Ginsenoside Rg1 promotes growth and extracellular matrix synthesis in degenerative human lumbar nucleus pulposus cells by inhibiting Wnt/β-catenin pathway
10.3969/j.issn.1000-4718.2018.04.020
- VernacularTitle:人参皂苷Rg1通过抑制Wnt/β-catenin通路促进退变人腰椎间盘髓核细胞的生长及胞外基质合成
- Author:
Hua LU
1
;
Lu YU
;
Huan-Huan ZHEN
;
Ru-Yin LIU
;
Zong-Jin YUE
Author Information
1. 河南中医药大学第二附属医院脊柱科
- Keywords:
Ginsenoside Rg1;
Human lumbar nucleus pulposus cells;
Cell viability;
Apoptosis;
Wnt/β-cate-nin pathway
- From:
Chinese Journal of Pathophysiology
2018;34(4):705-710
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To explore the role of ginsenoside Rg1 in the growth of degenerative human lumbar nucleus pulposus cells(HNPCs).METHODS:Cultured HNPCs were subjected to oxygen-glucose deprivation(OGD)to mimic the micro-environment of degenerative HNPCs.The morphological changes of the cells in control group and OGD group were observed under optical microscope.The cells were treated with ginsenoside Rg 1 at concentrations of 25,50 and 100 μmol/L.The expression of collagen II and aggrecan at mRNA and protein levels was determined by real -time PCR and Western blot analysis.The cell viability was measured by CCK-8 assay.The mRNA level of Ki67 was detected by real-time PCR.The apoptosis was analyzed by flow cytometry.The activity of caspase-3 was measured by a caspase-3 kit.The ex-pression of Wnt/β-catenin pathway-related proteins was determined by Western blot.Furthermore,the expression of Wnt/β-catenin pathway-related proteins,the cell viability and apoptosis,and the expression of extracellular matrix synthesis pro-teins were assessed after the cells were co-treated with LiCl and 100 μmol/L ginsenoside Rg1.RESULTS:Normal HNPCs attached on the cell culture plate faster, and were almost round with rich cytoplasm.However, the cell adherence was slower,and the cells were long fusiform with decreased cytoplasm after OGD treatment,indicating that the model of degen-erative HNPCs was successfully established.Compared with normal HNPCs,the expression of collagen II and aggrecan at mRNA and protein levels was decreased in OGD group(P<0.05),which was then increased after the cells were treated with ginsenoside Rg1 at 25,50 and 100 μmol/L(P<0.05).Compared with normal HNPCs,the cell viability and Ki67 expression were decreased in OGD group(P<0.05), which were increased after treatment with ginsenoside Rg 1(P<0.05).Meanwhile,the apoptotic rate and caspase-3 activity were significantly increased in OGD-treated cells(P<0.05), which were decreased after treatment with ginsenoside Rg 1(P<0.05).In addition,the activation of Wnt/β-catenin path-way was also inhibited by ginsenoside Rg 1 treatment at dose of 100 μmol/L(P<0.05).LiCl,a Wnt/β-catenin pathway agonist,obviously decreased the protective effects of ginenoside Rg 1 on OGD-induced cells(P<0.05),indicating that the Wnt/β-catenin pathway was involved in the protective effects of ginenoside Rg 1 on degenerative HNPCs.CONCLUSION:Ginsenoside Rg1 promotes growth and extracellular matrix synthesis of degenerative HNPCs through inhibiting Wnt /β-cate-nin pathway.This study will provide a new idea for prevention and treatment of degenerative HNPCs.
