Homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells

AIM: To test whether homocysteine (Hcy) can influence the transcriptional profile, we hypothesized that Hcy can lead to the induction of proinflammatory molecules in the retinal cells of aging people. METHODS: An unbiased in vitro inflammatory pathway focused study was designed employing retinal pig...
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Gespeichert in:

Autor*in:	Mahavir Singh [verfasserIn] Suresh C Tyagi [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2017

Schlagwörter:	704 chemokine choroidal neovascularization cytokine gene expression hyperhomocysteinemia inflammation macular degeneration retinal remodeling

Übergeordnetes Werk:	In: International Journal of Ophthalmology - Press of International Journal of Ophthalmology (IJO PRESS), 2016, 10(2017), 5, Seite 696-704
Übergeordnetes Werk:	volume:10 ; year:2017 ; number:5 ; pages:696-704

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc Journal toc

DOI / URN:	10.18240/ijo.2017.05.06

Katalog-ID:	DOAJ044669127

Internformat


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520			\|a AIM: To test whether homocysteine (Hcy) can influence the transcriptional profile, we hypothesized that Hcy can lead to the induction of proinflammatory molecules in the retinal cells of aging people. METHODS: An unbiased in vitro inflammatory pathway focused study was designed employing retinal pigment epithelial (RPE) cell line, ARPE-19. Cells were cultured in the presence or absence of Hcy to capture target genes’ expression profile. Three different concentrations of Hcy were added in the culture medium of confluent monolayers. cRNAs were made from the isolated total RNAs and the labeled cRNA probes were hybridized to microarrays specific for human disease pathway inflammatory cytokines, chemokines and their receptor gene micro-array panels as per manufacture’s recommendations. Two Hcy up-regulated molecules: IL6 and CEBPB were further validated via Western blot analysis. Hcy’s effect on ARPE-19 cellular morphology and genomic DNA integrity were also evaluated. RESULTS: Gene microarray analyses of RPE cells in response to Hcy treatment revealed alterations in the expressions of several inflammatory gene transcripts such as CCL5, CEBPB, IL13RA2, IL15RA, IL6, IL8 and CXCL3 that were up-regulated. The transcripts for C3, CCL2, IL11RA and IL18 genes exhibited down-regulation. The IL6 and CEBPB expressions were subsequently validated at the protein levels. Treatment of the retinal cells with increasing Hcy concentration influenced their density in culture however their morphology and DNA integrity remained unaffected. CONCLUSION: These findings suggest that Hcy can potentially mediate the expression of chemokines, cytokines and interleukins receptors in the retinal cells without having any debilitating effects on their morphology and the genomic DNA integrity.
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author_variant	m s ms s c t sct
matchkey_str	article:22274898:2017----::ooytieeitsrncitoacagsfhifamtrptwyintrgnsnua
hierarchy_sort_str	2017
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allfields	10.18240/ijo.2017.05.06 doi (DE-627)DOAJ044669127 (DE-599)DOAJfdad84d56dbe4a4590bb8902a94aa204 DE-627 ger DE-627 rakwb eng RE1-994 Mahavir Singh verfasserin aut Homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier AIM: To test whether homocysteine (Hcy) can influence the transcriptional profile, we hypothesized that Hcy can lead to the induction of proinflammatory molecules in the retinal cells of aging people. METHODS: An unbiased in vitro inflammatory pathway focused study was designed employing retinal pigment epithelial (RPE) cell line, ARPE-19. Cells were cultured in the presence or absence of Hcy to capture target genes’ expression profile. Three different concentrations of Hcy were added in the culture medium of confluent monolayers. cRNAs were made from the isolated total RNAs and the labeled cRNA probes were hybridized to microarrays specific for human disease pathway inflammatory cytokines, chemokines and their receptor gene micro-array panels as per manufacture’s recommendations. Two Hcy up-regulated molecules: IL6 and CEBPB were further validated via Western blot