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Published in: BMC Complementary Medicine and Therapies 1/2016

Open Access 01-12-2016 | Research article

Effects of panax notoginseng saponins on the osteogenic differentiation of rabbit bone mesenchymal stem cells through TGF-β1 signaling pathway

Authors: Yan Wang, Xuanping Huang, Yiyao Tang, Haiyun Lin, Nuo Zhou

Published in: BMC Complementary Medicine and Therapies | Issue 1/2016

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Abstract

Background

Panax Notoginseng is a well-known Chinese medicinal herb which has been used in China for treatment of bone fracture for hundreds of years. However, the specific biological mechanisms of osteogenic effect of PNS are not well understood.

Methods

In this study, newborn rabbit BMSCs were isolated, and then identified by the positive expression rates of cell surface markers, including CD29, CD45 and HLA-DR, which were detected by flow cytometry(FCM). After the lentivirus-induced cell model of TGF-β1 gene silencing was established, the interference efficiency was tested by q-PCR and Western blot, and the growth curve of silencing cells was drawn by MTT so as to grasp the growth rhythm of silencing cells. In the alizarin red-staining experiment, the effect of 100 mg/L PNS on the activity of intracellular ALP of TGF-β1 gene silencing BMSCs was detected, so as to observe the effect of 100 mg/L PNS on the formation of calcium nodes of gene silencing BMSCs.

Results

By separating rabbit BMSCs, the lentivirus-induced cell model of TGF-β1 gene silencing was established. Both TGF-β1 mRNA and protein expression were restrained significantly, and the target gene kept silence stably via the verification of q-PCR and Western blot; there was no significant differences of the growth curve between RNAi cells and normal cells; the activity of intracellular APL in 100 mg RNAi group was obviously lower than that in 100 mg group (p < 0.05), but higher than that in the normal group; in the alizarin red-staining experiment, it focused on the effects of PNS on the formation of calcium nodes of gene silencing BMSCs, which showed that calcium nodes could be formed in 100 mg RNAi group but its quantity was lower than that of 100 mg group (p < 0.05).

