TY - JOUR
T1 - Ligustrazine Prevents Intervertebral Disc Degeneration via Suppression of Aberrant TGF β Activation in Nucleus Pulposus Cells
AU - Liu, Shufen
AU - Cheng, Yuhao
AU - Tan, Yuqi
AU - Dong, Jingcheng
AU - Bian, Qin
N1 - Funding Information:
β Activation in Nucleus Pulposus Cells Liu Shufen xiaoliufenfen@163.com 1 Cheng Yuhao ray800201@gmail.com 2 Tan Yuqi ytan19@jhmi.edu 2 https://orcid.org/0000-0002-5194-367X Dong Jingcheng jcdong2004@126.com 3 https://orcid.org/0000-0001-7283-0132 Bian Qin bianqin213@gmail.com 3 Rodgers William B. 1 Longhua Hospital Shanghai University of Traditional Chinese Medicine Shanghai China shutcm.edu.cn 2 Institute for Cell Engineering Johns Hopkins University School of Medicine Baltimore MD USA jhu.edu 3 The Institutes of Integrative Medicine Fudan University Shanghai China fudan.edu.cn 2019 2 12 2019 2019 07 06 2019 10 08 2019 23 08 2019 2 12 2019 2019 Copyright © 2019 Shufen Liu et al. This is an open access article distributed under the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Objectives . Aberrant transforming growth factor β (TGF β ) activation is detrimental to both nucleus pulposus (NP) cells and cartilage endplates (CEPs), which can lead to intervertebral disc degeneration (IDD). Ligustrazine (LIG) reduces the expression of inflammatory factors and TGF β 1 in hypertrophic CEP to prevent IDD. In this study, we investigate the effects of LIG on NP cells and the TGF β signaling. Design . LIG was injected to the lumbar spinal instability (LSI) mouse model. The effect of LIG was evaluated by intervertebral disc (IVD) score in the LSI mouse model. The expression of activated TGF β was examined using immunostaining with pSmad2/3 antibody. The upright posture (UP) rat model was also treated and evaluated in the same manner to assess the effect of LIG. In ex vivo study, IVDs from four-week old mice were isolated and treated with 10 −5 , 10 −6 , and 10 −7 M of LIG. We used western blot to detect activated TGF β expression. TGF β -treated human nucleus pulposus cells (HNPCs) were cotreated with optimized dose of LIG in vitro . Immunofluorescence staining was performed to determine pSmad2/3, connective tissue growth factor (CCN2), and aggrecan (ACAN) expression levels. Results . IVD score and the percentage of pSmad2/3+ NP cells were low in LIG-treated LSI mice in comparison with LSI mice, but close to the levels in the Sham group. Similarly, LIG reduced the overexpression of TGF β 1 in NP cells. The inhibitory effect of LIG was dose dependent. A dose of 10 −5 M LIG not only strongly attenuated Smad2/3 phosphorylation in TGF β -treated IVD ex vivo but also suppressed pSmad2/3, CCN2, and ACAN expression in TGF β -treated NP cells in vitro . Conclusions . LIG prevents IDD via suppression of TGF β overactivation in NP cells. National Natural Science Foundation of China 81573992 Development Project of Shanghai Peak Disciplines-Integrative Medicine 20150407
Publisher Copyright:
© 2019 Shufen Liu et al.
PY - 2019
Y1 - 2019
N2 - Objectives. Aberrant transforming growth factor β (TGFβ) activation is detrimental to both nucleus pulposus (NP) cells and cartilage endplates (CEPs), which can lead to intervertebral disc degeneration (IDD). Ligustrazine (LIG) reduces the expression of inflammatory factors and TGFβ1 in hypertrophic CEP to prevent IDD. In this study, we investigate the effects of LIG on NP cells and the TGFβ signaling. Design. LIG was injected to the lumbar spinal instability (LSI) mouse model. The effect of LIG was evaluated by intervertebral disc (IVD) score in the LSI mouse model. The expression of activated TGFβ was examined using immunostaining with pSmad2/3 antibody. The upright posture (UP) rat model was also treated and evaluated in the same manner to assess the effect of LIG. In ex vivo study, IVDs from four-week old mice were isolated and treated with 10-5, 10-6, and 10-7 M of LIG. We used western blot to detect activated TGFβ expression. TGFβ-treated human nucleus pulposus cells (HNPCs) were cotreated with optimized dose of LIG in vitro. Immunofluorescence staining was performed to determine pSmad2/3, connective tissue growth factor (CCN2), and aggrecan (ACAN) expression levels. Results. IVD score and the percentage of pSmad2/3+ NP cells were low in LIG-treated LSI mice in comparison with LSI mice, but close to the levels in the Sham group. Similarly, LIG reduced the overexpression of TGFβ1 in NP cells. The inhibitory effect of LIG was dose dependent. A dose of 10-5 M LIG not only strongly attenuated Smad2/3 phosphorylation in TGFβ-treated IVD ex vivo but also suppressed pSmad2/3, CCN2, and ACAN expression in TGFβ-treated NP cells in vitro. Conclusions. LIG prevents IDD via suppression of TGFβ overactivation in NP cells.
AB - Objectives. Aberrant transforming growth factor β (TGFβ) activation is detrimental to both nucleus pulposus (NP) cells and cartilage endplates (CEPs), which can lead to intervertebral disc degeneration (IDD). Ligustrazine (LIG) reduces the expression of inflammatory factors and TGFβ1 in hypertrophic CEP to prevent IDD. In this study, we investigate the effects of LIG on NP cells and the TGFβ signaling. Design. LIG was injected to the lumbar spinal instability (LSI) mouse model. The effect of LIG was evaluated by intervertebral disc (IVD) score in the LSI mouse model. The expression of activated TGFβ was examined using immunostaining with pSmad2/3 antibody. The upright posture (UP) rat model was also treated and evaluated in the same manner to assess the effect of LIG. In ex vivo study, IVDs from four-week old mice were isolated and treated with 10-5, 10-6, and 10-7 M of LIG. We used western blot to detect activated TGFβ expression. TGFβ-treated human nucleus pulposus cells (HNPCs) were cotreated with optimized dose of LIG in vitro. Immunofluorescence staining was performed to determine pSmad2/3, connective tissue growth factor (CCN2), and aggrecan (ACAN) expression levels. Results. IVD score and the percentage of pSmad2/3+ NP cells were low in LIG-treated LSI mice in comparison with LSI mice, but close to the levels in the Sham group. Similarly, LIG reduced the overexpression of TGFβ1 in NP cells. The inhibitory effect of LIG was dose dependent. A dose of 10-5 M LIG not only strongly attenuated Smad2/3 phosphorylation in TGFβ-treated IVD ex vivo but also suppressed pSmad2/3, CCN2, and ACAN expression in TGFβ-treated NP cells in vitro. Conclusions. LIG prevents IDD via suppression of TGFβ overactivation in NP cells.
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U2 - 10.1155/2019/5601734
DO - 10.1155/2019/5601734
M3 - Article
C2 - 31886227
AN - SCOPUS:85077003042
SN - 2314-6133
VL - 2019
JO - BioMed research international
JF - BioMed research international
M1 - 5601734
ER -