Transient receptor potential vanilloid 1-based gene therapy alleviates orthodontic pain in rats
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摘要
Orthodontic pain that is induced by tooth movement is an important sequela of orthodontic treatment and has a significant effect on patient quality of life. Studies have shown that the high expression of transient receptor potential vanilloid 1(TRPV1) in trigeminal ganglions plays a vital role in the transmission and modulation of orofacial pain. However, little is known about the role of TRPV1 in orthodontic pain. In this study, male Sprague–Dawley rats were randomly assigned to six groups to study the role of TRPV1 in the modulation of tooth-movement pain. The expression levels of TRPV1 mRNA and protein were determined by real-time PCR and western blot, respectively. Moreover, pain levels were assessed using the rat grimace scale(RGS). The role of TRPV1 in modulating tooth-movement pain was examined by injecting a TRPV1 antagonist into the trigeminal ganglia of rats. A lentivirus containing a TRPV1 shRNA sequence was constructed and transduced into the rats' trigeminal ganglia. The results showed that the expression levels of TRPV1 protein and m RNA were elevated following tooth-movement pain. Pain levels increased rapidly on the1 stday, peaked on the 3~(rd) day and returned to baseline on the 14 thday. The TRPV1 antagonist significantly reduced toothmovement pain. The lentivirus containing a TRPV1 shRNA sequence was able to inhibit the expression of TRPV1 and relieved toothmovement pain. In conclusion, TRPV1-based gene therapy may be a treatment strategy for the relief of orthodontic pain.
Orthodontic pain that is induced by tooth movement is an important sequela of orthodontic treatment and has a significant effect on patient quality of life. Studies have shown that the high expression of transient receptor potential vanilloid 1(TRPV1) in trigeminal ganglions plays a vital role in the transmission and modulation of orofacial pain. However, little is known about the role of TRPV1 in orthodontic pain. In this study, male Sprague–Dawley rats were randomly assigned to six groups to study the role of TRPV1 in the modulation of tooth-movement pain. The expression levels of TRPV1 mRNA and protein were determined by real-time PCR and western blot, respectively. Moreover, pain levels were assessed using the rat grimace scale(RGS). The role of TRPV1 in modulating tooth-movement pain was examined by injecting a TRPV1 antagonist into the trigeminal ganglia of rats. A lentivirus containing a TRPV1 shRNA sequence was constructed and transduced into the rats' trigeminal ganglia. The results showed that the expression levels of TRPV1 protein and m RNA were elevated following tooth-movement pain. Pain levels increased rapidly on the1 stday, peaked on the 3~(rd) day and returned to baseline on the 14 thday. The TRPV1 antagonist significantly reduced toothmovement pain. The lentivirus containing a TRPV1 shRNA sequence was able to inhibit the expression of TRPV1 and relieved toothmovement pain. In conclusion, TRPV1-based gene therapy may be a treatment strategy for the relief of orthodontic pain.
Orthodontic pain that is induced by tooth movement is an important sequela of orthodontic treatment and has a significant effect on patient quality of life. Studies have shown that the high expression of transient receptor potential vanilloid 1(TRPV1) in trigeminal ganglions plays a vital role in the transmission and modulation of orofacial pain. However, little is known about the role of TRPV1 in orthodontic pain. In this study, male Sprague–Dawley rats were randomly assigned to six groups to study the role of TRPV1 in the modulation of tooth-movement pain. The expression levels of TRPV1 mRNA and protein were determined by real-time PCR and western blot, respectively. Moreover, pain levels were assessed using the rat grimace scale(RGS). The role of TRPV1 in modulating tooth-movement pain was examined by injecting a TRPV1 antagonist into the trigeminal ganglia of rats. A lentivirus containing a TRPV1 shRNA sequence was constructed and transduced into the rats' trigeminal ganglia. The results showed that the expression levels of TRPV1 protein and m RNA were elevated following tooth-movement pain. Pain levels increased rapidly on the1 stday, peaked on the 3~(rd) day and returned to baseline on the 14 thday. The TRPV1 antagonist significantly reduced toothmovement pain. The lentivirus containing a TRPV1 shRNA sequence was able to inhibit the expression of TRPV1 and relieved toothmovement pain. In conclusion, TRPV1-based gene therapy may be a treatment strategy for the relief of orthodontic pain.
引文
1.Banerjee,S.,Banerjee,R.,Shenoy,U.,Agarkar,S.&Bhattacharya,S.Effect of orthodontic pain on quality of life of patients undergoing orthodontic treatment.Indian J.Dent.Res.29,4-9(2018).