analysis. Hcy’s effect on ARPE-19 cellular morphology and genomic DNA integrity were also evaluated. RESULTS: Gene microarray analyses of RPE cells in response to Hcy treatment revealed alterations in the expressions of several inflammatory gene transcripts such as CCL5, CEBPB, IL13RA2, IL15RA, IL6, IL8 and CXCL3 that were up-regulated. The transcripts for C3, CCL2, IL11RA and IL18 genes exhibited down-regulation. The IL6 and CEBPB expressions were subsequently validated at the protein levels. Treatment of the retinal cells with increasing Hcy concentration influenced their density in culture however their morphology and DNA integrity remained unaffected. CONCLUSION: These findings suggest that Hcy can potentially mediate the expression of chemokines, cytokines and interleukins receptors in the retinal cells without having any debilitating effects on their morphology and the genomic DNA integrity. 704 chemokine choroidal neovascularization cytokine gene expression hyperhomocysteinemia inflammation macular degeneration retinal remodeling Ophthalmology Suresh C Tyagi verfasserin aut In International Journal of Ophthalmology Press of International Journal of Ophthalmology (IJO PRESS), 2016 10(2017), 5, Seite 696-704 (DE-627)718635906 (DE-600)2663246-9 22274898 nnns volume:10 year:2017 number:5 pages:696-704 https://doi.org/10.18240/ijo.2017.05.06 kostenfrei https://doaj.org/article/fdad84d56dbe4a4590bb8902a94aa204 kostenfrei http://www.ijo.cn/en_publish/2017/5/20170506.pdf kostenfrei https://doaj.org/toc/2222-3959 Journal toc kostenfrei https://doaj.org/toc/2227-4898 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2817 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2017 5 696-704
spelling	10.18240/ijo.2017.05.06 doi (DE-627)DOAJ044669127 (DE-599)DOAJfdad84d56dbe4a4590bb8902a94aa204 DE-627 ger DE-627 rakwb eng RE1-994 Mahavir Singh verfasserin aut Homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier AIM: To test whether homocysteine (Hcy) can influence the transcriptional profile, we hypothesized that Hcy can lead to the induction of proinflammatory molecules in the retinal cells of aging people. METHODS: An unbiased in vitro inflammatory pathway focused study was designed employing retinal pigment epithelial (RPE) cell line, ARPE-19. Cells were cultured in the presence or absence of Hcy to capture target genes’ expression profile. Three different concentrations of Hcy were added in the culture medium of confluent monolayers. cRNAs were made from the isolated total RNAs and the labeled cRNA probes were hybridized to microarrays specific for human disease pathway inflammatory cytokines, chemokines and their receptor gene micro-array panels as per manufacture’s recommendations. Two Hcy up-regulated molecules: IL6 and CEBPB were further validated via Western blot analysis. Hcy’s effect on ARPE-19 cellular morphology and genomic DNA integrity were also evaluated. RESULTS: Gene microarray analyses of RPE cells in response to Hcy treatment revealed alterations in the expressions of several inflammatory gene transcripts such as CCL5, CEBPB, IL13RA2, IL15RA, IL6, IL8 and CXCL3 that were up-regulated. The transcripts for C3, CCL2, IL11RA and IL18 genes exhibited down-regulation. The IL6 and CEBPB expressions were subsequently validated at the protein levels. Treatment of the retinal cells with increasing Hcy concentration influenced their density in culture however their morphology and DNA integrity remained unaffected. CONCLUSION: These findings suggest that Hcy can potentially mediate the expression of chemokines, cytokines and interleukins receptors in the retinal cells without having any debilitating effects on their morphology and the genomic DNA integrity. 704 chemokine choroidal neovascularization cytokine gene expression hyperhomocysteinemia inflammation macular degeneration retinal remodeling Ophthalmology Suresh C Tyagi verfasserin aut In International Journal of Ophthalmology Press of International Journal of Ophthalmology (IJO PRESS), 2016 10(2017), 5, Seite 696-704 (DE-627)718635906 (DE-600)2663246-9 22274898 nnns volume:10 year:2017 number:5 pages:696-704 https://doi.org/10.18240/ijo.2017.05.06 kostenfrei https://doaj.org/article/fdad84d56dbe4a4590bb8902a94aa204 kostenfrei http://www.ijo.cn/en_publish/2017/5/20170506.pdf kostenfrei https://doaj.org/toc/2222-3959 Journal toc kostenfrei https://doaj.org/toc/2227-4898 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2817 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2017 5 696-704