Conclusions

It was shown that silencing TGF-β1 gene could interrupt the osteogenic effects of PNS. PNS may have a promoting effect on osteogenic differentiation of rabbits’ BMSCs in vitro by up-regulating the gene expression of TGF-β1.
Literature
1.
go back to reference Gao XM. Chinese pharmacy. Beijing: China Press of Traditional Chinese Medicine; 2007. p. 296–7. Gao XM. Chinese pharmacy. Beijing: China Press of Traditional Chinese Medicine; 2007. p. 296–7.
2.
go back to reference Dong TT, Cui XM, Song ZH, Zhao KJ, Ji ZN, Lo CK, Tsim KW. Chemical assessment of roots of Panax notoginseng in China: regional and seasonal variations in its active constituents. J Agric Food Chem. 2003;51:4617–23.CrossRefPubMed Dong TT, Cui XM, Song ZH, Zhao KJ, Ji ZN, Lo CK, Tsim KW. Chemical assessment of roots of Panax notoginseng in China: regional and seasonal variations in its active constituents. J Agric Food Chem. 2003;51:4617–23.CrossRefPubMed
3.
go back to reference Uzayisenga R, Ayeka PA, Wang Y. Anti-diabetic potential of Panax notoginseng saponins (PNS): a review. Phytother Res. 2014;28:510–6.CrossRefPubMed Uzayisenga R, Ayeka PA, Wang Y. Anti-diabetic potential of Panax notoginseng saponins (PNS): a review. Phytother Res. 2014;28:510–6.CrossRefPubMed
4.
go back to reference Yang X, Xiong X, Wang J. Sanqi panax notoginseng injection for angina pectoris. Evid Based Complement Alternat Med. 2014;2014:963208.PubMedPubMedCentral Yang X, Xiong X, Wang J. Sanqi panax notoginseng injection for angina pectoris. Evid Based Complement Alternat Med. 2014;2014:963208.PubMedPubMedCentral
5.
go back to reference Liu N, Shan D, Li Y, Chen H, Gao Y, Huang Y. Panax notoginseng saponins attenuate phenotype switching of vascular smooth muscle cells induced by Notch3 silencing. Evid Based Complement Alternat Med. 2015;2015:162145.PubMedPubMedCentral Liu N, Shan D, Li Y, Chen H, Gao Y, Huang Y. Panax notoginseng saponins attenuate phenotype switching of vascular smooth muscle cells induced by Notch3 silencing. Evid Based Complement Alternat Med. 2015;2015:162145.PubMedPubMedCentral
6.
go back to reference Wen XD, Wang CZ, Yu C, Zhao L, Zhang Z, Matin A, Wang Y, Li P, Xiao SY, Du W, He TC, Yuan CS. Panax notoginseng attenuates experimental colitis in the azoxymethane/dextran sulfate sodium mouse model. Phytother Res. 2014;28:892–8.CrossRefPubMed Wen XD, Wang CZ, Yu C, Zhao L, Zhang Z, Matin A, Wang Y, Li P, Xiao SY, Du W, He TC, Yuan CS. Panax notoginseng attenuates experimental colitis in the azoxymethane/dextran sulfate sodium mouse model. Phytother Res. 2014;28:892–8.CrossRefPubMed
7.
go back to reference Yang J, Ren T, Li XD, Qiao ZG. The experiment study of total saponins of Panax notoginseng on promoting fracture healing. J Med Res. 2010;39:69–71. Yang J, Ren T, Li XD, Qiao ZG. The experiment study of total saponins of Panax notoginseng on promoting fracture healing. J Med Res. 2010;39:69–71.
8.
go back to reference Li XD, Wang JS, Chang B, Chen B, Guo C, Hou GQ, Huang DY, Du SX. Panax notoginseng saponins promotes proliferation and osteogenic differentiation of rat bone marrow stromal cells. J Ethnopharmacol. 2011;134:268–74.CrossRefPubMed Li XD, Wang JS, Chang B, Chen B, Guo C, Hou GQ, Huang DY, Du SX. Panax notoginseng saponins promotes proliferation and osteogenic differentiation of rat bone marrow stromal cells. J Ethnopharmacol. 2011;134:268–74.CrossRefPubMed
9.
go back to reference Liu Y, Hao F, Zhang H, Cao D, Lu X, Li X. Panax notoginseng saponins promote endothelial progenitor cell mobilization and attenuate atherosclerotic lesions in apolipoprotein E knockout mice. Cell Physiol Biochem. 2013;32:814–26.CrossRefPubMed Liu Y, Hao F, Zhang H, Cao D, Lu X, Li X. Panax notoginseng saponins promote endothelial progenitor cell mobilization and attenuate atherosclerotic lesions in apolipoprotein E knockout mice. Cell Physiol Biochem. 2013;32:814–26.CrossRefPubMed
10.
