title: Periodic mechanical stress induces the extracellular matrix expression and migration of rat nucleus pulposus cells by upregulating the expression of intergrin α1 and phosphorylation of downstream phospholipase Cγ1.
abstract: Intervertebral disk degeneration (IDD) is a major cause of low back pain and an important socioeconomic burden. Degradation of the extracellular matrix (ECM) of nucleus pulposus (NP) cells in the interverterbal disk is important for IDD. Stress of a suitable frequency and amplitude promotes the synthesis of the ECM of NP cells, however, the associated mechanisms remain to be fully elucidated The present study aimed to investigate the effect of integrin α1 on the migration and ECM synthesis of NP cells under soft periodic mechanical stress. Rat NP cells were isolated and plated onto slides, and were then treated with or without the use of a periodic mechanical stress system. The expression levels of integrin α1, α5 and αv, ECM collagen 2A1 (Col2A1) and aggrecan, and the phosphorylation of phospholipase C‑γ1 (PLCγ1) were measured using reverse transcription‑quantitative polymerase chain reaction and western blot analyses. Cell migration was assayed using a scratch experiment. The results showed that exposure to periodic mechanical stress significantly induced the mRNA expression levels of Col2A1 and aggrecan, cell migration, mRNA expression of integrin α1 and phosphorylation of PLC‑γ1 of the NP, compared with the control (P<0.05). Inhibition of the PLCγ1 protein by U73122 significantly decreased the ECM expression under periodic mechanical stress (P<0.05). Small interfering RNA‑mediated integrin α1 gene knockdown suppressed the mRNA expression levels of Col2A1 and aggrecan, and suppressed the migration and phosphorylation of PLCγ1 of the NP cells under periodic mechanical stress, compared with the control (P<0.05). In conclusion, periodic mechanical stress induced ECM expression and the migration of NP cells via upregulating the expression of integrin α1 and the phosphorylation of downstream PLCγ1. These findings provide novel information to aid the understanding of the pathogenesis and development of IDD.
authors.affiliation: Department of Orthopedics, Changzhou Second Hospital Affiliated to Nanjing Medical University, Changzhou, Jiangsu 213003, P.R. China.
authors.forename: Gongming
authors.initials: G
authors.lastname: Gao
authors.affiliation: Department of Orthopedics, Jintan People's Hospital Affiliated to Jiangsu University, Jintan, Jiangsu 213200, P.R. China.
authors.forename: Jin
authors.initials: J
authors.lastname: He
authors.affiliation: Department of Orthopedics, Changzhou Second Hospital Affiliated to Nanjing Medical University, Changzhou, Jiangsu 213003, P.R. China.
authors.forename: Luming
authors.initials: L
authors.lastname: Nong
authors.affiliation: Department of Orthopedics, Jintan People's Hospital Affiliated to Jiangsu University, Jintan, Jiangsu 213200, P.R. China.
authors.forename: Hua
authors.initials: H
authors.lastname: Xie
authors.affiliation: Department of Orthopedics, Changzhou Second Hospital Affiliated to Nanjing Medical University, Changzhou, Jiangsu 213003, P.R. China.
authors.forename: Yongjing
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Department of Orthopedics, Changzhou Second Hospital Affiliated to Nanjing Medical University, Changzhou, Jiangsu 213003, P.R. China.
authors.forename: Nanwei
authors.initials: N
authors.lastname: Xu
authors.affiliation: Department of Orthopedics, Changzhou Second Hospital Affiliated to Nanjing Medical University, Changzhou, Jiangsu 213003, P.R. China.
authors.forename: Dong
authors.initials: D
authors.lastname: Zhou
mesh_terms: D000818:Animals; D002465:Cell Movement; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D015870:Gene Expression; D039423:Integrin alpha1 / Q000235:genetics / Q000378:metabolism*; D055959:Intervertebral Disc Degeneration / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008297:Male; D000070614:Nucleus Pulposus / Q000166:cytology* / Q000378:metabolism*; D051966:Phospholipase C gamma / Q000378:metabolism*; D010766:Phosphorylation; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D013314:Stress, Mechanical
chemical_list: D024043:Collagen Type II; D039423:Integrin alpha1; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051966:Phospholipase C gamma; C000607812:phospholipase Cgamma1, rat