allfields_unstemmed	10.18240/ijo.2017.05.06 doi (DE-627)DOAJ044669127 (DE-599)DOAJfdad84d56dbe4a4590bb8902a94aa204 DE-627 ger DE-627 rakwb eng RE1-994 Mahavir Singh verfasserin aut Homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier AIM: To test whether homocysteine (Hcy) can influence the transcriptional profile, we hypothesized that Hcy can lead to the induction of proinflammatory molecules in the retinal cells of aging people. METHODS: An unbiased in vitro inflammatory pathway focused study was designed employing retinal pigment epithelial (RPE) cell line, ARPE-19. Cells were cultured in the presence or absence of Hcy to capture target genes’ expression profile. Three different concentrations of Hcy were added in the culture medium of confluent monolayers. cRNAs were made from the isolated total RNAs and the labeled cRNA probes were hybridized to microarrays specific for human disease pathway inflammatory cytokines, chemokines and their receptor gene micro-array panels as per manufacture’s recommendations. Two Hcy up-regulated molecules: IL6 and CEBPB were further validated via Western blot analysis. Hcy’s effect on ARPE-19 cellular morphology and genomic DNA integrity were also evaluated. RESULTS: Gene microarray analyses of RPE cells in response to Hcy treatment revealed alterations in the expressions of several inflammatory gene transcripts such as CCL5, CEBPB, IL13RA2, IL15RA, IL6, IL8 and CXCL3 that were up-regulated. The transcripts for C3, CCL2, IL11RA and IL18 genes exhibited down-regulation. The IL6 and CEBPB expressions were subsequently validated at the protein levels. Treatment of the retinal cells with increasing Hcy concentration influenced their density in culture however their morphology and DNA integrity remained unaffected. CONCLUSION: These findings suggest that Hcy can potentially mediate the expression of chemokines, cytokines and interleukins receptors in the retinal cells without having any debilitating effects on their morphology and the genomic DNA integrity. 704 chemokine choroidal neovascularization cytokine gene expression hyperhomocysteinemia inflammation macular degeneration retinal remodeling Ophthalmology Suresh C Tyagi verfasserin aut In International Journal of Ophthalmology Press of International Journal of Ophthalmology (IJO PRESS), 2016 10(2017), 5, Seite 696-704 (DE-627)718635906 (DE-600)2663246-9 22274898 nnns volume:10 year:2017 number:5 pages:696-704 https://doi.org/10.18240/ijo.2017.05.06 kostenfrei https://doaj.org/article/fdad84d56dbe4a4590bb8902a94aa204 kostenfrei http://www.ijo.cn/en_publish/2017/5/20170506.pdf kostenfrei https://doaj.org/toc/2222-3959 Journal toc kostenfrei https://doaj.org/toc/2227-4898 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2817 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2017 5 696-704
allfieldsGer	10.18240/ijo.2017.05.06 doi (DE-627)DOAJ044669127 (DE-599)DOAJfdad84d56dbe4a4590bb8902a94aa204 DE-627 ger DE-627 rakwb eng RE1-994 Mahavir Singh verfasserin aut Homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier AIM: To test whether homocysteine (Hcy) can influence the transcriptional profile, we hypothesized that Hcy can lead to the induction of proinflammatory molecules in the retinal cells of aging people. METHODS: An unbiased in vitro inflammatory pathway focused study was designed employing retinal pigment epithelial (RPE) cell line, ARPE-19. Cells were cultured in the presence or absence of Hcy to capture target genes’ expression profile. Three different concentrations of Hcy were added in the culture medium of confluent monolayers. cRNAs were made from the isolated total RNAs and the labeled cRNA probes were hybridized to microarrays specific for human disease pathway inflammatory cytokines, chemokines and their receptor gene micro-array panels as per manufacture’s recommendations. Two Hcy up-regulated molecules: IL6 and CEBPB were further validated via Western blot analysis. Hcy’s effect on ARPE-19 cellular morphology and genomic DNA integrity were also evaluated. RESULTS: