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Decreasing a-synuclein aggregation by methanolic extract of Centella asiatica in zebrafish Parkinson's model

Asian Pacific Journal of Tropical Biomedicine 2015年第11期5卷 948-954页

作者： Husnul Khotimah Mulyohadi Ali Sutiman Bambang Sumitro mochamad aris widodo Medical Faculty Brawijaya University Laboratory of Pharmacology Medical FacultyBrawijaya University Department of Biology Faculty of ScienceBrawijaya University

Objective: To observe the effects of Centella asiatica(C. asiatica) methanolic extract on a-synuclein aggregation and its expression in rotenone-exposed zebra fish.Methods: Zebra fish(Danio rerio) were exposed to 5 m g/L rotenone for 28 days and coincubated with 2.5, 5.0 and 10.0 m g/mL of C. asiatica methanolic extract. The medium was changed every 48 h for maintain the concentration of rotenone and extract. After 28 days zebra fish were sacrificed on the ice block and protein was isolated from zebra fish brain for ELISA of dopamine and Western blotting of a-synuclein. Immunohistochemistry was conducted to observe the a-synuclein expressions from histopathological preparation of zebra fish brain. The head were soaked in 10% formaline for less than 24 h and embedded onto paraffin block, then sliced for immunohistochemistry using anti a-synuclein antibody. We also measured zebra fish motility for 5 min in each week.Results: C. asiatica has important bioactive compounds such as asiaticoside that has antiin flammatory and antioxidant properties. It may inhibit cascade reaction due to oxidative stress induced by rotenone. Decreasing reactive oxygen species proposed probability of radical attack to a-synuclein protein that caused aggregation and increase of its expression.The motility of zebra fish was also maintained in C. asiatica groups due to the increasing dopamine level in rotenone-induced zebra fish. High level of reactive oxygen species inactivated enzyme for dopamine synthesis such as tyrosine hydroxylase, and oxidized dopamine itself. Oxidized dopamine increased a-synuclein aggregation. Thus, the dopamine level decreased in rotenone-induced zebra fish, but C. asiatica increased dopamine level.Conclusions: C. asiatica has a potential to be developed as an anti-Parkinson's disease treatment due to its capability for minimized the sign of Parkinson's such as a-synuclein aggregation and expression, increasing motility and dopamine as well.

关键词： a-Synuclein Centella asiatica Rotenone Parkinson’s

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Mechanism of retinal pericyte migration through Angiopoietin/Tie-2 signaling pathway on diabetic rats

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International Journal of Ophthalmology(English edition) 2018年第3期11卷 375-381页

作者： Nadia Artha Dewi Aulanni'am Aulanni 'am Hidayat Sujuti Muhammad aris widodo Ojoko Wahono Soeatmadji Department of Ophthalmology Vitreoretinal Subdivision Faculty of Medicine Brawijaya University Malang 65111 Indonesia Department of Biochemistry Faculty of Sciences Brawijaya University Malang 65111 Indonesia Department of Biochemistry Faculty of Medicine Brawijaya University Malang 65111 Indonesia Department of Pharmacology Faculty of Medicine Brawijaya University Malang 65111 Indonesia Department of Endocrinology Faculty of Medicine Brawijaya University Malang 65111 Indonesia

AIM： To investigate the mechanism of pericyte migration through Angiopoietin-2 （Ang-2）/Tie-2 signaling pathway. METHODS： We divided the rats into 5 groups. Each diabetic rat model groups injected with Tie-2 inhibitor, ERK1/2 inhibitor, Akt/PKB inhibitor, and DMSO intravitreal. Retinal digest preparation was done to examine the retinal vasculature including pericyte： endothelial ratio, and morphology of pericyte migration. Tie-2, ERKI/2 and Akt/PKB phosporylation were analyzed by confocal laser scanning microscopy. RESULTS： There was a correlation between pericyte migration with increasing Ang-2 （P〈0.05）. Pericyte number reduced by 40% （1：2.4） after 5wk diabetes on diabetic rats. The pericyte： endothelial ratio on group with Tie-2 inhibitor were 1：1.8. The same result shows on group with Akt/PKB inhibition. ERK1/2 inhibitor group shows the best results of pericyte： endothelial ratio （1：1,7）, Inhibition on Tie-2 receptor decreased the phosphorylation activity of Tie-2, ERK1/2 and Akt/PKB pathway. ERK1/2 inhibition also decreasing the phosphorylation of Tie-2 and Akt/PKB. But on Akt/PKB inhibition, the phosphorylation of Tie-2 and ERK1/2 were relative the same. CONCLUSION： Ang-2 has a role for pericyte migration on diabetic rats through Tie-2 receptor, ERKII2 and Akt/PKB pathways. ERK1/2 is a dominant pathway based on the ability to supress another pathway activity and decreasing pericyte migration on diabetic rats.

