to dysregulation of zinc level within the cells. Intracellular zinc molecules contributes to mitochondrial aerobic metabolism. Its influx is regulated by a transporter protein ZIP1, whose non-presence is predicted to inhibit apoptosis, thus leads to the development of prostate adenocarcinoma. This study was aimed to analyze the correlation of ZIP1 and Caspase-3 expression in prostate adenocarcinoma with respect to its grading as represented by Gleason Score. Methods: This was a cross-sectional, retrospective analytical study on 31 formalyn-fixed, paraffin-embedded tissue that meet inclusion criteria. The specimen was stained using immunohistochemical technique for ZIP1 and Caspase-3. Protein expression of each case were counted using ImageJ analysis. Gleason score were acquired as secondary data from the cases’ reports. The correlation of their expression with respect of Gleason score were analyzed with Pearson’s correlation using SPSS 11.5.
Results: Mean expression level of ZIP1 and Caspase-3 in prostate adenocarcinoma were 35% and 33%, respectively. There was a significantly positive correlation between ZIP1 and Caspase-3 expression (r=0.379; p=0.018). However, their correlation was stronger in intermediate-grade group (r=0.73; p=0.01) and the correlation was much weaker in high-grade group (r=0.04; p=0.48).
Conclusion: There was a positive correlation between ZIP1 and caspase-3 expression in adenocarcinoma prostate.]"

Jakarta: Fakultas Kedokteran Universitas Indonesia, 2014

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Feriandri Utomo

Analisis ekspresi transporter zink (ZnT-1) sebagai faktor prognosis adenokarsinoma prostat = Expression analysis of zinc transporter (ZnT-1) as prognostic factor of prostate adenocarcinoma

"[ABSTRAK

Pendahuluan: Penurunan kadar Zink (Zn) pada prostat berkorelasi dengan peningkatan skor Gleason adenokarsinoma prostat dan menyebabkan rendahnya Caspase-3 sebagai eksekutor apoptosis. Tingkat ekspresi transporter Zn pada sel tumor prostat berhubungan dengan tingkat keganasannya. ZnT-1 merupakan transporter Zn ke luar sel prostat, sedangkan ZIP-1 merupakan transporter Zn ke dalam sel prostat. Ekspresi ZIP-1 turun pada adenokarsinoma prostat. Korelasi ZnT-1, ZIP-1 dan Caspase-3 diduga berpengaruh dalam karsinogenesis prostat, sehingga berpotensi untuk menjadi faktor prognosis adenokarsinoma prostat. Tujuan: Mempelajari korelasi ekspresi ZnT-1 dan aktivasi Caspase-3 terhadap skor Gleason, menganalisis korelasi ekspresi ZIP-1, ZnT-1 dan aktivasi Caspase- 3, serta mengetahui profil ekspresi ZnT-1 pada jaringan adenokarsinoma prostat yang berbeda skor Gleason, dibandingkan dengan jaringan Benign Prostatic Hyperplasia (BPH),

Desain: Studi retrospektif analitik potong lintang. Metode: Sampel penelitian ini adalah 31 blok parafin prostat yang dikelompokkan menjadi BPH, adenokarsinoma prostat skor Gleason ≤ 7 dan skor Gleason > 7. Ekspresi ZnT-1 dinilai dengan pulasan imunohistokimia. Data ekspresi ZIP-1 dan Caspase-3 merupakan data sekunder dari penelitian Septiawan et al. Hasil: Ekspresi ZnT-1 berkorelasi dengan skor Gleason adenokarsinoma prostat. Ekspresi ZIP-1 berkorelasi dengan aktivasi Caspase-3 pada adenokarsinoma prostat dan adenokarsinoma prostat skor Gleason ≤ 7. Ekspresi ZnT-1 berkorelasi dengan ekspresi ZIP-1 pada adenokarsinoma prostat skor Gleason > 7. Ekspresi ZIP-1 berkorelasi kuat dengan aktivasi Caspase-3 pada adenokarsinoma prostat skor Gleason 8. Ekspresi ZnT-1 pada adenokarsinoma prostat skor Gleason > 7 lebih rendah dibandingkan pada skor Gleason ≤ 7, tetapi tidak dapat dianalisis kemaknaan perbedaannya dengan BPH karena hanya diperoleh 1 sampel BPH pada penelitian ini. Kesimpulan: Transporter Zn berpotensi untuk menjadi faktor prognosis adenokarsinoma prostat.