2.Campos,M.J.,Fraga,M.R.,Raposo,N.R.,Ferreira,A.P.&Vitral,R.W.Assessment of pain experience in adults and children after bracket bonding and initial archwire insertion.Dent.Press J.Orthod.18,32-37(2013).
3.Rakhshan,H.&Rakhshan,V.Pain and discomfort perceived during the initial stage of active fixed orthodontic treatment.Saudi Dent.J.27,81-87(2015).
4.Krishnan,V.Orthodontic pain:from causes to management--a review.Eur.J.Orthod.29,170-179(2007).
5.Bergius,M.,Berggren,U.&Kiliaridis,S.Experience of pain during an orthodontic procedure.Eur.J.Oral.Sci.110,92-98(2002).
6.Long,H.et al.Current advances in orthodontic pain.Int.J.Oral.Sci.8,67-75(2016).
7.Gupta,M.et al.Controlling pain during orthodontic fixed appliance therapy with non-steroidal anti-inflammatory drugs(NSAID):a randomized,double-blinded,placebo-controlled study.J.Orofac.Orthop.75,471-476(2014).
8.Yadav,S.et al.Effect of low-frequency mechanical vibration on orthodontic tooth movement.Am.J.Orthod.Dentofac.Orthop.148,440-449(2015).
9.Deana,N.F.,Zaror,C.,Sandoval,P.&Alves,N.Effectiveness of low-level laser therapy in reducing orthodontic pain:a systematic review and meta-analysis.Pain.Res.Manag.2017,8560652(2017).
10.Sawada,A.,Usui,N.,Shimazaki,K.,Taira,M.&Ono,T.The effects of cognitive behavioral therapy on experimental orthodontic pain.Orthod.Waves 74,10-14(2015).
11.Prabhakar,A.R.,Paul,J.M.&Basappa,N.Gene therapy and its implications in dentistry.Int.J.Clin.Pediatr.Dent.4,85-92(2011).
12.Hussain,A.S.,Al Toubity,M.J.&Elias,W.Y.Methodologies in orthodontic pain management:a review.Open Dent.J.11,492-497(2017).
13.Glorioso,J.C.&Fink,D.J.Gene therapy for pain:introduction to the special issue.Gene Ther.16,453-454(2009).
14.Simonato,M.et al.Progress in gene therapy for neurological disorders.Nat.Rev.Neurol.9,277-291(2013).
15.Leung,R.K.&Whittaker,P.A.RNA interference:from gene silencing to genespecific therapeutics.Pharmacol.Ther.107,222-239(2005).
16.Rohl,T.&Kurreck,J.RNA interference in pain research.J.Neurochem.99,371-380(2006).
17.Zychowska,M.et al.Participation of pro-and anti-nociceptive interleukins in botulinum toxin A-induced analgesia in a rat model of neuropathic pain.Eur.J.Pharmacol.791,377-388(2016).
18.Singh,A.K.&Vinayak,M.Anti-nociceptive effect of resveratrol during inflammatory hyperalgesia via differential regulation of pro-inflammatory mediators.Phytother.Res.30,1164-1171(2016).
19.Gunthorpe,M.J.&Chizh,B.A.Clinical development of TRPV1 antagonists:targeting a pivotal point in the pain pathway.Drug Discov.Today 14,56-67(2009).
20.Caterina,M.J.et al.The capsaicin receptor:a heat-activated ion channel in the pain pathway.Nature 389,816-824(1997).
21.Zheng,J.Molecular mechanism of TRPchannels.Compr.Physiol.3,221-242(2013).
22.Ghilardi,J.R.et al.Selective blockade of the capsaicin receptor TRPV1 attenuates bone cancer pain.J.Neurosci.25,3126-3131(2005).
23.Honore,P.et al.A-425619[1-isoquinolin-5-yl-3-(4-trifluoromethyl-benzyl)-urea],a novel transient receptor potential type V1 receptor antagonist,relieves pathophysiological pain associated with inflammation and tissue injury in rats.J.Pharmacol.Exp.Ther.314,410-421(2005).
24.Qiao,H.,Gao,Y.,Zhang,C.&Zhou,H.Increased expression of TRPV1 in the trigeminal ganglion is involved in orofacial pain during experimental tooth movement in rats.Eur.J.Oral.Sci.123,17-23(2015).
25.Ohkura,M.et al.Orthodontic force application upregulated pain-associated prostaglandin-I2/PGI2-receptor/TRPV1 pathway-related gene expression in rat molars.Odontology 106,2-10(2018).
