Adipose Tissue‐Derived Stem Cells Secrete CXCL5 Cytokine with Neurotrophic Effects on Cavernous Nerve Regeneration

Previously we reported that paracrine actions likely mediated the therapeutic effects of adipose tissue‐derived stem cells (ADSCs) on a rat model of cavernous nerve (CN) injury. To identify potential neurotrophic factors in ADSC's secretion, test the most promising one, and identify the molecul...

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Veröffentlicht in:Journal of sexual medicine 2011-02, Vol.8 (2), p.437-446
Hauptverfasser: Zhang, Haiyang, Yang, Rong, Wang, Zhong, Lin, Guiting, Lue, Tom F., Lin, Ching‐Shwun
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container_issue 2
container_start_page 437
container_title Journal of sexual medicine
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creator Zhang, Haiyang
Yang, Rong
Wang, Zhong
Lin, Guiting
Lue, Tom F.
Lin, Ching‐Shwun
description Previously we reported that paracrine actions likely mediated the therapeutic effects of adipose tissue‐derived stem cells (ADSCs) on a rat model of cavernous nerve (CN) injury. To identify potential neurotrophic factors in ADSC's secretion, test the most promising one, and identify the molecular mechanism of its neurotrophic action. Rat major pelvic ganglia (MPG) were cultured in conditioned media of ADSC and penile smooth muscle cells (PSMCs). Cytokine expression in these two media was probed with a cytokine antibody array. CXCL5 cytokine was quantified in these two media by enzyme‐linked immunosorbent assay (ELISA). Activation of Janus Kinase/Signal Transducer and Activator of Transcription (JAK/STAT) by CXCL5 was tested in neuroblastoma cell lines BE(2)C and SH‐SY5Y as well as in Schwann cell line RT4‐D6P2T by Western blot. Involvement of CXCL5 and JAK/STAT in ADSC‐conditioned medium's neurotrophic effects was confirmed with anti‐CXCL5 antibody and JAK inhibitor AG490, respectively. Neurotrophic effects of ADSC and PSMC‐conditioned media were quantified by measuring neurite length in MPG cultures. Secretion of CXCL5 in these two media was quantified by ELISA. Activation of JAK/STAT by CXCL5 was quantified by densitometry on Western blots for STAT1 and STAT3 phosphorylation. MPG neurite length was significantly longer in ADSC than in PSMC‐conditioned medium. CXCL5 was secreted eight times higher in ADSC than in PSMC‐conditioned medium. Anti‐CXCL5 antibody blocked the neurotrophic effects of ADSC‐conditioned medium. CXCL5 activated JAK/STAT concentration‐dependently from 0 to 50 ng/mL in RT4‐D6P2T Schwann cells. At 50 ng/mL, CXCL5 activated JAK/STAT time‐dependently, peaking at 45 minutes. AG490 blocked these activities as well as the neurotrophic effects of ADSC‐conditioned medium. CXCL5 was secreted by ADSC at a high level, promoted MPG neurite growth, and activated JAK/STAT in Schwann cells. CXCL5 may contribute to ADSC's therapeutic efficacy on CN injury‐induced ED. Zhang H, Yang R, Wang Z, Lin G, Lue TF, and Lin C‐S. Adipose tissue‐derived stem cells secrete CXCL5 cytokine with neurotrophic effects on cavernous nerve regeneration. J Sex Med 2011;8:437–446.
