Expression and regulation of neurotrophic and angiogenic factors during human intervertebral disc degeneration

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1、LABinchetal.ArthritisResearch&Therapy2014,16:416http://arthritis-research.com/content/16/4/416RESEARCHARTICLEOpenAccessExpressionandregulationofneurotrophicandangiogenicfactorsduringhumanintervertebraldiscdegeneration122221AbbieLABinch,AshleyACole,LeeMBreakwell,AnthonyLRMichael,NeilC

2、hiverton,AlisonKCross1*andChristineLLeMaitreAbstractIntroduction:Thedegenerateintervertebraldisc(IVD)becomesinnervatedbysensorynervefibres,andvascularisedbybloodvessels.Thisstudyaimedtoidentifyneurotrophins,neuropeptidesandangiogenicfactorswithinnativeIVDtissueandtofurtherinvestigate

3、whetherpro-inflammatorycytokinesareinvolvedintheregulationofexpressionlevelswithinnucleuspulposus(NP)cells,nerveandendothelialcells.Methods:Quantitativereal-timePCR(qRT-PCR)wasperformedon53humanIVDsfrom52individualstoinvestigatenativegeneexpressionofneurotrophicfactorsandtheirrecepto

4、rs,neuropeptidesandangiogenicfactors.TheregulationofthesefactorsbycytokineswasinvestigatedinNPcellsinalginateculture,andnerveandendothelialcellsinmonolayerusingRT-PCRandsubstanceP(SP)proteinexpressionininterleukin-1(IL-1β)stimulatedNPcells.Results:InitialinvestigationonunculturedNPce

5、llsidentifiedexpressionofallneurotrophinsbynativeNPcells,whilstthenervegrowthfactor(NGF)receptorwasonlyidentifiedinseverelydegenerateandinfiltrateddiscs,andbrainderivedneurotrophicfactor(BDNF)receptorexpressedbymoredegeneratediscs.BDNFexpressionwassignificantlyincreasedininfiltrateda

6、nddegeneratesamples.SPandvascularendothelialgrowthfactor(VEGF)werehigherininfiltratedsamples.InvitrostimulationbyIL-1βinducedNGFinNPcells.Neurotropin-3wasinducedbytumournecrosisfactoralphainhumandermalmicrovascularendothelialcells(HDMECs).SPgeneandproteinexpressionwasincreasedinNPcel

7、lsbyIL-1β.CalcitoningenerelatedpeptidewasincreasedinSH-SY5Ycellsuponcytokinestimulation.VEGFwasinducedbyIL-1βandinterleukin-6inNPcells,whilstpleiotrophinwasdecreasedbyIL-1β.VEGFandpleiotrophinwereexpressedbySH-SY5Ycells,andVEGFbyHDMECs,butwerenotmodulatedbycytokines.Conclusions:There

8、leaseofcytok