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All Journal Journal of Tropical Life Science : International Journal of Theoretical, Experimental, and Applied Life Sciences Jurnal Kedokteran Brawijaya Medical Journal of Indonesia Research Journal of Life Science Program Kreativitas Mahasiswa - Penelitian INDONESIAN JOURNAL OF PHARMACY MNJ (Malang Neurology Journal) Majalah Kesehatan FKUB
Diana Lyrawati
Laboratorium Farmasi Fakultas Kedokteran Universitas Brawijaya Malang
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science Medicine & Pharmacology Neuroscience Public Health

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Role of Antibody Anti-AGE in the Expression of Nephrin and RAGE on Primary Glomerulus Cell Exposed with AGE Salam, Rudy; Lyrawati, Diana; Samsu, Nur
Journal of Tropical Life Science Vol 7, No 2 (2017)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.07.02.03

Abstract

Upregulation of nephrin expression occurs at the early stage of nephropathy and decrease in the period 6 months on hyperglycaemic condition. Nephrin is associated with the initial stage of the loss of the permeability barrier in diabetic nephropathy. Interaction AGE-RAGE increases angiotensin II on Renin Angiotensin-Aldosterone System (RAAS) and activation of protein kinase c (PKC) which induce alterations in nephrin mRNA expression. Alterations of nephrin expression induces transformation of slit membrane structure and the permeability changes at the glomerular filtration barrier. Anti-AGE vaccination once may cause the changes of nephrin and RAGE expression and can prevent progression of diabetic nephropathy. This study used primary glomerulus cell culture obtained from renal of Wistar mice aged 3 months, weighting 200-300 grams that consist of negative control group that exposed to BSA 100 µg/ml, positive control group that exposed to AGE-BSA 100 µg/ml, treatment group 1 that exposed to polyclonal anti-AGE 5 µg/ml and AGE-BSA 100 µg/ml and treatment group 2 that exposed to monoclonal antibody anti-CML 5 µg/ml and AGE-BSA 100 µg/ml. Paired t-test with a 0.05 level of confidence results showed that there were significant differences in level of RAGE expression among experimental groups with control groups. Administration of polyclonal antibody decreased RAGE expression among negative control (p=0.188). but not in positive control (p=0.000). In contrast to monoclonal anti-AGE antibody, RAGE expression did not differ significantly compared to negative control but significant than positive control. Administration of monoclonal anti-AGE antibody inhibited increasing of nephrin expression compared to negative and positive control (p=0.73; 0.125). In conclusion, this study suggested that administration of polyclonal and monoclonal anti-AGE antibody could inhibit increasing of RAGE and nephrin expression in glomerulus primary culture that exposed to AGE which is expected to prevent the progression of diabetic nephropaty.
ALPHA-PINENE ATTENUATES MICROGLIAL NF-ΚB ACTIVATION AND INOS EXPRESSION IN GP120-INDUCED NEUROINFLAMMATION Rahayu, Masruroh; Widodo, M Aris; Lyrawati, Diana; Widjadjanto, Edi
MNJ (Malang Neurology Journal) Vol. 7 No. 1 (2021): January
Publisher : PERDOSSI (Perhimpunan Dokter Spesialis Saraf Indonesia Cabang Malang) - Indonesian Neurological Association Branch of Malang cooperated with Neurology Residency Program, Faculty of Medicine Brawijaya University, Malang, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.mnj.2021.007.01.16

Abstract

Background: Neuro-inflammation plays a role in the pathogenesis of HIV-associated dementia (HIV). Activation of microglia is essential for triggering inflammatory-mediated neurotoxicity. HIV-1 120 kDa envelope glycoprotein (gp120) induces microglial NF-κB signaling which in turn induce pro-inflammatory and iNOS gene transcription. Continuous or excessive activation of NF-κB signaling lead to persistent production of TNF-α and nitric oxide by microglia and induce neuronal apoptosis. Alpha-pinene is a natural substance found in pine tree and has efficacy on inhibiting NF-κB signaling.Objective: This study was designed as a true experimental study and aimed to investigate the effect of alpha-pinene administration toward inflammatory response represented by the percentage of microglia containing activated NF-κB and iNOS expression.Methods: Neuron-glia primary culture from brain tissue of rat fetus was divided into 5 groups as follows: negative control; positive control (gp120 1nM); treatment I, II, and III (gp120 1 nM + alpha-pinene 0.4 µg/mL, 2 µg/mL, and 10 µg/mL, respectively). Microglial NF-κB and iNOS expression were analyzed using immunohistochemistry method. Neuronal apoptosis was measured by TUNNEL method.Results: Result showed that alpha-pinene administration on gp120-treated neuron-glia at all dosages decrease NF-kB activation, iNOS expression, and apoptotic neuron significantly as compared to the gp120-only treated group (p<0.05). Furthermore, alpha-pinene did not affect NF-kB activation and neuronal apoptosis (p>0.05), but significantly elevate iNOS expression (p<0.05) mainly in dosage I and II.Conclusion: We concluded that alpha-pinene has neuroprotective effect on gp120-treated neuron-glia cells through modulation of NF-kB and iNOS expression thus inhibit neuronal apoptosis.
Advanced Glycation End Products (AGEs) Antibody Protects Against AGEs-induced Apoptosis and NF-ĸB p65 Subunit Overexpression in Rat Glomerular Culture Adianingsih, Oktavia Rahayu; Lyrawati, Diana; Samsu, Nur
Journal of Tropical Life Science Vol 6, No 3 (2016)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.06.03.08

