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DATE FRUIT EXTRACT (PHOENIX DACTYLIFERA, AJWA) MODULATES NK CELLS AND TNF- ALPHA IN DMBA-INDUCED MAMMARY CANCER SPRAGUE-DAWLEY RATS Elhemeidy, Rehab Mohammed Mustofa; Lyrawati, Diana; Widjajanto, Edi
Journal of Tropical Life Science Vol 8, No 3 (2018)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.08.03.04

Abstract

Cellular immune system is one of anti-tumor target development. NK cells, one of major components of antitumor innate immune responses, are frequently decreased functionally during breast cancer progression, whereas TNF-alpha changes from anti-tumor into pro-tumorigenesis and its level increases with advancing cancer. In this study, we investigated the immunomodulatory effects of Ajwa date fruit extract to determine whether it stops the progression of breast cancer. Using rats induced with single dose 20 mg DMBA subcutaneously, we examined whether administration of Ajwa date fruit (Phoenix dactylifera) extract (at 400, 800 mg/kg body weight/d) post-DMBA induction for 30 days modulate NK cells, TNF-alpha, and development of breast cancer. ELISA, FACS, immunohistochemistry, and histologic observation were employed. Compared to positive control group (DMBA-induced only), we showed that the Ajwa date fruit extract normalize the level of circulatory CD161 NK cells and breast tissue TNF-alpha, cell size and proliferation, and improve overall survival rates. Ajwa date fruit extract may be used to modulate NK cells and TNF-alpha against progression of breast cancer. Further studies are required to develop date fruit extract as anti-cancer in human, whether as single or adjunct therapy to standard management of breast-cancer.
EFEK CATECHINS TEH HIJAU TERHADAP EKSPRESI ROR αDAN C/EBP-α PADA KULTUR PRIMER PREADIPOSIT Khaira H, Faridha; Ratnawati, Retty; Lyrawati, Diana
Jurnal Kedokteran Brawijaya Vol 27, No 3 (2013)
Publisher : Fakultas Kedokteran Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jkb.2013.027.03.2

Abstract

Obesitas  merupakan  faktor  risiko  terjadinya  berbagai  penyakit  yang  dapat  menyebabkan  kematian.  Pada  tingkat  sel, obesitas dicirikan oleh pertambahan ukuran dan jumlah sel adiposit diantaranya melalui proses adipogenesis. T eh hijau merupakan minuman yang dikonsumsi di seluruh  dunia, yang memiliki banyak manfaat  terhadap kesehatan terutama disebabkan  oleh  konsentrasi  polifenol  catechins  yang  memiliki  efek  anti-obesitas. Penelitian  ini  bertujuan  untuk membuktikan efek isolat  senyawa golongan catechins  teh hijau pada regulator negatif  maupun positif pada adipogenesis yaitu  ROR-?  dan  C/EBP-?.  Penelitian  ini  dilakukan  secara  in  vitro  pada  kultur  primer  preadiposit  dengan  4  kelompok perlakuan:  (1)  kelompok  kontrol,  (2)  dipapar  dengan  catechins  5µM,  (3)  dipapar  dengan  catechins  10  µM  dan  kelompok  (4) dipapar  dengan  catecins  30  µM.  Dua  hari  setelah  sel  mencapai  confluent,  sel  diinduksi  untuk  berdiferensiasi  menjadi adiposit  dengan  IBMX,  DEX  dan  insulin.  Pada  hari  ke  3  diferensiasi,  sel  dipapar  dengan  catechins  selama  24  jam,  setelah  itu sel  dipanen  dan  ekspresi  ROR-?  dan  C/EBP-?  diperiksa  menggunakan  metode  ELISA  dan  immunositokimia.  Isolat  senyawa golongan catechins teh hijau pada dosis 10 dan 30 µM   secara signifikan menurunkan ekspresi C/EBP-? masing-masing sebesar 20,24% dan 26,43%. Pemaparan isolat senyawa golongan catechins teh hijau tidak memiliki efek yang signifikan terhadap  ekspresi  ROR-?  meskipun  ekspresinya  cenderung  meningkat  seiring  dengan  pertambahan  dosis.
Aloe Gel Enhances Angiogenesis in Healing of Diabetic Wound Sargowo, Djanggan; Handaya, Adeodatus Yuda; Widodo, Mohammad Aris; Lyrawati, Diana; Tjokroprawiro, Askandar
The Indonesian Biomedical Journal Vol 3, No 3 (2011)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v3i3.152