26.Gao,Y.et al.Blocking of TRPV-1 in the parodontium relieves orthodontic pain by inhibiting the expression of TRPV-1 in the trigeminal ganglion during experimental tooth movement in rats.Neurosci.Lett.628,67-72(2016).
27.Zhang,C.D.et al.Expression of TRPV1 and CGRP in rat trigeminal ganglion during orthodontic tooth movement.Shanghai Kou.Qiang.Yi.Xue.24,6-12(2015).
28.Gunthorpe,M.J.et al.Identification and characterisation of SB-366791,a potent and selective vanilloid receptor(VR1/TRPV1)antagonist.Neuropharmacology 46,133-149(2004).
29.Gavva,N.R.et al.AMG 9810[(E)-3-(4-t-butylphenyl)-N-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)acrylamide],a novel vanilloid receptor 1(TRPV1)antagonist with antihyperalgesic properties.J.Pharmacol.Exp.Ther.313,474-484(2005).
30.El Kouhen,R.et al.A-425619[1-isoquinolin-5-yl-3-(4-trifluoromethyl-benzyl)-urea],a novel and selective transient receptor potential type V1 receptor antagonist,blocks channel activation by vanilloids,heat,and acid.J.Pharmacol.Exp.Ther.314,400-409(2005).
31.Behrendt,H.J.,Germann,T.,Gillen,C.,Hatt,H.&Jostock,R.Characterization of the mouse cold-menthol receptor TRPM8 and vanilloid receptor type-1 VR1 using a fluorometric imaging plate reader(FLIPR)assay.Br.J.Pharmacol.141,737-745(2004).
32.Wiskur,B.J.et al.A novel Trpv1 receptor antagonist Jnj-17203212 attenuates colonic hypersensitivity in rats.Methods Find.Exp.Clin.Pharmacol.32,557-564(2010).
33.Teramoto,S.,Ishii,T.,Matsuse,T.&Fukuchi,Y.Recombinant adeno-associated virus vectors efficiently transduce foreign gene into bovine aortic endothelial cells:comparison with adenovirus vectors.Jpn.J.Pharmacol.84,206-212(2000).
34.Delenda,C.Lentiviral vectors:optimization of packaging,transduction and gene expression.J.Gene Med.6,S125-S138(2004).
35.Benavides,O.J.,Satlin,L.,Burrow,C.,Wilson,P.&Herold,B.Herpes simplex virus(HSV)as a model vector for gene therapy for renal disease.J.Am.Soc.Nephrol.13,125a(2002).
36.Daya,S.&Berns,K.I.Gene therapy using adeno-associated virus vectors.Clin.Microbiol.Rev.21,583-593(2008).
37.Ogawa,N.et al.Gene therapy for neuropathic pain by silencing of TNF-alpha expression with lentiviral vectors targeting the dorsal root ganglion in mice.PLoSONE 9,e92073(2014).
38.Wang,W.,Cao,X.H.,Liu,C.J.&Liu,L.J.Cannabinoid WIN 55,212-2 inhibits TRPV1in trigeminal ganglion neurons via PKA and PKC pathways.Neurol.Sci.33,79-85(2012).
39.Sanz-Salvador,L.,Andres-Borderia,A.,Ferrer-Montiel,A.&Planells-Cases,R.Agonist-and Ca2+-dependent desensitization of TRPV1 channel targets the receptor to lysosomes for degradation.J.Biol.Chem.287,19462-19471(2012).
40.Lappin,S.C.,Randall,A.D.,Gunthorpe,M.J.&Morisset,V.TRPV1 antagonist,SB-366791,inhibits glutamatergic synaptic transmission in rat spinal dorsal horn following peripheral inflammation.Eur.J.Pharmacol.540,73-81(2006).
41.Christoph,T.et al.Investigation of TRPV1 loss-of-function phenotypes in transgenic shRNA expressing and knockout mice.Mol.Cell.Neurosci.37,579-589(2008).
42.Nakamura,A.et al.G protein-gated inwardly rectifying potassium(KIR3)channels play a primary role in the antinociceptive effect of oxycodone,but not morphine,at supraspinal sites.Br.J.Pharmacol.171,253-264(2014).
43.Kilkenny,C.,Browne,W.J.,Cuthill,I.C.,Emerson,M.&Altman,D.G.Improving bioscience research reporting:the ARRIVE guidelines for reporting animal research.J.Pharmacol.Pharmacother.1,94-99(2010).
44.Liao,L.et al.Evaluation of pain in rats through facial expression following experimental tooth movement.Eur.J.Oral.Sci.122,121-124(2014).