doi_str_mv 10.1111/j.1743-6109.2010.02128.x
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To identify potential neurotrophic factors in ADSC's secretion, test the most promising one, and identify the molecular mechanism of its neurotrophic action. Rat major pelvic ganglia (MPG) were cultured in conditioned media of ADSC and penile smooth muscle cells (PSMCs). Cytokine expression in these two media was probed with a cytokine antibody array. CXCL5 cytokine was quantified in these two media by enzyme‐linked immunosorbent assay (ELISA). Activation of Janus Kinase/Signal Transducer and Activator of Transcription (JAK/STAT) by CXCL5 was tested in neuroblastoma cell lines BE(2)C and SH‐SY5Y as well as in Schwann cell line RT4‐D6P2T by Western blot. Involvement of CXCL5 and JAK/STAT in ADSC‐conditioned medium's neurotrophic effects was confirmed with anti‐CXCL5 antibody and JAK inhibitor AG490, respectively. Neurotrophic effects of ADSC and PSMC‐conditioned media were quantified by measuring neurite length in MPG cultures. Secretion of CXCL5 in these two media was quantified by ELISA. Activation of JAK/STAT by CXCL5 was quantified by densitometry on Western blots for STAT1 and STAT3 phosphorylation. MPG neurite length was significantly longer in ADSC than in PSMC‐conditioned medium. CXCL5 was secreted eight times higher in ADSC than in PSMC‐conditioned medium. Anti‐CXCL5 antibody blocked the neurotrophic effects of ADSC‐conditioned medium. CXCL5 activated JAK/STAT concentration‐dependently from 0 to 50 ng/mL in RT4‐D6P2T Schwann cells. At 50 ng/mL, CXCL5 activated JAK/STAT time‐dependently, peaking at 45 minutes. AG490 blocked these activities as well as the neurotrophic effects of ADSC‐conditioned medium. CXCL5 was secreted by ADSC at a high level, promoted MPG neurite growth, and activated JAK/STAT in Schwann cells. CXCL5 may contribute to ADSC's therapeutic efficacy on CN injury‐induced ED. Zhang H, Yang R, Wang Z, Lin G, Lue TF, and Lin C‐S. Adipose tissue‐derived stem cells secrete CXCL5 cytokine with neurotrophic effects on cavernous nerve regeneration. J Sex Med 2011;8:437–446.</description><identifier>ISSN: 1743-6095</identifier><identifier>EISSN: 1743-6109</identifier><identifier>DOI: 10.1111/j.1743-6109.2010.02128.x</identifier><identifier>PMID: 21114767</identifier><language>eng</language><publisher>Malden, USA: Elsevier Inc</publisher><subject>Adipose Tissue - cytology ; Adipose Tissue-Derived Stem Cells ; Animals ; Antibodies ; Blotting, Western ; Cavernous Nerve Regeneration ; Cell Line, Tumor ; Cells, Cultured ; Chemokine CXCL5 - pharmacology ; Chemokine CXCL5 - physiology ; CXCL5 Cytokine ; Enzyme Activation - drug effects ; Enzyme-Linked Immunosorbent Assay ; Erectile Dysfunction ; JAK/STAT ; Janus Kinases - metabolism ; Male ; Muscle, Smooth, Vascular - innervation ; Muscle, Smooth, Vascular - physiology ; Nerve Regeneration - drug effects ; Nerve Regeneration - physiology ; Neurites - drug effects ; Neurites - physiology ; Penis - innervation ; Penis - physiology ; Rats ; Rats, Sprague-Dawley ; Schwann Cells - drug effects ; Schwann Cells - physiology ; Signal Transduction - drug effects ; Signal Transduction - physiology ; STAT Transcription Factors - metabolism ; Stem Cells - physiology ; Stem Cells - secretion</subject><ispartof>Journal of sexual medicine, 2011-02, Vol.8 (2), p.437-446</ispartof><rights>2011 International Society for Sexual Medicine</rights><rights>2010 International Society for Sexual Medicine</rights><rights>2010 International Society for Sexual Medicine.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c6028-1444cbe0ab6430f1e1a05ef99ce842ef8d7ade06a296585edf6225656f7d875c3</citedby><cites>FETCH-LOGICAL-c6028-1444cbe0ab6430f1e1a05ef99ce842ef8d7ade06a296585edf6225656f7d875c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fj.1743-6109.2010.02128.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1743-6109.2010.02128.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>230,314,780,784,885,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21114767$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhang, Haiyang</creatorcontrib><creatorcontrib>Yang, Rong</creatorcontrib><creatorcontrib>Wang, Zhong</creatorcontrib><creatorcontrib>Lin, Guiting</creatorcontrib><creatorcontrib>Lue, Tom F.