ABSTRACT

Background: Decreased levels of Zinc ( Zn ) in the prostate correlates with an increase in the grade Gleason score of prostate adenocarcinoma and decrease in Caspase-3 as apoptosis executor. Zn transporter expression levels in prostate tumor cells associates with the level of malignancy. The ZnT-1 is Zn exporter, while the ZIP-1 is Zn importer of prostate cells. ZIP-1 expression drops on adenocarcinoma prostate. Correlation ZnT-1, ZIP-1 and Caspase-3 allegedly influential in prostate carcinogenesis and potential to be prognostic factor of prostate adenocarcinoma.

Objective: To analyze the correlation ZnT-1 and Caspase-3 activation of the gleason score, to analyze the correlation of the expression of ZIP-1, ZnT-1 and Caspase-3 activation in adenocarcinoma prostate, and to study the profile expression of ZnT-1 in prostate adenocarcinoma with different grading of Gleason scores, compared with benign prostatic hyperplasia (BPH),

Design: A cross-sectional retrospective study analytic . Methodology: The sample is 31 paraffin blocks were grouped into BPH, prostate adenocarcinoma Gleason scored ≤ 7 and prostate adenocarcinoma Gleason scored > 7. Samples are analyzed expression of ZnT-1 by immunohistochemical staining. ZIP-1 and Caspase-3 expression is secondary data of Septiawan et al?s immunohistochemical staining. Results: ZnT-1 expression correlated with gleason score. ZIP-1 correlated with the activation of Caspase-3 in prostate adenocarcinoma and prostate adenocarcinoma Gleason score ≤ 7. ZnT-1 correlated with ZIP-1 in prostate adenocarcinoma Gleason score > 7. ZnT-1 expression in prostate adenocarcinoma Gleason scored > 7 was lower than prostate adenocarcinoma Gleason score ≤ 7, but could not be analyzed the difference significance with BPH because there was only 1 BPH sample in this research. Conclusion: Zn transporters have the potential to be a prognostic factor of prostate adenocarcinoma.;Background: Decreased levels of Zinc ( Zn ) in the prostate correlates with an increase in the grade Gleason score of prostate adenocarcinoma and decrease in Caspase-3 as apoptosis executor. Zn transporter expression levels in prostate tumor cells associates with the level of malignancy. The ZnT-1 is Zn exporter, while the ZIP-1 is Zn importer of prostate cells. ZIP-1 expression drops on adenocarcinoma prostate. Correlation ZnT-1, ZIP-1 and Caspase-3 allegedly influential in prostate carcinogenesis and potential to be prognostic factor of prostate adenocarcinoma.

Design: A cross-sectional retrospective study analytic . Methodology: The sample is 31 paraffin blocks were grouped into BPH, prostate adenocarcinoma Gleason scored ≤ 7 and prostate adenocarcinoma Gleason scored > 7. Samples are analyzed expression of ZnT-1 by immunohistochemical staining. ZIP-1 and Caspase-3 expression is secondary data of Septiawan et al’s immunohistochemical staining. Results: ZnT-1 expression correlated with gleason score. ZIP-1 correlated with the activation of Caspase-3 in prostate adenocarcinoma and prostate adenocarcinoma Gleason score ≤ 7. ZnT-1 correlated with ZIP-1 in prostate adenocarcinoma Gleason score > 7. ZnT-1 expression in prostate adenocarcinoma Gleason scored > 7 was lower than prostate adenocarcinoma Gleason score ≤ 7, but could not be analyzed the difference significance with BPH because there was only 1 BPH sample in this research. Conclusion: Zn transporters have the potential to be a prognostic factor of prostate adenocarcinoma.;Background: Decreased levels of Zinc ( Zn ) in the prostate correlates with an increase in the grade Gleason score of prostate adenocarcinoma and decrease in Caspase-3 as apoptosis executor. Zn transporter expression levels in prostate tumor cells associates with the level of malignancy. The ZnT-1 is Zn exporter, while the ZIP-1 is Zn importer of prostate cells. ZIP-1 expression drops on adenocarcinoma prostate. Correlation ZnT-1, ZIP-1 and Caspase-3 allegedly influential in prostate carcinogenesis and potential to be prognostic factor of prostate adenocarcinoma.