Gene microarray analyses of RPE cells in response to Hcy treatment revealed alterations in the expressions of several inflammatory gene transcripts such as CCL5, CEBPB, IL13RA2, IL15RA, IL6, IL8 and CXCL3 that were up-regulated. The transcripts for C3, CCL2, IL11RA and IL18 genes exhibited down-regulation. The IL6 and CEBPB expressions were subsequently validated at the protein levels. Treatment of the retinal cells with increasing Hcy concentration influenced their density in culture however their morphology and DNA integrity remained unaffected. CONCLUSION: These findings suggest that Hcy can potentially mediate the expression of chemokines, cytokines and interleukins receptors in the retinal cells without having any debilitating effects on their morphology and the genomic DNA integrity. 704 chemokine choroidal neovascularization cytokine gene expression hyperhomocysteinemia inflammation macular degeneration retinal remodeling Ophthalmology Suresh C Tyagi verfasserin aut In International Journal of Ophthalmology Press of International Journal of Ophthalmology (IJO PRESS), 2016 10(2017), 5, Seite 696-704 (DE-627)718635906 (DE-600)2663246-9 22274898 nnns volume:10 year:2017 number:5 pages:696-704 https://doi.org/10.18240/ijo.2017.05.06 kostenfrei https://doaj.org/article/fdad84d56dbe4a4590bb8902a94aa204 kostenfrei http://www.ijo.cn/en_publish/2017/5/20170506.pdf kostenfrei https://doaj.org/toc/2222-3959 Journal toc kostenfrei https://doaj.org/toc/2227-4898 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2817 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2017 5 696-704
allfieldsSound	10.18240/ijo.2017.05.06 doi (DE-627)DOAJ044669127 (DE-599)DOAJfdad84d56dbe4a4590bb8902a94aa204 DE-627 ger DE-627 rakwb eng RE1-994 Mahavir Singh verfasserin aut Homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier AIM: To test whether homocysteine (Hcy) can influence the transcriptional profile, we hypothesized that Hcy can lead to the induction of proinflammatory molecules in the retinal cells of aging people. METHODS: An unbiased in vitro inflammatory pathway focused study was designed employing retinal pigment epithelial (RPE) cell line, ARPE-19. Cells were cultured in the presence or absence of Hcy to capture target genes’ expression profile. Three different concentrations of Hcy were added in the culture medium of confluent monolayers. cRNAs were made from the isolated total RNAs and the labeled cRNA probes were hybridized to microarrays specific for human disease pathway inflammatory cytokines, chemokines and their receptor gene micro-array panels as per manufacture’s recommendations. Two Hcy up-regulated molecules: IL6 and CEBPB were further validated via Western blot analysis. Hcy’s effect on ARPE-19 cellular morphology and genomic DNA integrity were also evaluated. RESULTS: Gene microarray analyses of RPE cells in response to Hcy treatment revealed alterations in the expressions of several inflammatory gene transcripts such as CCL5, CEBPB, IL13RA2, IL15RA, IL6, IL8 and CXCL3 that were up-regulated. The transcripts for C3, CCL2, IL11RA and IL18 genes exhibited down-regulation. The IL6 and CEBPB expressions were subsequently validated at the protein levels. Treatment of the retinal cells with increasing Hcy concentration influenced their density in culture however their morphology and DNA integrity remained unaffected. CONCLUSION: These findings suggest that Hcy can potentially mediate the expression of chemokines, cytokines and interleukins receptors in the retinal cells without having any debilitating effects on their morphology and the genomic DNA integrity. 704 chemokine choroidal neovascularization cytokine gene expression hyperhomocysteinemia inflammation macular degeneration retinal remodeling Ophthalmology Suresh C Tyagi verfasserin aut In International Journal of Ophthalmology Press of International Journal of Ophthalmology (IJO PRESS), 2016 10(2017), 5, Seite 696-704 (DE-627)718635906 (DE-600)2663246-9 22274898 nnns volume:10 year:2017 number:5 pages:696-704 https://doi.org/10.18240/ijo.2017.05.06 kostenfrei https://doaj.org/article/fdad84d56dbe4a4590bb8902a94aa204 kostenfrei http://www.ijo.cn/en_publish/2017/5/20170506.pdf kostenfrei https://doaj.org/toc/2222-3959 Journal toc kostenfrei https://doaj.org/toc/2227-4898 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2817 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2017 5 696-704