</creatorcontrib><creatorcontrib>Lin, Ching‐Shwun</creatorcontrib><title>Adipose Tissue‐Derived Stem Cells Secrete CXCL5 Cytokine with Neurotrophic Effects on Cavernous Nerve Regeneration</title><title>Journal of sexual medicine</title><addtitle>J Sex Med</addtitle><description>Previously we reported that paracrine actions likely mediated the therapeutic effects of adipose tissue‐derived stem cells (ADSCs) on a rat model of cavernous nerve (CN) injury. To identify potential neurotrophic factors in ADSC's secretion, test the most promising one, and identify the molecular mechanism of its neurotrophic action. Rat major pelvic ganglia (MPG) were cultured in conditioned media of ADSC and penile smooth muscle cells (PSMCs). Cytokine expression in these two media was probed with a cytokine antibody array. CXCL5 cytokine was quantified in these two media by enzyme‐linked immunosorbent assay (ELISA). Activation of Janus Kinase/Signal Transducer and Activator of Transcription (JAK/STAT) by CXCL5 was tested in neuroblastoma cell lines BE(2)C and SH‐SY5Y as well as in Schwann cell line RT4‐D6P2T by Western blot. Involvement of CXCL5 and JAK/STAT in ADSC‐conditioned medium's neurotrophic effects was confirmed with anti‐CXCL5 antibody and JAK inhibitor AG490, respectively. Neurotrophic effects of ADSC and PSMC‐conditioned media were quantified by measuring neurite length in MPG cultures. Secretion of CXCL5 in these two media was quantified by ELISA. Activation of JAK/STAT by CXCL5 was quantified by densitometry on Western blots for STAT1 and STAT3 phosphorylation. MPG neurite length was significantly longer in ADSC than in PSMC‐conditioned medium. CXCL5 was secreted eight times higher in ADSC than in PSMC‐conditioned medium. Anti‐CXCL5 antibody blocked the neurotrophic effects of ADSC‐conditioned medium. CXCL5 activated JAK/STAT concentration‐dependently from 0 to 50 ng/mL in RT4‐D6P2T Schwann cells. At 50 ng/mL, CXCL5 activated JAK/STAT time‐dependently, peaking at 45 minutes. AG490 blocked these activities as well as the neurotrophic effects of ADSC‐conditioned medium. CXCL5 was secreted by ADSC at a high level, promoted MPG neurite growth, and activated JAK/STAT in Schwann cells. CXCL5 may contribute to ADSC's therapeutic efficacy on CN injury‐induced ED. Zhang H, Yang R, Wang Z, Lin G, Lue TF, and Lin C‐S. Adipose tissue‐derived stem cells secrete CXCL5 cytokine with neurotrophic effects on cavernous nerve regeneration. 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To identify potential neurotrophic factors in ADSC's secretion, test the most promising one, and identify the molecular mechanism of its neurotrophic action. Rat major pelvic ganglia (MPG) were cultured in conditioned media of ADSC and penile smooth muscle cells (PSMCs). Cytokine expression in these two media was probed with a cytokine antibody array. CXCL5 cytokine was quantified in these two media by enzyme‐linked immunosorbent assay (ELISA). Activation of Janus Kinase/Signal Transducer and Activator of Transcription (JAK/STAT) by CXCL5 was tested in neuroblastoma cell lines BE(2)C and SH‐SY5Y as well as in Schwann cell line RT4‐D6P2T by Western blot. Involvement of CXCL5 and JAK/STAT in ADSC‐conditioned medium's neurotrophic effects was confirmed with anti‐CXCL5 antibody and JAK inhibitor AG490, respectively. Neurotrophic effects of ADSC and PSMC‐conditioned media were quantified by measuring neurite length in MPG cultures. Secretion of CXCL5 in these two media was quantified by ELISA. Activation of JAK/STAT by CXCL5 was quantified by densitometry on Western blots for STAT1 and STAT3 phosphorylation. MPG neurite length was significantly longer in ADSC than in PSMC‐conditioned medium. CXCL5 was secreted eight times higher in ADSC than in PSMC‐conditioned medium. Anti‐CXCL5 antibody blocked the neurotrophic effects of ADSC‐conditioned medium. CXCL5 activated JAK/STAT concentration‐dependently from 0 to 50 ng/mL in RT4‐D6P2T Schwann cells. At 50 ng/mL, CXCL5 activated JAK/STAT time‐dependently, peaking at 45 minutes. AG490 blocked these activities as well as the neurotrophic effects of ADSC‐conditioned medium. CXCL5 was secreted by ADSC at a high level, promoted MPG neurite growth, and activated JAK/STAT in Schwann cells. CXCL5 may contribute to ADSC's therapeutic efficacy on CN injury‐induced ED. Zhang H, Yang R, Wang Z, Lin G, Lue TF, and Lin C‐S. Adipose tissue‐derived stem cells secrete CXCL5 cytokine with neurotrophic effects on cavernous nerve regeneration. J Sex Med 2011;8:437–446.</abstract><cop>Malden, USA</cop><pub>Elsevier Inc</pub><pmid>21114767</pmid><doi>10.1111/j.1743-6109.2010.02128.x</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record>
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subjects Adipose Tissue - cytology
Adipose Tissue-Derived Stem Cells
Animals
Antibodies
Blotting, Western
Cavernous Nerve Regeneration
Cell Line, Tumor
Cells, Cultured
Chemokine CXCL5 - pharmacology
Chemokine CXCL5 - physiology
CXCL5 Cytokine
Enzyme Activation - drug effects
Enzyme-Linked Immunosorbent Assay
Erectile Dysfunction
JAK/STAT
Janus Kinases - metabolism
Male
Muscle, Smooth, Vascular - innervation
Muscle, Smooth, Vascular - physiology
Nerve Regeneration - drug effects
Nerve Regeneration - physiology
Neurites - drug effects
Neurites - physiology
Penis - innervation
Penis - physiology
Rats
Rats, Sprague-Dawley
Schwann Cells - drug effects
Schwann Cells - physiology
Signal Transduction - drug effects
Signal Transduction - physiology
STAT Transcription Factors - metabolism
Stem Cells - physiology
Stem Cells - secretion
title Adipose Tissue‐Derived Stem Cells Secrete CXCL5 Cytokine with Neurotrophic Effects on Cavernous Nerve Regeneration
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