Design: A cross-sectional retrospective study analytic . Methodology: The sample is 31 paraffin blocks were grouped into BPH, prostate adenocarcinoma Gleason scored ≤ 7 and prostate adenocarcinoma Gleason scored > 7. Samples are analyzed expression of ZnT-1 by immunohistochemical staining. ZIP-1 and Caspase-3 expression is secondary data of Septiawan et al’s immunohistochemical staining. Results: ZnT-1 expression correlated with gleason score. ZIP-1 correlated with the activation of Caspase-3 in prostate adenocarcinoma and prostate adenocarcinoma Gleason score ≤ 7. ZnT-1 correlated with ZIP-1 in prostate adenocarcinoma Gleason score > 7. ZnT-1 expression in prostate adenocarcinoma Gleason scored > 7 was lower than prostate adenocarcinoma Gleason score ≤ 7, but could not be analyzed the difference significance with BPH because there was only 1 BPH sample in this research. Conclusion: Zn transporters have the potential to be a prognostic factor of prostate adenocarcinoma., Background: Decreased levels of Zinc ( Zn ) in the prostate correlates with an increase in the grade Gleason score of prostate adenocarcinoma and decrease in Caspase-3 as apoptosis executor. Zn transporter expression levels in prostate tumor cells associates with the level of malignancy. The ZnT-1 is Zn exporter, while the ZIP-1 is Zn importer of prostate cells. ZIP-1 expression drops on adenocarcinoma prostate. Correlation ZnT-1, ZIP-1 and Caspase-3 allegedly influential in prostate carcinogenesis and potential to be prognostic factor of prostate adenocarcinoma.

Jakarta: Fakultas Kedokteran Universitas Indonesia, 2014

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Loeki Enggar Fitri

Expression of inducible nitric oxide synthase, caspase-3 and production of reactive oxygen intermediate on endothelial cells culture (HUVECs) treated with P. falciparum infected erythrocytes and tumour necrosis factor-a

"Cytoadherence eritrosit terinfeksi P.falciparum pada sel endolel adalah faktor utama dalam perkembangan malaria berat. Proses ini diduga melibatkan respon iinun lokal yang distimulasi o!eh Tumour Necrosis Factor-a (TNF-a). Penelitian ini dilakukan untuk mengetahui pengaruh Cytoadherence eritrosit terinfeksi P.falciparum dan pemberian TNF-a dalam mengtnduksi aktivasi sel endotel secara in vitro. Ekspresi inducible Nitric Oxide Synthase (iNOS) dan Caspase-3, serta produksi Reactive Oxygen Intermediate (ROl) digunakan sebagai parameter aktivasi. Suatu penelitian eksperimental laboratorium telah dilakukan untuk melihat aklivasi sel endotel (HUVECs) setelah dipajan dengan TNF-a selama 20 jam atau eritrosit terinfeksi P.falciparum selama 1 jam atau keduanya. Biakan sel endotel normal digunakan sebagai kontrol. Dengan metode imunohistokimia respon imun lokal dari sel endotel ditentukan dengan melihat ekspresi iNOS dan Caspase 3. Nitro Blue Tetrazolium reduction-assay dilakukan untuk melihat produksi ROi secara semikwantitatif pada sel endotel. Ekspresi iNOS hanya ditemiikan pada biakan sel endotel yang dipajan dengan eritrosit terinfeksi P.falciparum atau eritrosit terinfeksi P.falciparum bersama TNF-a. Ekspresi Caspase 3 terlihat tipis pada beberapa sel pada biakan endotel normal. Ekspresi ini meningkat secara signifikan pada biakan sel endotel yang dipajan eritrosit terinfeksi P.falciparum bersama TNF-a (p=0,000). Sel endotel normal yang diinduksi dengan suaiu induktor non spesifik (PMA) mengeluarkan ROl dalam kadar yang sangat rendah. Pemberian eritrosit terinfeksi P.falciparum saja atau eritrosit terinfeksi P.falciparum bersama TNF-a menyebabkan sel endotel mengeluarkan ROf dalam kadar medium sampai finggi. Pajanan eritrosit terinfeksi P.falciparum dan TNF-a pada sel endotel dapat menginduksi respon iinun lokal yang ditandai dengan peningkatan inducible nitric oxide synthase dan pelepasan radikal bebas sehingga menyebabkan kerusakan sel. (Med J Indones 2006; 15:151-6)