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source	In International Journal of Ophthalmology 10(2017), 5, Seite 696-704 volume:10 year:2017 number:5 pages:696-704
sourceStr	In International Journal of Ophthalmology 10(2017), 5, Seite 696-704 volume:10 year:2017 number:5 pages:696-704
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topic_facet	704 chemokine choroidal neovascularization cytokine gene expression hyperhomocysteinemia inflammation macular degeneration retinal remodeling Ophthalmology
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callnumber-first	R - Medicine
author	Mahavir Singh
spellingShingle	Mahavir Singh misc RE1-994 misc 704 misc chemokine misc choroidal neovascularization misc cytokine misc gene expression misc hyperhomocysteinemia misc inflammation misc macular degeneration misc retinal remodeling misc Ophthalmology Homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells
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topic_title	RE1-994 Homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells 704 chemokine choroidal neovascularization cytokine gene expression hyperhomocysteinemia inflammation macular degeneration retinal remodeling
topic	misc RE1-994 misc 704 misc chemokine misc choroidal neovascularization misc cytokine misc gene expression misc hyperhomocysteinemia misc inflammation misc macular degeneration misc retinal remodeling misc Ophthalmology
topic_unstemmed	misc RE1-994 misc 704 misc chemokine misc choroidal neovascularization misc cytokine misc gene expression misc hyperhomocysteinemia misc inflammation misc macular degeneration misc retinal remodeling misc Ophthalmology
topic_browse	misc RE1-994 misc 704 misc chemokine misc choroidal neovascularization misc cytokine misc gene expression misc hyperhomocysteinemia misc inflammation misc macular degeneration misc retinal remodeling misc Ophthalmology
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title	Homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells
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title_full	Homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells
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title_sort	homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells
callnumber	RE1-994
title_auth	Homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells
abstract	AIM: To test whether homocysteine (Hcy) can influence the transcriptional profile, we hypothesized that Hcy can lead to the induction of proinflammatory molecules in the retinal cells of aging people. METHODS: An unbiased in vitro inflammatory pathway focused study was designed employing retinal pigment epithelial (RPE) cell line, ARPE-19. Cells were cultured in the presence or absence of Hcy to capture target genes’ expression profile. Three different concentrations of Hcy were added in the culture medium of confluent monolayers. cRNAs were made from the isolated total RNAs and the labeled cRNA probes were hybridized to microarrays specific for human disease pathway inflammatory cytokines, chemokines and their receptor gene micro-array panels as per manufacture’s recommendations. Two Hcy up-regulated molecules: IL6 and CEBPB were further validated via Western blot analysis. Hcy’s effect on ARPE-19 cellular morphology and genomic DNA integrity were also evaluated. RESULTS: Gene microarray analyses of RPE cells in response to Hcy treatment revealed alterations in the expressions of several inflammatory gene transcripts such as CCL5, CEBPB, IL13RA2, IL15RA, IL6, IL8 and CXCL3 that were up-regulated. The transcripts for C3, CCL2, IL11RA and IL18 genes exhibited down-regulation. The IL6 and CEBPB expressions were subsequently validated at the protein levels. Treatment of the retinal cells with increasing Hcy concentration influenced their density in culture however their morphology and DNA integrity remained unaffected. CONCLUSION: These findings suggest that Hcy can potentially mediate the expression of chemokines, cytokines and interleukins receptors in the retinal cells without having any debilitating effects on their morphology and the genomic DNA integrity.