关键词： pericyte Angiopoietin/Tie-2 diabetes cellmigration rat

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Neuropathy optic glaucomatosa induced by systemic hypertension through activation endothelin-1 signaling pathway in central retinal artery in rats

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International Journal of Ophthalmology(English edition) 2016年第11期9卷 1568-1577页

作者： Seskoati Prayitnaningsih Hidayat Sujuti Maksum Effendi Aulia Abdullah Nanda Wahyu Anandita Febriani Yohana Nur Permatasari Mohamad aris widodo Department of Ophthalmology Faculty of MedicineBrawijaya University Pharmacology Laboratory Faculty of MedicineBrawijaya University

AIM： To evaluate effect of hypertension on retinal ganglion cell（RGC） apoptosis, intraocular pressure（IOP）,and the activation of endothelin-1（ET-1） signaling pathway in central retinal artery（CRA） in rats.METHODS： The experimental study was performed on20 male Sprague Dawley rats that were divided into control group, and hypertension groups. The hypertension was induced by subcutaneous deoxycorticoacetate（DOCA）10 mg/kg twice a week and administered 0.9% Na Cl solution daily for 2, 6, and 10 wk. Blood pressure（BP） was measured using animal BP analyzer. IOP was measured by handheld tonometry. Retinal tissue preparations by paraffin blocks were made after enucleation. The expression of ET-1, e NOS, ET-1 receptor A（ETRA）, ET-1receptor B（ETRB）, and phosphorylated myosin light chain kinase（MLCK）, and caldesmon（Ca D） in CRA and RGC apoptosis were evaluated through immunofluorescent staining method then observed using laser scanning confocal microscopy. RESULTS： BP significantly increased in all of the hypertension groups compared to control（P =0.001）.Peak IOP elevation（7.78±4.14 mm Hg） and RGC apoptosis（576.15±33.28 Au） occurred on 2wk of hypertension. ET-1expression（1238.6±55.1 Au） and e NOS expression（2814.2±70.7 Au） were found highest in 2wk of hypertension,although the ratio of ET-1/e NOS decreased since 2wk.ETRAreached peak expression in 10 wk of hypertension（1219.4 ±6.3 Au）, while ETRBsignificantly increased only in 2 weeks group（1069.2 ±9.6 Au）. The highest MLCK expression（1190.09±58.32 Au）, Ca D（1670.28±18.36 Au）were also found in 2wk of hypertension.CONCLUSION： Hypertension effects to activation of ET-1 signaling pathway significantly in CRA, elevation of IOP, and RGC apoptosis. The highest value was achieved at 2wk, which is the development phase of hypertension.

关键词： hypertension central retinal artery endothelin- 1 signaling pathway rats

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Anti-diabetic potential of Urena lobata leaf extract through inhibition of dipeptidyl peptidase IV activity

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Asian Pacific Journal of Tropical Biomedicine 2015年第8期5卷 630-634页

作者： Yudi Purnomo Djoko Wahono Soeatmadji Sutiman Bambang Sumitro Mochammad aris widodo School of Medicine Brawijaya University Pharmacology Department School of Medicine Malang Islamic University Internal Department School of Medicine Brawijaya University Biology Department Faculty of Science Brawijaya University Pharmacology Department School of Medicine Brawijaya University

Objective: To evaluate the anti-diabetic potential of leaf extract from Urena lobata(U. lobata) through dipeptidyl peptidase IV(DPP-IV) inhibitory activity.Methods: U. lobata leaf was extracted in hot water and ethanol. The activity of DPPIV inhibitor was tested by in vitro study using gly-pro-p-nitroanilide as substrat of DPPIV and vildagliptin, as standard reference. A product of the reactions between gly-pro-pnitroanilide and DPP-IV, was observed by microplate readers with λ = 405 nm. All data were expressed as mean ± SD and the IC50 value was determined by non linear regression curve fit. Active substances in leaf extract of U. lobata was analyzed by liquid chromatography-mass spectrometry. DPP-IV inhibitory activity of active compounds was evaluated in silico using docking server. Results: The ethanolic extract of U. lobata showed stronger DPP-IV inhibitor activity than water extract with the IC50 values of 1 654.64 and 6 489.88 μg/mL, respectively. Vildagliptin, based on standard reference for DPP-IV inhibitor activity, has IC50 value of 57.44 μg/mL. Based on in silico analysis, mangiferin, stigmasterol and β-sitosterol in U. lobata extract have a strong inhibitory activity on DPP-IV. Conclusions: The results showed that DPP-IV inhibitory activity of U. lobata is related to its active compounds such as mangiferin, stigmasterol and β-sitosterol.

关键词： Anti-diabetic Dipeptidyl peptidase IV In silico In vitro Urena lobata

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