Cytoadherence of P. falciparum infected erythrocytes on endothelial cells is a key factor in development of severe malaria. This process may associated with the activation of local immune that was enhanced by Tumour Necrosis Factor-a (TNF-a). This study was conducted to see the influence of P.falciparum infected erythrocytes Cytoadherence and TNF-a treatment in inducing endolhelial cells activation in vitro. Inducible Nitric Oxide Synthase (iNOS) and Caspase-3 expression, also Reactive Oxygen Intermediate (ROl) production were used as parameters. An Experimental laboratory study had been done to obsen'e endothelial cells activation (HUVECs) after treatment with TNF-a for 20 hours or P.falciparum infected erythrocytes for I hour or both of them. Normal endolhelial cells culture had been used as a control. Using immunocytochemistry local immune activation of endothelial cells was determined by iNOS and Caspase-3 expression. Nitro Blue Tetrazolium reduction-assay was conducted to see the ROl production semi quantitatively. Inducible Nitric Oxide Synthase expression only found on endothelial cells culture treated with P.falciparum infected erythrocytes or both P.falciparltm infected erythrocytes and TNF-a. Caspase-3 expression found slightly on normal endothelial cells culture. This expression increased significantly on endothelial cells culture treated with both P.falciparum infected erythrocytes and TNF-a (p=0,000). The normal endothelial cells release low level of ROi in the presence of non-specific trigger, PMA. In the presence of P.falciparum infected erythrocytes or TNF-a or both of them, some celts showed medium to high levels of ROI. Cytoadherence of P. falciparum infected erythrocytes and 77VF a treatment on endothelial cells can induce activation of local immune marked by increase inducible nitric oxide synthase and release of free radicals that cause cell damage. (Med J'Indones 2006; 15:151-6)"

[place of publication not identified]: Medical Journal of Indonesia, 2006

MJIN-15-3-JulySept2006-151

Artikel Jurnal Universitas Indonesia Library

Aditya D. Septiawan

Analysis expression of ZIP1 and caspase-3 protein in adenocarsinoma of the prostate

"Objective: Carcinogenesis of adenocarcinoma of the prostate occurs due to dysregulation of zinc level within the cells. Intracellular zinc molecules influx is regulated by a transporter protein ZIP1, whose non-presence is predicted to inhibit apoptosis, thus leads to the development of prostate adenocarcinoma. Methods: This study was aimed to analyze the correlation of ZIP1 and Caspase-3 expression in prostate adenocarcinoma on its grading as represented by Gleason Score. This was a cross-sectional, retrospective analytical study on 31 formalin-fixed, paraffin-embedded tissue that meets inclusion criteria. The specimen was stained using the immune-histochemistry technique for ZIP1 and Caspase-3. Protein expression of each case was counted using ImageJ analysis. Gleason score was acquired as secondary data from the cases?s reports. The correlation of their expression with respect to Gleason score was analyzed with Pearson?s correlation using SPSS 11.5. Results: Mean expression level of ZIP1 and Caspase-3 in prostate adenocarcinoma were 35% and 33%, respectively. There was a significantly positive correlation between ZIP1 and Caspase-3 expression (r = 0.379; p = 0.018). However, their correlation was stronger in intermediate-grade group (r = 0.73; p = 0.01) and the correlation was much weaker in high-grade group (r = 0.04; p = 0.48). Conclusions: There was a positive correlation between ZIP1 and Caspase-3 expression in prostate adenocarcinoma.;"