Abstract

BACKGROUND: Diabetic micro and macroangiophathy lead to the incident of diabetic foot ulcers characterized by an increased number of circulating endothelial cells (CECs) and decreased function of endothelial progenitor cells (EPCs). This fact is correlated with ischemia and diabetic wound healing failure. Aloe vera gel is known to be able to stimulate vascular endothelial growth factor (VEGF) expression and activity by enhancing nitric oxide (NO) production as a result of nitric oxide synthase (NOS) enzyme activity. Aloe vera is a potential target to enhancing angiogenesis in wound healing.OBJECTIVE: The objective of this study was to explore the major role of Aloe vera gel in wound healing of diabetic ulcers by increasing the level of EPCs, VEGF, and endothelial nitric oxide synthase (eNOS), as well as by reducing the level of CECs involved in angiogenesis process of diabetic ulcers healing.METHODS: The experimental groups was divided into five subgroups consisting of non diabetic wistar rats, diabetic rats without oral administration of aloe gel, and treatment subgroup (diabetic rats) with 30, 60 and 120 mg/day of aloe gel doses for 14 days. All subgroups were wounded and daily observation was done on the wounds areas. Measurement of the number of EPCs (CD34), and CECs (CD45 and CD146) was done by flowcytometry, followed by measurement of VEGF and eNOS expression on dermal tissue by immunohistochemical method on day 0 and day 14 after treatment. The quantitative data were analyzed by One-Way ANOVA and Linear Regression, with a cofidence interval 5% and significance level (p<0.05) using SPSS 16 software to compare the difference and correlation between wound diameters, number of EPCs and CECs as well as the levels of VEGF and eNOS.RESULTS: The results of this study showed that aloe gel oral treatment in diabetic wistar rats was able to accelerate the wound healing process. It was shown by significant reduction of wound diameter (0.27±0.02); the increased number of CECs (0.42±0.57), respectively (p<0.05). On the other hand, the wound diameter and eNOS indicators showed significant differences at the dose of 60 mg, while the number of EPCs and CECs and the level of VEGF showed significantly different results at a dose of 120 mg. Aloe gel oral therapy showed a positive indication of wound healing acceleration at the optimum dose range 60-120 mg a day.CONCLUSIONS: Aloe gel is potential to be a herbal therapy candidate for diabetic wound healing through enhancing EPCs homing, decreasing the CECs number, and stimulating the increase of VEGF and eNOS levels,hence proving to be a dominant factor in the angiogenesis process.KEYWORDS: aloe gel, diabetes, wound healing, angiogenesis
NANOSECOND PULSED ELECTRIC FIELDS (NSPEFS) INDUCE SOCS1 AND SOCS3 BUT NOT SOCS2 GENE EXPRESSIONS IN HELA S3 CELLS Rohmah, Martina Kurnia; Lyrawati, Diana; Yano, Kenichi
Jurnal Kedokteran Brawijaya Vol 28, No 3 (2015)
Publisher : Fakultas Kedokteran Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jkb.2015.028.03.1

