Tougu Xiaotong Granule containing serum induces cartilage differentiation of bone marrow stromal stem cells

doi:10.3969/j.issn.2095-4344.2013.32.018

Abstract

Abstract:

BACKGROUND: Bone marrow stromal stem cells have multilineage differentiation capacity and can differentiate into transparent chondrocytes under certain conditions, which can provide new thoughts for treatment of osteoarthritis.
OBJECTIVE: To investigate the effects of Tougu Xiaotong Granule containing serum on the cartilage differentiation of bone marrow stromal stem cells.
METHODS: Bone marrow stromal stem cells from Sprague-Dawley rat limbs were cultured in vitro, and those cells at passage 3 were used in the study. Cells were divided into six groups: saline serum group, Tougu Xiaotong Granule water extract group, Tougu Xiaotong Granule alcohol extract group, chondroinductive group, Tougu Xiaotong Granule water extract and chondroinductive group, Tougu Xiaotong Granule alcohol extract and chondroinductive group. The Sox9, collagen Ⅱ, and collagen X mRNA and protein expression levels were tested by real-time polymerase chain reaction and western blot analysis.
RESULTS AND CONCLUSION: After cells were intervened with drug-containing serum for 14 days, the Sox9, collagen Ⅱ, and collagen X mRNA and protein expression in Tougu Xiaotong Granule water extract group, Tougu Xiaotong Granule alcohol extract group, chondroinductive group, Tougu Xiaotong Granule water extract and chondroinductive group, Tougu Xiaotong Granule alcohol extract and chondroinductive group were significantly higher than that in saline serum group (P < 0.05, P < 0.01). In addition, the chondroinductive group, Tougu Xiaotong Granule water extract serum and chondroinductive group, Tougu Xiaotong Granule alcohol extract and chondroinductive group showed significantly higher expression levels than Tougu Xiaotong Granule water extract serum group, Tougu Xiaotong Granule alcohol extract group (P < 0.01). Sox9 expression in Tougu Xiaotong Granule water extract serum and chondroinductive group were significantly higher than that in the chondroinductive group. Experimental findings indicate that, Tougu Xiaotong Granule containing serum can accelerate bone marrow stromal stem cells differentiate into cartilage cells by up-regulation of Sox9 expression.

Key words: stem cells, stem cells and traditional Chinese medicine, osteoarthritis, Tougu Xiaotong Granule, bone marrow mesenchymal stem cells, cartilage differentiation, drug-containing serum, water extract, alcohol extract, invigorate the kidney and promote blood flow, cartilage cells, seed cells, National Natural Science Foundation of China, stem cell photographs-containing paper

CLC Number:

R394.2

Liu Bo-ling, Li Xi-hai, Xiao Li-chun, Liang Gui-qing, Wu Guang-wen, Li Zhao-hui, Chen Qi-yong . Tougu Xiaotong Granule containing serum induces cartilage differentiation of bone marrow stromal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(32): 5863-5870.

Figures/Tables 9

References

[1] 李西海,刘献祥. 透骨消痛颗粒的现代药理学基础[J].福建中医药, 2006,37(1):50-52.

[2] 吴追乐,李西海,吴广文,等.透骨消痛胶囊含药血清对软骨细胞线粒体凋亡通路的影响[J].中华中医药杂志,2011,26(2): 343-346.

[3] 刘伯龄,张锡庆. hVEGF165基因转染后骨髓间充质干细胞蛋白分泌功能及成骨活性的检测[J].中国组织工程研究与临床康复, 2007,11(46):9242-9245.

[4] 刘伯龄,张锡庆.血管内皮细胞生长因子基因转染骨髓基质细胞的瞬时表达与稳定表达[J].中国微循环,2008,12(1):16-19.

[5] Pradier A, Ettelaie C. The influence of exogenous tissue factor on the regulators of proliferation and apoptosis in endothelial cells. J Vasc Res. 2008;45(1):19-32.