2.Campos,M.J.,Fraga,M.R.,Raposo,N.R.,Ferreira,A.P.&Vitral,R.W.Assessment of pain experience in adults and children after bracket bonding and initial archwire insertion.Dent.Press J.Orthod.18,32-37(2013).
3.Rakhshan,H.&Rakhshan,V.Pain and discomfort perceived during the initial stage of active fixed orthodontic treatment.Saudi Dent.J.27,81-87(2015).
4.Krishnan,V.Orthodontic pain:from causes to management--a review.Eur.J.Orthod.29,170-179(2007).
5.Bergius,M.,Berggren,U.&Kiliaridis,S.Experience of pain during an orthodontic procedure.Eur.J.Oral.Sci.110,92-98(2002).
6.Long,H.et al.Current advances in orthodontic pain.Int.J.Oral.Sci.8,67-75(2016).
7.Gupta,M.et al.Controlling pain during orthodontic fixed appliance therapy with non-steroidal anti-inflammatory drugs(NSAID):a randomized,double-blinded,placebo-controlled study.J.Orofac.Orthop.75,471-476(2014).
8.Yadav,S.et al.Effect of low-frequency mechanical vibration on orthodontic tooth movement.Am.J.Orthod.Dentofac.Orthop.148,440-449(2015).
9.Deana,N.F.,Zaror,C.,Sandoval,P.&Alves,N.Effectiveness of low-level laser therapy in reducing orthodontic pain:a systematic review and meta-analysis.Pain.Res.Manag.2017,8560652(2017).
10.Sawada,A.,Usui,N.,Shimazaki,K.,Taira,M.&Ono,T.The effects of cognitive behavioral therapy on experimental orthodontic pain.Orthod.Waves 74,10-14(2015).
11.Prabhakar,A.R.,Paul,J.M.&Basappa,N.Gene therapy and its implications in dentistry.Int.J.Clin.Pediatr.Dent.4,85-92(2011).
12.Hussain,A.S.,Al Toubity,M.J.&Elias,W.Y.Methodologies in orthodontic pain management:a review.Open Dent.J.11,492-497(2017).
13.Glorioso,J.C.&Fink,D.J.Gene therapy for pain:introduction to the special issue.Gene Ther.16,453-454(2009).
14.Simonato,M.et al.Progress in gene therapy for neurological disorders.Nat.Rev.Neurol.9,277-291(2013).
15.Leung,R.K.&Whittaker,P.A.RNA interference:from gene silencing to genespecific therapeutics.Pharmacol.Ther.107,222-239(2005).
16.Rohl,T.&Kurreck,J.RNA interference in pain research.J.Neurochem.99,371-380(2006).
17.Zychowska,M.et al.Participation of pro-and anti-nociceptive interleukins in botulinum toxin A-induced analgesia in a rat model of neuropathic pain.Eur.J.Pharmacol.791,377-388(2016).
18.Singh,A.K.&Vinayak,M.Anti-nociceptive effect of resveratrol during inflammatory hyperalgesia via differential regulation of pro-inflammatory mediators.Phytother.Res.30,1164-1171(2016).
19.Gunthorpe,M.J.&Chizh,B.A.Clinical development of TRPV1 antagonists:targeting a pivotal point in the pain pathway.Drug Discov.Today 14,56-67(2009).
20.Caterina,M.J.et al.The capsaicin receptor:a heat-activated ion channel in the pain pathway.Nature 389,816-824(1997).
21.Zheng,J.Molecular mechanism of TRPchannels.Compr.Physiol.3,221-242(2013).
22.Ghilardi,J.R.et al.Selective blockade of the capsaicin receptor TRPV1 attenuates bone cancer pain.J.Neurosci.25,3126-3131(2005).
23.Honore,P.et al.A-425619[1-isoquinolin-5-yl-3-(4-trifluoromethyl-benzyl)-urea],a novel transient receptor potential type V1 receptor antagonist,relieves pathophysiological pain associated with inflammation and tissue injury in rats.J.Pharmacol.Exp.Ther.314,410-421(2005).
24.Qiao,H.,Gao,Y.,Zhang,C.&Zhou,H.Increased expression of TRPV1 in the trigeminal ganglion is involved in orofacial pain during experimental tooth movement in rats.Eur.J.Oral.Sci.123,17-23(2015).
25.Ohkura,M.et al.Orthodontic force application upregulated pain-associated prostaglandin-I2/PGI2-receptor/TRPV1 pathway-related gene expression in rat molars.Odontology 106,2-10(2018).