Jakarta: Fakultas Kedokteran Universitas Indonesia, 2016

J-Pdf

Artikel Jurnal Universitas Indonesia Library

Microbiota composition, HSP70 and caspase-3 expression as marker for colorectal cancer patients in aceh, Indonesia / Fauzi Yusuf, Syafruddin Ilyas, Harun AR. Damanik, Fatchiyah

"Tujuan: menganalisis hubungan komposisi mikrobiota terhadap ekspresi HSP70 dan Caspase-3 pada jaringan kolon pasien yang menjalani kolonoskopi dalam upaya pengembangan kandidat deteksi dini untuk

pasien kanker kolorektal di Indonesia. Metode: penelitian potong lintang dilakukan pada 32 responden yang

menjalani pemeriksaan kolonoskopi. Selanjutnya diketahui bahwa 16 orang adalah penderita kanker kolorektal,

sementara 16 lainnya bukan kanker kolorektal (yaitu kolitis dan hemorrhoid interna). Komposisi mikrobiota

pada sampel feses diperiksa dengan menggunakan 16S rRNA Denaturing Gradient Gel Electrophoresis

(DDGE) sedangkan pemeriksaan immunohistokimia untuk menilai ekspresi HSP70 dan Caspase-3 diperiksa

dengan pewarnaan Haematoxylin-Eosin(HE) untuk mengetahui perubahan morfologis pada jaringan kolon.

Hasil: analisis dengan PCR-DGGE menunjukkan perbedaan komposisi mikrobiota yang terdapat pada pasien

kanker kolorektal dan bukan penderita kanker kolorektal. Semua pasien dengan kanker kolorektal menunjukkan

hilangnya pita dominan pada kelompok Bifidobacterium. Pengamatan histologi yang dihitung berdasarkan

uji Inter Class Corelation (ICC) didapati skor yang cukup tinggi (5,2-9,2) pada pasien kanker dan skor yang

lebih rendah (1,7-2,4) pada pasien bukan kanker kolorektal. Ekspresi HSP70 mengalami peningkatan secara

signifikan pada pasien kanker kolorektal dengan persentase tertinggi yaitu 84%, sebaliknya, ekspresi Caspase-3

mengalami penurunan dengan persentase tertinggi hanya 21%. Hasil analisis statistik menunjukkan bahwa

kejadian kanker kolorektal berhubungan dengan ekspresi HSP70 (p<0,001) dan berhubungan dengan ekspresi

Caspase-3 (p<0,001). Kesimpulan: penelitian ini mengindikasikan bahwa Bifidobacterium menjadi indikator

penting terhadap pasien kanker kolorektal yang ditunjukkan pada gambaran pita yang menghilang, sedangkan

ekspresi HSP 70 mengalami peningkatan dan ekspresi Caspase 3 terjadi penurunan yang signifikan.

Aim: to investigate the relationship between microbiota composition with HSP70 and Caspase-3 expressions

in colon tissue as an initial study to develop the candidate for early detection of colorectal cancer for Indonesian

patients. Methods: this is a cross-sectional study on 32 patients undergoing colonoscopy; 16 patients of colorectal

cancer (CRC) while the other 16 patients are not (colitis and internal hemorrhoid). The composition of microbiota

in stool samples was examined using 16S rRNA Denaturing Gradient Gel Electrophoresis (DDGE) while expression

of HSP70 was examined by immunohistochemistry and Caspase-3 by using Haematoxylin-Eosin(HE) staining to determine the morphological changes in colon tissue. Results: analysis of PCR-DDGE shows a different composition

of microbiota between patients with CRC and non-CRC. All CRC patients showed disappearance of dominant band

from Bifidobacterium groups. Histological observation based on Inter Class Correlation (ICC) test from all slide

showed a high scores (5.2-9.2) in CRC patients and low scores (1.7-2.4) in non-CRC patients. HSP70 expression

was increased significantly in CRC patients with the highest percentage of 84%, while expression of caspase-3

decreased with the highest percentage of 21%. Statistical analysis showed that the incidence of colorectal cancer

was associated with the expression of HSP 70 (p<0.001), and Caspase 3 (p<0.001). Conclusion: bifidobacterium is

an important indicator for colorectal cancer patients that show disappearance of dominant band, while expression

of HSP70 increased and the Caspase-3 expression decreased significantly."