[6] Erggelet C, Neumann K, Endres M, et al. Regeneration of ovine articular cartilage defects by cell-free polymer-based implants.Biomaterials.2007;28(36):5570-5580.

[7] 刘伯龄,邹吉林,梁珪清,等. 透骨消痛颗粒对关节软骨细胞wnt/β-连环蛋白信号通路的调控[J].中国组织工程研究与临床康复,2011,15(46):8574-8578.

[8] 刘伯龄,梁硅清,刘献祥,等.透骨消痛颗粒干预膝骨性关节炎早期环氧合酶2和诱导型一氧化氮合酶的表达[J].中国组织工程研究与临床康复,2011,15(11):2034-2037.

[9] 刘伯龄,高雪伟,梁圭清,等.早期膝骨性关节炎NO炎症途径及中药干预的实验研究[J].中国中医骨伤科杂志,2010,18(8):3- 5.

[10] Collier ME, Ettelaie C. Induction of endothelial cell proliferation by recombinant and microparticle-tissue factor involves beta1-integrin and extracellular signal regulated kinase activation. Arterioscler Thromb Vasc Biol. 2010;30(9): 1810-1807.

[11] Krüger JP, Hondke S, Endres M, et al. Human platelet-rich plasma stimulates migration and chondrogenic differentiation of human subchondral progenitor cells. J Orthop Res. 2012; 30(6):845-852.

[12] Aghajanova L, Horcajadas JA, Esteban FJ, et al. The bone marrow-derived human mesenchymal stem cell: potential progenitor of the endometrial stromal fibroblast. Biol Reprod. 2010;82(6):1076-1087.

[13] Richardson SM, Hoyland JA, Mobasheri R, et al. Mesenchymal stem cells in regenerative medicine: opportunities and challenges for articular cartilage and intervertebral disc tissue engineering. J Cell Physiol. 2010;222(1):23-32.

[14] Huselstein C, Li Y, He X. Mesenchymal stem cells for cartilage engineering. Biomed Mater Eng. 2012;22(1-3): 69-80.

[15] 叶蕻芝,李西海,梁文娜,等.透骨消痛颗粒含药血清对兔软骨细胞增殖影响的研究[J].中国临床药理学与治疗学, 2009,14(6): 624-628.

[16] 梁祖建,韩清民,张还添.“补肾活血方”对骨关节炎软骨保护的效应与机制研究[J].江苏中医药,2009,4(3):33-34.

[17] 林木南,刘献祥. 透骨消痛方治疗骨性关节炎30例[J].福建中医药,2005,36(4):15-16.

[18] 李民,陈文列,刘献祥,等.透骨消痛胶囊干预膝骨性关节炎软骨下骨重塑分子机制探讨[J].福建中医学院学报,2009,19(6): 35-39.

[19] 刘献祥,李西海,周江涛,等.透骨消痛颗粒防治膝骨性关节炎的机理研究[J].中国中西医结合杂志,2007,27(1):50-54.

[20] Amano K, Hata K, Sugita A,et al. Sox9 family members negatively regulate maturation and calcification of chondrocytes through up-regulation of parathyroid hormone-related protein. Mol Biol Cell. 2009;20(21): 4541-4551.

[21] Csaki C, Schneider PR, Shakibaei M. Mesenchymal stem cells as a potential pool for cartilage tissue engineering. Ann Anat. 2008;190(5):395-412.

[22] Li H, Haudenschild DR, Posey KL, et al. Comparative analysis with collagen type II distinguishes cartilage oligomeric matrix protein as a primary TGFβ-responsive gene.Osteoarthritis Cartilage.2011;19(10):1246-1253.

[23] Yang HN, Park JS, Woo DG, et al.Chondrogenesis of mesenchymal stem cells and dedifferentiated chondrocytes by transfection with SOX Trio genes. Biomaterials. 2011;32 (30):7695-7704.