Abstract

Nanosecond Pulsed Electric Fields (nsPEFs) is one of bioelectric technologies applied widely in a number of sciences. nsPEFs cause some biological responses and known to play a role as a novel cancer therapy. However, the nsPEFs molecular mechanisms have not been fully elucidated. This study determines the effects of nsPEFs in socs (Suppressor of Cytokine Signaling) genes which are target genes of JAK/STAT signaling pathway. Through a negative feedback mechanism, SOCS proteins can suppress both cytokine signal transduction and overgrowth factor, so the cell growth is controlled. In cervix cancer, the presence of E6 and E7 HPV's oncoprotein is associated with methylation and inactivation of socs1 and socs3 genes. This mechanism is related to the increase of STAT expression and cancer prognostic. In this research, nsPEFs as much as 20 kV/cm for 80 ns was exposed over HeLa S3 cells in 4 mm cuvette. Socs1, socs2 and socs3 gene expressions were analyzed using real time PCR SYBR green and reverse transcription PCR (RT-PCR). This study shows that at 20 and 30 shots, nsPEFs significantly increase socs1 and socs3 but not socs2 gene expression. Effect of nsPEFs on socs1 and socs3 gene expression pattern is influenced by duration of post exposure incubation and each cell activity on internal cell condition. This research provides a new cancer therapy target for nsPEFs.Keywords: Bioelectric, gene expression, nsPEFs, shot, socs gene
Cloning and expression of pab gene of M. tuberculosis isolated from pulmonary TB patient in E.coli DH5α M. Raras, Tri Y.; Lyrawati, Diana
Medical Journal of Indonesia Vol 20, No 4 (2011): November
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (4108.055 KB) | DOI: 10.13181/mji.v20i4.458

Abstract

Background: Mycobacterium tuberculosis antigen38 is a potent serodiagnostic agent containing two M. tuberculosisspecific B-cell epitopes. The high price of imported diagnostic agents hinders realization of fast clinical TB diagnosis in developing countries. Therefore, we produced recombinant antigen38 (recAg38M) from M. tuberculosis local strain, which might be used to produce economical tuberculosis serodiagnostic kit.Methods: Pab gene that was isolated from pulmonary TB patient in Malang was cloned into a plasmid vector (pGEMTeasy) to construct pMB38. The E.coli DH5α clone carrying pMb38 was selected on X-gal medium. The expression of pab was mediated using pPRoExHTc under the control of Trc promoter and E.coli DH5α as host.Results: Alignment of the pab sequence from the white E.coli DH5α clones with that of M. tuberculosis H37Rv showed 98% homology. The recombinant protein in which the signal peptide has been deleted to prevent the protein being secreted into medium was found in the cytoplasm.Conclusion: pab gene of M. tuberculosis isolated from a TB patient could be expressed in heterologous system in E.coliDH5α. (Med J Indones 2011; 20:247-54)Keywords: Mycobacterium tuberculosis, Pab gene expression, recombinant antigen38
EKSPRESI CYTOSOLIC ASPARTATE-SPECIFIC CYSTEINE PROTEASE-3 (CASPASE-3) PADA JARINGAN HATI RATTUS NORVEGICUS (WISTAR) SETELAH PEMBERIAN SUBKRONIK AFLATOKSIN B1 (AFB1) Maritha, Indah Dina; Supranowo, Supranowo; Lyrawati, Diana
Jurnal Kedokteran Brawijaya Vol 22, No 3 (2006)
Publisher : Fakultas Kedokteran Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jkb.2006.022.03.4

Abstract

Aflatoksin B1(AFB1) is one of the toxic agents produced by Aspergillus flavus that frequently contaminates foods not properly stored. AFB1 undergoes biotransformation which may result in the production of reactive oxygenspecies (ROS) hazardous to liver cells. Following a cascade of oxidative reaction, ROS will cause the mitochondria torelease cytochrome c which subsequently activates caspase-3, leading to apoptosis. In this present study we evaluated the effect of aflatoxin B1 on the expression of caspase-3 in the liver. AFB1 was administered per oral, at different dosage and length of exposure, subchronically. This study was carried out as a factorial designed experiment with  two factors. The first was dosage factor i.e 0, 10, 15 and 20 µg (0; 0,05; 0,075; 0,1 µg/g BW) and the second was exposure time factor i.e. 12, 16 and 20 weeks. Rattus norvegicus strain Wistar aged approximately eight weeks old and weighed 180-200 g were used as the experimental animals. The expression of  caspase-3 was examined by using immunohistochemistry. The results showed that the expression of caspase-3 increased significantly (p = 0,000) with the escalation of AFB1 in dosage and/or exposure time (p = 0,001). In the interaction between dose and exposure time of AFB1an increase in the expression of caspase-3 was also observed (p = 0,000). Interestingly, these studies also revealed thatin the liver tissues there was a limitation in the expression of caspase-3, where the raising of further AFB1 dosage and length of exposure were not followed by further increase of the caspase-3.
EFEK PEMBERIAN ASAM ALFA LIPOAT TERHADAP KADAR MDA DAN GAMBARAN HISTOLOGI PADA HATI TIKUS WISTAR JANTAN DENGAN DIABETES MELITUS TIPE 1 Fitria, Nur Lailatul; Lyrawati, Diana; Handaru, Mudjiwijono
Jurnal Kedokteran Brawijaya Vol 28, No 3 (2015)
Publisher : Fakultas Kedokteran Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jkb.2015.028.03.2