Syiah Kuala University. Faculty of Medicine ; North Sumatera University. Mathemathical and Natural Science Faculty ; North Sumatera University. Faculty of Medicine ; Brawijaya University. Mathemathical and Natural Science Faculty, 2016

610 IJIM 48:4 (2016)

Artikel Jurnal Universitas Indonesia Library

Aditya D. Septiawan

Analysis expression of ZIP1 and caspase-3 protein in adenocarsinoma of the prostate / Aditya D Septiawan, Ria Kodariah, Meilania Saraswati

"Objective: Carcinogenesis of adenocarcinoma of the prostate occurs due to dysregulation of zinc level within the cells.

Intracellular zinc molecules influx is regulated by a transporter protein ZIP1, whose non-presence is predicted to inhibit

apoptosis, thus leads to the development of prostate adenocarcinoma. Methods: This study was aimed to analyze the

correlation of ZIP1 and Caspase-3 expression in prostate adenocarcinoma on its grading as represented by Gleason

Score. This was a cross-sectional, retrospective analytical study on 31 formalin-fixed, paraffin-embedded tissue that

meets inclusion criteria. The specimen was stained using the immune-histochemistry technique for ZIP1 and Caspase-3.

Protein expression of each case was counted using ImageJ analysis. Gleason score was acquired as secondary data from

the cases’s reports. The correlation of their expression with respect to Gleason score was analyzed with Pearson’s

correlation using SPSS 11.5. Results: Mean expression level of ZIP1 and Caspase-3 in prostate adenocarcinoma were

35% and 33%, respectively. There was a significantly positive correlation between ZIP1 and Caspase-3 expression

(r = 0.379; p = 0.018). However, their correlation was stronger in intermediate-grade group (r = 0.73; p = 0.01) and the

correlation was much weaker in high-grade group (r = 0.04; p = 0.48). Conclusions: There was a positive correlation

between ZIP1 and Caspase-3 expression in prostate adenocarcinoma."

Fakultas Kedokteran Universitas Indonesia, 2016

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Artikel Jurnal Universitas Indonesia Library

Elvira Yunita

Efek in vitro andrografolida terhadap apoptosis jalur intrinsik pada sel punca kanker payudara manusia yang dipaparkan rotenon: tinjauan ekspresi caspase-9, caspase-3 dan survivin = In vitro effect of andrographolide on the intrinsic pathway of apoptosis in rotenone induced human breast cancer stem cells focus on caspase 9 caspase 3 and survivin / Elvira Yunita

"ABSTRAK

Nama : Elvira YunitaProgram Studi : Program Magister Ilmu BiomedikJudul : Efek In Vitro Andrografolida terhadap Apoptosis Jalur Intrinsik pada Sel Punca Kanker Payudara Manusia yang Dipaparkan Rotenon: Tinjauan Ekspresi Caspase-9, Caspase-3 dan Survivin. Latar belakang: Andrografolida ANDRO merupakan senyawa bioaktif utama yang berasal dari sambiloto, Andrographis paniculata. Penelitian in silico yang telah dilakukan menunjukan ANDRO mempengaruhi apoptosis intrinsik dengan berinteraksi dengan survivin, caspase-9 dan caspase-3. Penelitian lain menunjukkan bahwa Breast Cancer Stem Cell BCSC memiliki survival rate yang lebih tinggi setelah dipaparkan dengan rotenon jika dibandingkan dengan non-BCSC. Oleh karena itu penelitian ini bertujuan untuk menganalisis apoptosis jalur intrinsik pada BCSC yang telah dipaparkan rotenon dan ANDRO.Metode: BCSC dipaparkan dengan 50 ?M rotenon selama 6 jam dan kemudian sel dipaparkan dengan ANDRO 0.075 mM, 0.15 mM, 0.3 mM dan 0.6 mM selama 24 jam. RNA sel diisolasi dan diikuti dengan pengukuran ekspresi mRNA caspase-9 dan 3 dengan qRT-PCR. Protein sel akan digunakan untuk pengukuran ekspresi protein survivin total dan survivin terfosforilasi. Selain itu, Apoptosis sel juga diukur dengan flowcytomety.Hasil: Perlakuan dengan rotenon dan ANDRO dapat meningkatkan ekspresi mRNA caspase-9 dan caspase-3 yang juga disertai dengan penurunan viabilitas. Paparan ANDRO dengan konsentrasi rendah hingga 0.015 mM pada BCSC yang telah diberikan rotenon dapat menurunkan ekspresi mRNA survivin. Dosis yang lebih tinggi dapat meningkatkan ekspresi mRNA survivin. Ekspresi protein survivin mengalami peningkatan yang disertai dengan penurunan rasio survivin yang teraktivasi. Selain itu, persentase apoptosis setelah paparan rotenon dan ANDRO hingga 0.3 mM ANDRO juga mengalami peningkatan seiring dengan bertambahnya konsentrasi ANDRO yang dipaparkan dose dependent manner .Kesimpulan: Paparan rotenon dan ANDRO dapat menginduksi kematian sel melalui apoptosis jalur intrinsik pada BCSC. Selain itu, ANDRO juga diusulkan sebagai senyawa yang potensial menjadi agen terapi pendamping bagi pasien-pasien yang menjalani kemoterapi maupun radioterapi.