26.Gao,Y.et al.Blocking of TRPV-1 in the parodontium relieves orthodontic pain by inhibiting the expression of TRPV-1 in the trigeminal ganglion during experimental tooth movement in rats.Neurosci.Lett.628,67-72(2016).
27.Zhang,C.D.et al.Expression of TRPV1 and CGRP in rat trigeminal ganglion during orthodontic tooth movement.Shanghai Kou.Qiang.Yi.Xue.24,6-12(2015).
28.Gunthorpe,M.J.et al.Identification and characterisation of SB-366791,a potent and selective vanilloid receptor(VR1/TRPV1)antagonist.Neuropharmacology 46,133-149(2004).
29.Gavva,N.R.et al.AMG 9810[(E)-3-(4-t-butylphenyl)-N-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)acrylamide],a novel vanilloid receptor 1(TRPV1)antagonist with antihyperalgesic properties.J.Pharmacol.Exp.Ther.313,474-484(2005).
30.El Kouhen,R.et al.A-425619[1-isoquinolin-5-yl-3-(4-trifluoromethyl-benzyl)-urea],a novel and selective transient receptor potential type V1 receptor antagonist,blocks channel activation by vanilloids,heat,and acid.J.Pharmacol.Exp.Ther.314,400-409(2005).
31.Behrendt,H.J.,Germann,T.,Gillen,C.,Hatt,H.&Jostock,R.Characterization of the mouse cold-menthol receptor TRPM8 and vanilloid receptor type-1 VR1 using a fluorometric imaging plate reader(FLIPR)assay.Br.J.Pharmacol.141,737-745(2004).
32.Wiskur,B.J.et al.A novel Trpv1 receptor antagonist Jnj-17203212 attenuates colonic hypersensitivity in rats.Methods Find.Exp.Clin.Pharmacol.32,557-564(2010).
33.Teramoto,S.,Ishii,T.,Matsuse,T.&Fukuchi,Y.Recombinant adeno-associated virus vectors efficiently transduce foreign gene into bovine aortic endothelial cells:comparison with adenovirus vectors.Jpn.J.Pharmacol.84,206-212(2000).
34.Delenda,C.Lentiviral vectors:optimization of packaging,transduction and gene expression.J.Gene Med.6,S125-S138(2004).
35.Benavides,O.J.,Satlin,L.,Burrow,C.,Wilson,P.&Herold,B.Herpes simplex virus(HSV)as a model vector for gene therapy for renal disease.J.Am.Soc.Nephrol.13,125a(2002).
36.Daya,S.&Berns,K.I.Gene therapy using adeno-associated virus vectors.Clin.Microbiol.Rev.21,583-593(2008).
37.Ogawa,N.et al.Gene therapy for neuropathic pain by silencing of TNF-alpha expression with lentiviral vectors targeting the dorsal root ganglion in mice.PLoSONE 9,e92073(2014).
38.Wang,W.,Cao,X.H.,Liu,C.J.&Liu,L.J.Cannabinoid WIN 55,212-2 inhibits TRPV1in trigeminal ganglion neurons via PKA and PKC pathways.Neurol.Sci.33,79-85(2012).
39.Sanz-Salvador,L.,Andres-Borderia,A.,Ferrer-Montiel,A.&Planells-Cases,R.Agonist-and Ca2+-dependent desensitization of TRPV1 channel targets the receptor to lysosomes for degradation.J.Biol.Chem.287,19462-19471(2012).
40.Lappin,S.C.,Randall,A.D.,Gunthorpe,M.J.&Morisset,V.TRPV1 antagonist,SB-366791,inhibits glutamatergic synaptic transmission in rat spinal dorsal horn following peripheral inflammation.Eur.J.Pharmacol.540,73-81(2006).
41.Christoph,T.et al.Investigation of TRPV1 loss-of-function phenotypes in transgenic shRNA expressing and knockout mice.Mol.Cell.Neurosci.37,579-589(2008).
42.Nakamura,A.et al.G protein-gated inwardly rectifying potassium(KIR3)channels play a primary role in the antinociceptive effect of oxycodone,but not morphine,at supraspinal sites.Br.J.Pharmacol.171,253-264(2014).
43.Kilkenny,C.,Browne,W.J.,Cuthill,I.C.,Emerson,M.&Altman,D.G.Improving bioscience research reporting:the ARRIVE guidelines for reporting animal research.J.Pharmacol.Pharmacother.1,94-99(2010).
44.Liao,L.et al.Evaluation of pain in rats through facial expression following experimental tooth movement.Eur.J.Oral.Sci.122,121-124(2014).