Abstract

Diabetes melitus dan komplikasinya disebabkan oleh hiperglikemia yang menyebabkan stres oksidatif melalui produksi spesies oksigen reaktif (ROS). Asam Alfa Lipoat (ALA) terbukti dalam mengatasi stres oksidatif yang menyebabkan komplikasi vaskular pada diabetes. Penelitian ini bertujuan untuk mengetahui efek ALA terhadap kadar malondialdehid (MDA) dan gambaran histologi hati tikus diabetes mellitus tipe 1. Rancangan penelitian ini menggunakan true experimental design dan posttest  only  control  group.  Tiga puluh ekor  tikus jantan (Rattus norvegicus galur wistar)  dibagi menjadi  lima  kelompok yaitu:  kelompok  tikus  normal,  tikus diabetes, tikus  diabetes  dengan  ALA  dosis 80mg/kgBB/hari, tikus  diabetes  dengan  ALA  dosis  200mg/kgBB/hari, dan tikus diabetes dengan ALA dosis 500mg/kg/hari. ALA diberikan sehari sekali secara oral. Induksi diabetes pada tikus dilakukan  secara intraperitonial  dosis  tunggal  streptozotocin  60mg/kgBB. Kadar  MDA  pada hati  diukur dengan metode spektrofotometri dan histologi hati diuji  setelah diwarnai dengan hematoksilin dan eosin. Pada penelitian ini, ALA dosis 80 dan 200mg/kgBB/hari tidak memberikan pengaruh terhadap kadar MDA, namun terjadi perbaikan gambaran histologi, warna hati, dan penurunan perlemakan pada hepatosit dibandingkan tikus diabetes. Pada dosis ALA 500mg/kgBB/hari menunjukkan warna hati lebih pucat, peningkatan perlemakan pada hepatosit, dan tingkat kematian yang tinggi. Kesimpulan  dari  penelitian  ini, pemberian  ALA selama 4 minggu tidak berpengaruh terhadap kadar MDA, namun memperbaiki gambaran histologi hati tikus.ata Kunci: Asam alfa lipoat, diabetes mellitus tipe 1, histologi hati, malondialdehid,perlemakan hepatosit
Catechins of GMB-4 Clone Inhibits Adipogenesis Through PPAR-γ and Adiponectin in Primary Culture of Visceral Preadipocyte of Rattus Norvegicus Wistar Nur, Aswaty; Ratnawati, Retty; Lyrawati, Diana
Research Journal of Life Science Vol 5, No 1 (2018)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (423.852 KB) | DOI: 10.21776/ub.rjls.2018.005.01.6

Abstract

Catechins of green tea (Camelia sinensis) GMB4 clone may serve as a potential therapeutic antiobesity agent, probably through its effects on preadipocytes. Thus, to evaluate such antiobesity effects, we performed series of in vitro experiments using primary cultures of visceral preadipocytes from Rattus norvegicus strain Wistar. Quick Cell Proliferation assay, Oil Red-O staining, ELISA and immunocytochemistry were used to determine the effects of 25 µM, 50 µM, 75 µM, and 100 µM catechins on primary culture of preadipocytes, particularly on cell viability and differentiation as well as on expression of relevant obesity genes i. e. PPARγ and adiponectin levels. The results showed that there were no significant differences on preadipocytes viability among control and catechins treatments except in cells treated with 50 µM catechins (means±SD=128±2.47) which resulted 28% higher viability than control (p= 0.037). Catechins inhibited preadipocytes differentiation into adipocytes, at 100 µM up to 78% lower than control. The level of PPARy apparently was reduced by catechins, but statistically significant only at 75 μM (p= 0.029). In contrast, the adiponectin level on preadipocytes increased by catechins at 75 µM and 100 µM (0.786±0.126 and 0.673 ±0.319 ng/ml; control: 0.077±0.017, p ≤0.01). In conclusions, our data revealed that desired antiobesity effects of catechins of green tea GMB4 clone on visceral preadipocytes were concentration-dependent; at dosage 50 µM catechins enhanced cell viability; at more than 75 µM inhibited differentiation of preadipocytes and was associated with lower PPARy and higher adiponectin levels.
ARTERIOGENESIS DAN ANGIOGENESIS PADA STROKE HEMORAGIK:MEMPERTAJAM KONSEP UNTUK MEMPEROLEH MANFAAT TERBAIK NEOVASKULARISASI * Lyrawati, Diana
Jurnal Kedokteran Brawijaya Vol 24, No 1 (2008)
Publisher : Fakultas Kedokteran Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jkb.2008.024.01.6