Abstract

Advanced glycation end products (AGEs) have been thought to be a major cause of diabetic nephropathy (DN). The mechanisms underlying the involvement of AGEs antibody in diabetic nephropathy are not fully understood. The present study was designed to investigate the protective effect of AGEs antibody on AGEs-induced glomerular damage. Isolated glomeruli were pre-incubated either with 10 µg/mL polyclonal anti-AGEs antibody (AGE-pAb) or monoclonal anti-Nɜ -carboxymethyl-lysine antibody (CML-mAb) as a model of AGEs antibody to block interaction of AGEs with receptor for AGEs (RAGE) and incubated afterwards either with 100 µg/mL bovine serum albumin (BSA) or AGE-modified bovine serum albumin (AGE-BSA) for 48 h. Annexin V/nephrin doublestaining was performed to determine apoptosis. Using immunofluorescence, we found that administration of AGE-BSA not only significantly increased glomerular cells apoptosis and nuclear factor kappa B (NF-ĸB) p65 expression, but also reduced expression of nephrin, an important structural and signal molecule of podocytes slit diaphragm. Blocking the interaction of AGE-RAGE with AGEs antibody significantly protected glomerular cells from AGEs-induced apoptosis and NF-ĸB p65 overexpression. We found that AGE-pAb conferred superior protective effect compared with CmL-mAb for the same reduction in apoptosis and NF-ĸB p65 expression. In sharp contrast, CmL-mAb led to preserve expression of podocytes nephrin better than AGE-pAb. These results demonstrate that the antibody against AGEs may be beneficial for preventing the glomerular damage in DN.
MONOSODIUM GLUTAMATE EXPOSURE AT EARLY DEVELOPMENTAL STAGE INCREASES APOPTOSIS AND STEREOTYPIC BEHAVIOR RISKS ON ZEBRAFISH (DANIO RERIO) LARVAE Nia Kurnianingsih; Juliyatin Putri Utami; Nurdiana Nurdiana; Diana Lyrawati
Indonesian Journal of Pharmacy Vol 27 No 3, 2016
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1107.374 KB) | DOI: 10.14499/indonesianjpharm27iss3pp128

Abstract

Excessive glutamate may give neurotoxic effects and contribute to Autism spectrum disorder(ASD). In this study, we investigated prolonged exposure effects of 10 µg/mL Monosodium Glutamate (MSG) on intracellular calcium level, bax, bcl-2, ratio of bax/bcl-2 genes expression, caspase-3, apoptosis of brain cells and stereotypic behavior of Zebrafish (Danio rerio) larvae at early developmental stages. Genes expression were determined by real time PCR, caspase-3 using ELISA, intracellular Ca2+ and apoptotic cells of brain using confocal microscopy, locomotor activity by using crossing lines assay whereas stereotypic behavior by circle swimming. The results indicated that MSG exposure increased brain bax and bcl-2; and caspase-3; intracellular Ca2+; and apoptosis; stereotypic behavior; and decreased locomotor activity. Termination of MSG treatments resulted in recovery of bax, bcl-2, caspase-3 basal levels and stereotypic behavior. In conclusion, MSG exposure at early embryonic stage increased brain cell damage and risk of behavior changes.
Catechins of GMB-4 Clone Inhibits Adipogenesis Through PPAR-γ and Adiponectin in Primary Culture of Visceral Preadipocyte of Rattus Norvegicus Wistar Aswaty Nur; Retty Ratnawati; Diana Lyrawati
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.
Co-Authors Alifia Putri Febriyanti Aswaty Nur Beta Herilla Sekti Debby Shintiya D Desie Suci Permata Sari Devi Anggraeni Pramulawati Djala, Fany Lairin Edi Widjajanto Elhemeidy, Rehab Mohammed Mustofa Ellen Natalia Faridha Khaira H Firman Mulyo Wicaksono Fitria Nurindro Rahmahani Habiba Aurora Herawati Sudoyo Herin Mawarti Herlina Y Handoko Hidayat Sujuti Indah Dina Maritha Juniartha, Putu Kenichi Yano Krisna Bayu Tambunan M Aris Widodo M. Rasjad Indra Mahfud, Rania Arif Martina Kurnia Rohmah Masruroh Rahayu, Masruroh Mohamad Rasjad Indra Mudjiwijono Handaru Muhammad Rasjad Indra Neshya Ruriana Putri, Neshya Ruriana Nia Kurnianingsih Noorma Lukitasari Novita Fahrianti Putri Nur Lailatul Fitria Nur Samsu Nurdiana Nurdiana Oktavia Rahayu Adianingsih Retty Ratnawati Retty Ratnawati Rudy Salam, Rudy Sangkot Marzuki Sarwono . Setyawati Soeharto, Setyawati Supranowo Supranowo Tatit Nurseta Tri Yudani Mardining Raras Utami, Juliyatin Putri Widjadjanto, Edi Yitania Sari