ABSTRACT

ABSTRACT Name Elvira YunitaStudy Program Biomedical ScienceTitle In Vitro Effect of Andrographolide on the intrinsic pathway of apoptosis in rotenone induced human breast cancer stem cells Focus on caspase 9, caspase 3 and survivin expression Background Andrographolide ANDRO is an active compound of Andrographis paniculata, has been suggested to have an anti cancer property. Our in silico study has shown that ANDRO could interact with survivin, caspase 9 and caspase 3 which influence the intrinsic apoptotic pathway. We have also demonstrated that human breast cancer stem cells BCSCs could survive better than their counterpart non BCSCs after rotenone treatment. In this study, we aimed to investigate whether andrographolide could induced apoptotic of rotenone induced BCSCs.Method Human BCSCs ALDH cells were first induced by 50 M rotenone for 6 hours and treated with 0.075 mM, 0.15 mM, 0.3 mM dan 0.6 mM of ANDRO for 24 hours. Total RNA from the cells was isolated, followed by determination of survivin, caspase 9 and caspase 3 m RNA expression level using Real Time RT PCR technique. Protein from the cells used to examine expression of survivin and phosphorylated survivin. Besides, examining of apoptotic cell also measured by flowcytometry.Result Treatment of rotenone and ANDRO drastically increase caspase 9 and caspase 3 expression, leading to the decrease of cell viability. Low concentration of ANDRO treatment until 0.15 mM could supress survivin expression, but gradually increased higher than the control in line with the increasing of ANDRO concentration. Survivin protein expression was increased following by decreasing ratio of activated survivin. Besides, percentage of apoptosis after rotenone and ANDRO treatment until 0.3 mM of ANDRO treatment also increased in line with increasing ANDRO concentration dose dependent manner .Conclusion Rotenone and ANDRO treatment could induced apoptotic intrinsic in BCSCs. Thus, we propose that andrographolide as potential compound that could be used as a novel co chemoradiation therapy and radiotherapy targeted to BCSCs."

2017

T55641

UI - Tesis Membership Universitas Indonesia Library

Ayu Suraduhita

Efek andrografolida pada apoptosis intrinsik pada sel punca kanker payudara (CD24-/CD44+) yang telah mengalami penurunan sensitivitas terhadap pemberian doksorubisin berulang = Effect of andrographolide on the intrinsic apoptosis in breast cancer stem cells (CD24-/CD44+) after decreased sensitivity to repeated doxorubicin administration

