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[NF-κB signaling pathway regulate endplate chondrocytes in rat vitro natural degeneration model].
Zhonghua Yi Xue Za Zhi 2016; 96(27):2182-6ZY

Abstract

OBJECTIVE

To explore the relationship between nuclear factor (NF)-κB and vitro nature degeneration model of endplate chondrocyte in rats.

METHODS

Rats endplate chondrocytes were isolated and cultured in vitro.Rebulated vitro natural degeneration model and cells were divided into control group (P2 cells group), naturally passaged group (P5 cells group) and NF-κB signaling pathway inhibition group (added Bay11-7082 when the passaged to P5 cells). The changes of cellular morphology were observed by inverted phase contrast microscope, the phenotype of endplate chondrocyte were identified by toluidine blue staining, electromagnetic compatibility (EMC) were observed by alcian blue staining.Type Ⅱ collagen, sry related HMG box (SOX)-9, matrix metalloproteinase(MMP)13 and aggrecan genes were detected by real time-polymerase chain reaction (RT-PCR) to verify the degeneration mode.NF-κB transcriptional activity was assessed by examining cytosolic phosphorylated IκBα and nuclear phosphorylated p65 levels by Western blot.

RESULTS

With chondrocytes passing, the cells lost the original morphology gradually.Alcian blue stains observed EMC decreased in naturally passaged group.The leave of Type Ⅱ collagen (P5/P2=0.182, P<0.01), aggrecan (P5/P2=0.287, P<0.01) and SOX-9 (P5/P2=0.488, P<0.01) were significantly reduced, MMP13 (P5/P2=1.324, P<0.05) significantly increased.Western blot analysis showed that in P5 cells nuclear phosphorylation p65 and cytosolic levels of phosphorylated IκBα increased and Bay11-7082 treatment attenuated increase in nuclear phosphorylation p65 and blocked acidinduced increase in cytosolic levels of phosphorylated IκBα.Moreover, the leave of Type Ⅱ collagen (Bay11-7082/P5=4.173, P<0.01), aggrecan (Bay11-7082/P5=2.732, P<0.05) and SOX-9 (Bay11-7082/P5=1.567, P<0.05) significantlyincreased, MMP13 (Bay11-7082/P5=0.611, P<0.05) significantly reduced in treatment group.

CONCLUSION

Inhibitor NF-κB signaling pathway plays an important role in the vitro degeneration of endplate cartilage.Reasonable regulation of NF-κB signaling pathway may be a new way to prevent Intervertebral disc degeneration.

Authors+Show Affiliations

Department of Orthopedic Surgery, Yijishan Hosptial, Wannan Medical College, Wuhu 241001, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

chi

PubMed ID

27464547

Citation

Gao, Z, et al. "[NF-κB Signaling Pathway Regulate Endplate Chondrocytes in Rat Vitro Natural Degeneration Model]." Zhonghua Yi Xue Za Zhi, vol. 96, no. 27, 2016, pp. 2182-6.
Gao Z, Xu HG, Zhang XL, et al. [NF-κB signaling pathway regulate endplate chondrocytes in rat vitro natural degeneration model]. Zhonghua Yi Xue Za Zhi. 2016;96(27):2182-6.
Gao, Z., Xu, H. G., Zhang, X. L., Wang, H., Ma, M. M., Xiao, L., & Liu, X. (2016). [NF-κB signaling pathway regulate endplate chondrocytes in rat vitro natural degeneration model]. Zhonghua Yi Xue Za Zhi, 96(27), pp. 2182-6. doi:10.3760/cma.j.issn.0376-2491.2016.27.016.
Gao Z, et al. [NF-κB Signaling Pathway Regulate Endplate Chondrocytes in Rat Vitro Natural Degeneration Model]. Zhonghua Yi Xue Za Zhi. 2016 Jul 19;96(27):2182-6. PubMed PMID: 27464547.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [NF-κB signaling pathway regulate endplate chondrocytes in rat vitro natural degeneration model]. AU - Gao,Z, AU - Xu,H G, AU - Zhang,X L, AU - Wang,H, AU - Ma,M M, AU - Xiao,L, AU - Liu,X, PY - 2016/7/29/entrez PY - 2016/7/29/pubmed PY - 2017/3/25/medline SP - 2182 EP - 6 JF - Zhonghua yi xue za zhi JO - Zhonghua Yi Xue Za Zhi VL - 96 IS - 27 N2 - OBJECTIVE: To explore the relationship between nuclear factor (NF)-κB and vitro nature degeneration model of endplate chondrocyte in rats. METHODS: Rats endplate chondrocytes were isolated and cultured in vitro.Rebulated vitro natural degeneration model and cells were divided into control group (P2 cells group), naturally passaged group (P5 cells group) and NF-κB signaling pathway inhibition group (added Bay11-7082 when the passaged to P5 cells). The changes of cellular morphology were observed by inverted phase contrast microscope, the phenotype of endplate chondrocyte were identified by toluidine blue staining, electromagnetic compatibility (EMC) were observed by alcian blue staining.Type Ⅱ collagen, sry related HMG box (SOX)-9, matrix metalloproteinase(MMP)13 and aggrecan genes were detected by real time-polymerase chain reaction (RT-PCR) to verify the degeneration mode.NF-κB transcriptional activity was assessed by examining cytosolic phosphorylated IκBα and nuclear phosphorylated p65 levels by Western blot. RESULTS: With chondrocytes passing, the cells lost the original morphology gradually.Alcian blue stains observed EMC decreased in naturally passaged group.The leave of Type Ⅱ collagen (P5/P2=0.182, P<0.01), aggrecan (P5/P2=0.287, P<0.01) and SOX-9 (P5/P2=0.488, P<0.01) were significantly reduced, MMP13 (P5/P2=1.324, P<0.05) significantly increased.Western blot analysis showed that in P5 cells nuclear phosphorylation p65 and cytosolic levels of phosphorylated IκBα increased and Bay11-7082 treatment attenuated increase in nuclear phosphorylation p65 and blocked acidinduced increase in cytosolic levels of phosphorylated IκBα.Moreover, the leave of Type Ⅱ collagen (Bay11-7082/P5=4.173, P<0.01), aggrecan (Bay11-7082/P5=2.732, P<0.05) and SOX-9 (Bay11-7082/P5=1.567, P<0.05) significantlyincreased, MMP13 (Bay11-7082/P5=0.611, P<0.05) significantly reduced in treatment group. CONCLUSION: Inhibitor NF-κB signaling pathway plays an important role in the vitro degeneration of endplate cartilage.Reasonable regulation of NF-κB signaling pathway may be a new way to prevent Intervertebral disc degeneration. SN - 0376-2491 UR - https://www.unboundmedicine.com/medline/citation/27464547/[NF_κB_signaling_pathway_regulate_endplate_chondrocytes_in_rat_vitro_natural_degeneration_model]_ L2 - http://journal.yiigle.com/LinkIn.do?linkin_type=pubmed&amp;issn=0376-2491&amp;year=2016&amp;vol=96&amp;issue=27&amp;fpage=2182 DB - PRIME DP - Unbound Medicine ER -