go back to reference Du WX, Duan SF, Yu XL, Yin LM. Panax notoginseng saponins suppress radiation-induced osteoporosis by regulating bone formation and resorption. Phytomedicine. 2015;22:813–19.CrossRef Du WX, Duan SF, Yu XL, Yin LM. Panax notoginseng saponins suppress radiation-induced osteoporosis by regulating bone formation and resorption. Phytomedicine. 2015;22:813–19.CrossRef
11.
go back to reference Bianco P, Rininucci M, Gronthos S, Robey PG. Bone marrow stromal stem cells: nature, biology, and potential applications. Stem Cells. 2001;19:180–92.CrossRefPubMed Bianco P, Rininucci M, Gronthos S, Robey PG. Bone marrow stromal stem cells: nature, biology, and potential applications. Stem Cells. 2001;19:180–92.CrossRefPubMed
12.
go back to reference Matsumura N, Ochi K, Ichimura M, Mizushima T, Harada H, Harada M. Study on free radicals and panceaticibrosis, pancereatic fibrosis induced by repeated injections of superoxide dismutase inhibitor. Oanceras. 2001;22:53–7. Matsumura N, Ochi K, Ichimura M, Mizushima T, Harada H, Harada M. Study on free radicals and panceaticibrosis, pancereatic fibrosis induced by repeated injections of superoxide dismutase inhibitor. Oanceras. 2001;22:53–7.
13.
go back to reference Ma L, Liu DJ, Li DT, Liu PT. Effects of different separate methods and culture conditions on growth proliferation and biological characteristics of rabbit bone marrow mesenchymal stem cells. J Clin Rehabil Tissue Eng Res. 2008;38:7401–06. Ma L, Liu DJ, Li DT, Liu PT. Effects of different separate methods and culture conditions on growth proliferation and biological characteristics of rabbit bone marrow mesenchymal stem cells. J Clin Rehabil Tissue Eng Res. 2008;38:7401–06.
14.
go back to reference Li X, Zhang Y, Qi G. Evaluation of isolation methods and culture conditions for rat bone marrow mesenchymal stem cells. Cytotechnology. 2013;65:323–34.CrossRefPubMed Li X, Zhang Y, Qi G. Evaluation of isolation methods and culture conditions for rat bone marrow mesenchymal stem cells. Cytotechnology. 2013;65:323–34.CrossRefPubMed
15.
go back to reference Friedenstein AJ, Chailakhyan RK, Gerasimov UV. Bone marrow osteogenic stem cells: in vitro cultivation and transplantation in diffusion chambers. Cell T issue Kinet. 1987;20:263–67. Friedenstein AJ, Chailakhyan RK, Gerasimov UV. Bone marrow osteogenic stem cells: in vitro cultivation and transplantation in diffusion chambers. Cell T issue Kinet. 1987;20:263–67.
16.
go back to reference Wang XF, Tong PJ, Jin HT. Approaches to isolate, culture and identify the rabbit marrow mesenchymal stem cells. J Tradit Chinese Orthop Traum. 2011;23:3–6. Wang XF, Tong PJ, Jin HT. Approaches to isolate, culture and identify the rabbit marrow mesenchymal stem cells. J Tradit Chinese Orthop Traum. 2011;23:3–6.
17.
go back to reference Liu Q, Shi Y, Wang H. Experimental study on isolation, culture and identify of rabbit bone marrow-derived mesenchymal stem cells in vitro. J Tissue Eng Reconstr Surg. 2013;17:1366–69. Liu Q, Shi Y, Wang H. Experimental study on isolation, culture and identify of rabbit bone marrow-derived mesenchymal stem cells in vitro. J Tissue Eng Reconstr Surg. 2013;17:1366–69.
19.
go back to reference Schuldiner M, Yanuka O, Itskovitz-Eldor J, Melton DA, Benvenisty N. Effects of eight growth factors on the differentiation of cells derived from human embryonic stem cells. Proc Natl Acad Sci U S A. 2000;97:11307–12.CrossRefPubMedPubMedCentral Schuldiner M, Yanuka O, Itskovitz-Eldor J, Melton DA, Benvenisty N. Effects of eight growth factors on the differentiation of cells derived from human embryonic stem cells. Proc Natl Acad Sci U S A. 2000;97:11307–12.CrossRefPubMedPubMedCentral
20.
go back to reference Wang DW, Pan H, Li HB, Wang HX, Chen YP. Influence of total saponins of Panax Notoginseng on the alcohol-induced differentiation of bone marrow stromal stem cells. J Clin Rehabil Tissue Eng Res. 2008;12:1410–13. Wang DW, Pan H, Li HB, Wang HX, Chen YP. Influence of total saponins of Panax Notoginseng on the alcohol-induced differentiation of bone marrow stromal stem cells. J Clin Rehabil Tissue Eng Res. 2008;12:1410–13.