abstractGer	AIM: To test whether homocysteine (Hcy) can influence the transcriptional profile, we hypothesized that Hcy can lead to the induction of proinflammatory molecules in the retinal cells of aging people. METHODS: An unbiased in vitro inflammatory pathway focused study was designed employing retinal pigment epithelial (RPE) cell line, ARPE-19. Cells were cultured in the presence or absence of Hcy to capture target genes’ expression profile. Three different concentrations of Hcy were added in the culture medium of confluent monolayers. cRNAs were made from the isolated total RNAs and the labeled cRNA probes were hybridized to microarrays specific for human disease pathway inflammatory cytokines, chemokines and their receptor gene micro-array panels as per manufacture’s recommendations. Two Hcy up-regulated molecules: IL6 and CEBPB were further validated via Western blot analysis. Hcy’s effect on ARPE-19 cellular morphology and genomic DNA integrity were also evaluated. RESULTS: Gene microarray analyses of RPE cells in response to Hcy treatment revealed alterations in the expressions of several inflammatory gene transcripts such as CCL5, CEBPB, IL13RA2, IL15RA, IL6, IL8 and CXCL3 that were up-regulated. The transcripts for C3, CCL2, IL11RA and IL18 genes exhibited down-regulation. The IL6 and CEBPB expressions were subsequently validated at the protein levels. Treatment of the retinal cells with increasing Hcy concentration influenced their density in culture however their morphology and DNA integrity remained unaffected. CONCLUSION: These findings suggest that Hcy can potentially mediate the expression of chemokines, cytokines and interleukins receptors in the retinal cells without having any debilitating effects on their morphology and the genomic DNA integrity.
abstract_unstemmed	AIM: To test whether homocysteine (Hcy) can influence the transcriptional profile, we hypothesized that Hcy can lead to the induction of proinflammatory molecules in the retinal cells of aging people. METHODS: An unbiased in vitro inflammatory pathway focused study was designed employing retinal pigment epithelial (RPE) cell line, ARPE-19. Cells were cultured in the presence or absence of Hcy to capture target genes’ expression profile. Three different concentrations of Hcy were added in the culture medium of confluent monolayers. cRNAs were made from the isolated total RNAs and the labeled cRNA probes were hybridized to microarrays specific for human disease pathway inflammatory cytokines, chemokines and their receptor gene micro-array panels as per manufacture’s recommendations. Two Hcy up-regulated molecules: IL6 and CEBPB were further validated via Western blot analysis. Hcy’s effect on ARPE-19 cellular morphology and genomic DNA integrity were also evaluated. RESULTS: Gene microarray analyses of RPE cells in response to Hcy treatment revealed alterations in the expressions of several inflammatory gene transcripts such as CCL5, CEBPB, IL13RA2, IL15RA, IL6, IL8 and CXCL3 that were up-regulated. The transcripts for C3, CCL2, IL11RA and IL18 genes exhibited down-regulation. The IL6 and CEBPB expressions were subsequently validated at the protein levels. Treatment of the retinal cells with increasing Hcy concentration influenced their density in culture however their morphology and DNA integrity remained unaffected. CONCLUSION: These findings suggest that Hcy can potentially mediate the expression of chemokines, cytokines and interleukins receptors in the retinal cells without having any debilitating effects on their morphology and the genomic DNA integrity.
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title_short	Homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells
url	https://doi.org/10.18240/ijo.2017.05.06 https://doaj.org/article/fdad84d56dbe4a4590bb8902a94aa204 http://www.ijo.cn/en_publish/2017/5/20170506.pdf https://doaj.org/toc/2222-3959 https://doaj.org/toc/2227-4898
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up_date	2024-07-04T00:01:23.808Z
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Homocysteine mediates transcriptional changes of the inflammatory pathway signature genes in human retinal pigment epithelial cells

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