Abstract

Arteriogenesis and angiogenesis contribute both to  the damage and repair in hemorrhagic stroke. Understanding and unraveling the fundamental and mechanisms of these processes should facilitate the development of treatment strategies to improve outcome after stroke. This article describes briefly the basic principles of arteriogenesis and angiogenesis completed with some key molecular players, followed by aperspective of the author on how to use those neovascularization to benefit repair and recovery processes in post-hemorrhagic stroke. Key Words : arteriogenesis, angiogenesis, hemorrhagic, stroke
Efek Ekstrak Alga Coklat (Sargassum sp.) sebagai Supresor Protein Bcl-2 in vitro pada Sel HeLa Putri, Neshya Ruriana; Lyrawati, Diana; Indra, M. Rasjad
Majalah Kesehatan FKUB Vol 1, No 4 (2014)
Publisher : Faculty of Medicine Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (487.995 KB)

Abstract

Prevalensi kanker rahim di Indonesia masih tinggi. Protein Bcl-2 salah satu protein yang menghambat apoptosis melalui jalur mitokondria. Pada kanker serviks terjadi mutasi p53 sehingga Bcl-2 mengalami ekspresi berlebihan.Terapi kanker serviks menggunakan bahan kimia dapat menimbulkan banyak efek samping. Senyawa anti kanker dari bahan alam yang  sedikit menimbulkan efek samping diharapkan mampu mempengaruhi ekspresi protein Bcl-2. Pada semua jenis Sargassum sp. mengandung fukosantin dengan kadar tinggi. Fukosantin dapat menurunkan ekspresi protein Bcl-2 pada sel kanker. Tujuan penelitian ini adalah membuktikan bahwa pemberian ekstrak alga coklat (Sargassum sp.) dapat menghambat ekspresi Bcl-2 pada kanker serviks (sel HeLa). Penelitian ini merupakan penelitian eksperimental laboratorik dengan rancangan true experimental. Sampel yang digunakan adalah sel HeLa yang terbagi atas 1 kontrol dan 3 perlakuan. Kelompok I adalah kontrol sel HeLa. Kelompok II-IV diberi perlakuan ekstrak alga coklat dengan kadar yang berbeda-beda (31,25 µg/ml, 62,5 µg/ml, dan 125 µg/ml). Semua kelompok diinkubasi selama 24 jam. Parameter yang diukur adalah ekspresi Bcl-2 pada sel HeLa yang dideteksi secara imunohistokimia. Hasil analisis statistika menunjukkan bahwa ekstrak alga coklat menurunkan eskspresi Bcl-2 sel HeLa secara signifikan (ANOVA, p = 0,000; R² = 0,947). Penurunan ekspresi Bcl-2 mulai terlihat sejak pemberian ekstrak Sargassum dengan kadar 31,25 µg/ml. Pemberian ekstrak alga coklat kadar 31,25 µg/ml, 62,5 µg/ml, dan 125 µg/ml menurunkan ekspresi Bcl-2 sebesar 21 %, 44 %, dan 68 %. Kesimpulan penelitian ini adalah ekstrak alga coklat (Sargassum sp.) mampu menurunkan ekspresi Bcl-2 sel HeLa dengan kadar efektif terendah 31,25 µg/ml. Kata kunci: Bcl-2, Ekstrak alga coklat (Sargassum sp), Sel HeLa.