21.
go back to reference Yao XM, Cheng Y, Fang F. Experimental study on YIGUTANG containing serum on osteoblast proliferation and ALP expression. J Zhejiang University Tradit Chinese Med. 2007;31:1–2. Yao XM, Cheng Y, Fang F. Experimental study on YIGUTANG containing serum on osteoblast proliferation and ALP expression. J Zhejiang University Tradit Chinese Med. 2007;31:1–2.
22.
go back to reference Maniatopoulos C, Sodek J, Melcher AH. Bone formation in vitro by stromal cells obtained from bone marrow of young adult rats. Cell Tissue Res. 1998;254:317–25. Maniatopoulos C, Sodek J, Melcher AH. Bone formation in vitro by stromal cells obtained from bone marrow of young adult rats. Cell Tissue Res. 1998;254:317–25.
23.
go back to reference Hilton MJ, Tu X, Wu X, Bai S, Zhao H, Kobayashi T, Kronenberg HM, Teitelbaum SL, Ross FP, Kopan R, Long F. Notch signaling maintains bone marrow mesenchymal progenitors by suppressing osteoblast differentiation. Nat Med. 2008;14:306–14.CrossRefPubMedPubMedCentral Hilton MJ, Tu X, Wu X, Bai S, Zhao H, Kobayashi T, Kronenberg HM, Teitelbaum SL, Ross FP, Kopan R, Long F. Notch signaling maintains bone marrow mesenchymal progenitors by suppressing osteoblast differentiation. Nat Med. 2008;14:306–14.CrossRefPubMedPubMedCentral
24.
go back to reference Dishowitz MI, Mutyaba PL, Takacs JD, Barr AM, Engiles JB, Ahn J, Hankenson KD. Systemic inhibition of canonical Notch signaling results in sustained callus inflammation and alters multiple phases of fracture healing. PLoS One. 2013;8:e68726.CrossRefPubMedPubMedCentral Dishowitz MI, Mutyaba PL, Takacs JD, Barr AM, Engiles JB, Ahn J, Hankenson KD. Systemic inhibition of canonical Notch signaling results in sustained callus inflammation and alters multiple phases of fracture healing. PLoS One. 2013;8:e68726.CrossRefPubMedPubMedCentral
25.
26.
go back to reference Chen B, Li XD, Liu DX, Wang H, Xie P, Liu ZY, Hou GQ, Chang B, Du SX. Canonical Wnt signaling is required for Panax Notoginseng Saponin-mediated attenuation of the RANKL/OPG ratio in bone marrow stromal cells during osteogenic differentiation. Phytomedicine. 2012;19:1029–34.CrossRefPubMed Chen B, Li XD, Liu DX, Wang H, Xie P, Liu ZY, Hou GQ, Chang B, Du SX. Canonical Wnt signaling is required for Panax Notoginseng Saponin-mediated attenuation of the RANKL/OPG ratio in bone marrow stromal cells during osteogenic differentiation. Phytomedicine. 2012;19:1029–34.CrossRefPubMed
27.
go back to reference Plaisant M, Fontaine C, Cousin W, Rochet N, Dani C, Peraldi P. Activation of Hedgehog signaling inhibits osteoblast differentiation of human mesenchymal stem cells. Stem Cells. 2009;27:703–13.CrossRefPubMed Plaisant M, Fontaine C, Cousin W, Rochet N, Dani C, Peraldi P. Activation of Hedgehog signaling inhibits osteoblast differentiation of human mesenchymal stem cells. Stem Cells. 2009;27:703–13.CrossRefPubMed
28.
go back to reference Hu Y, Chan E, Wang SX, Li B. Activation of p38 mitogen-activated protein kinase is required for osteoblast differentiation. Endocrinology. 2003;114:2068–74.CrossRef Hu Y, Chan E, Wang SX, Li B. Activation of p38 mitogen-activated protein kinase is required for osteoblast differentiation. Endocrinology. 2003;114:2068–74.CrossRef
Metadata
Title
Effects of panax notoginseng saponins on the osteogenic differentiation of rabbit bone mesenchymal stem cells through TGF-β1 signaling pathway
Authors
Yan Wang
Xuanping Huang
Yiyao Tang
Haiyun Lin
Nuo Zhou
Publication date
01-12-2016
Publisher
BioMed Central
Published in
BMC Complementary Medicine and Therapies / Issue 1/2016
Electronic ISSN: 2662-7671
DOI
https://doi.org/10.1186/s12906-016-1304-9

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