"Latar Belakang: Penggunaan doksorubisin untuk terapi kanker masih menjadi pilihan utama karena terbukti poten.Namun, terjadinya resistensi pada sel kanker terhadap kemoterapi merupakan salah satu penghambat keberhasilan dari pengobatan ini. Sel punca kanker payudara berperan pada terjadinya resistensi terapi yang ditandai dengan adanya peningkatan ekspresi survivin. Survivin adalah protein bifungsional yang dapat menekan apoptosis dan mengatur pembelahan sel. Penelitian terbaru menyarankan untuk mengkombinasikan terapi kanker konvensional dengan senyawa aktif bahan alam untuk mencegah resistensi sel kanker terhadap kemoterapi seperti andrografolida. Oleh karena itu senyawa andrografolida diharapkan dapat digunakan sebagai kemosensitizer terhadap doksorubisin. penelitian ini bertujuan untuk menganalisis peran andrografolida dalam mengatasi resistensi dan meningkatkan efektivitas doksorubisin pada sel punca kanker payudara melalui penekanan aktivitas survivin pada jalur apoptosis intrinsik.

Metode: Sel punca kanker payudara (CD24-/CD44+) diberikan doksorubisin 0,1μM. Setelah 14 hari perlakuan maka dilakukan kombinasi doksorubisin 0,1μM dan andrografolida 0,285 mM hingga hari ke-22. Setiap 2 hari, sel dipanen dan dihitung dengan metode eksklusi trypan blue. Analisis ekspresi mRNA Caspase-9, Caspase-3, dan Survivin dilakukan dengan qRT-PCR, sedangkan uji apoptosis dilakukan dengan metode flow cytometry.

Hasil: Pemberian doksorubisin tunggal dapat mengurangi viabilitas sel punca kanker payudara (CD24-/CD44+). Setelah 12 hari pemberian, viabilitas sel punca kanker payudara (CD24-/CD44+). dan ekspresi mRNA survivin meningkat, tetapi ekspresi mRNA caspase 9 dan caspase 3 ditekan. Sedangkan kombinasi doksorubisin dan andrografolida dapat menurunkan viabilitas sel punca kanker payudara (CD24- /CD44+) pada hari ke 16, sejalan dengan penurunan ekspresi survivin mRNA dan peningkatan ekspresi mRNA caspase-9 dan caspase-3.

Kesimpulan: Andrografolida dapat berfungsi sebagai kemosensitizer untuk meningkatkan apoptosis intrinsik pada sel punca kanker payudara yang telah diberikan doksorubisin berulang.

Background: Doxorubicin is still the main option for cancer treatment because it has proven to be effective. However, the resistance of cancer cells to chemotherapy is one of the obstacles to the success of this treatment. Breast cancer stem cells play a role in the development of treatment resistance, which is indicated by the increase in survivin expression. Survivin is a bifunctional protein that suppresses apoptosis and regulates cell division. Recent studies had suggested using additional substances to prevent cancer cell resistance to chemotherapy such as andrographolide. Hence, andrographolide compounds are expected to be used as chemosensitizer against the doxorubicin. This study aimed to analyze the role of andrographolide to overcome the resistance and improve the effectiveness of doxorubicin in human BCSC through suppressing survivin activity on the intrinsic apoptosis pathway.
Method: BCSCs (CD24-/CD44+) were treated with 0.1μM doxorubicin. After 14 days of treatment, the cells were treated with a combination of 0.1μM doxorubicin and 0.285 mM andrographolide until the 22nd day. Every 2 days, the cells were harvested and counted by using the trypan blue exclusion method. The analysis of Caspase-9, Caspase-3, and Survivin mRNA expression was performed by using qRT-PCR, while apoptotic assay was done using flow cytometry.
Result: The single treatment of doxorubicin could reduce BCSCs viability. After 12 days of treatment, the survivin mRNA expression was increased following BCSCs viability, but the caspase 9 and caspase 3 mRNA expressions were suppressed. Meanwhile, the combination of doxorubicin and andrographolide could decrease BCSCs viability on the 16th day, in line with the decreased expression of survivin mRNA, there was an improvement of caspase-9 and caspase-3 mRNA expression levels.
Conclusion: Andrographolide could be considered as chemosensitizer to increase intrinsic apoptosis in breast cancer stem cells that given repeated doxorubicin administration."

Depok: Fakultas Kedokteran Universitas Indonesia, 2020

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