Effect of Compound Jiegu Tablets and Zhuanggu Tablets on thedifferentiation of bone marrow mesenchymal stem cells in vitro

doi:10.3969/j.issn.2095-4344.2017.33.003

Abstract

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are a kind of pluripotent stem cells in the bone marrow. Drug-containing serum treatment may be followed by a variation in the expression of genes involved in multiple signaling pathways.
OBJECTIVE: To study the effect of compound Jiegu Tablets and Zhuanggu Tablets on the differentiation of BMSCs in vitro.
METHODS: Sixty Sprague-Dawley rats were randomized into Jiegu group, Zhuanggu group, combined group and blank control group. Different drug-containing sera were prepared and used to culture BMSCs isolated from the rat bone marrow. RT-PCR and western blot assay were used to detect the expression of BMP-2, Runx2, VEGF and Akt at mRNA and protein levels, respectively, at 24 hours after culture.
RESULTS AND CONCLUSION: Compared with the blank control group, the expression levels of Akt and VEGF mRNA and protein were significantly increased in the Jiegu group and combined group (P < 0.05 or P < 0.01), but showed no difference in the Zhuanggu group (P > 0.05). Compared with the blank control group, the expression levels of BMP-2 and Runx2 mRNA and protein were significantly increased in the Jiegu group (P < 0.05) and very significantly increased in the Zhuanggu group and combined group (P < 0.01). These findings indicate that compound Jiegu Tablets and Zhuanggu Tablets containing sera lead to synergistic effects on theBMSCs differentiation into vascular endothelial cells and osteoblasts in vitro.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Bone Marrow, Mesenchymal Stem Cells, Cell Differentiation, Serum, Tissue Engineering

CLC Number:

R394.2

Wu Bo, Liu Ye, Ma Xu, Liu Su-yuan, Wang Yi-han, Wang Jia-yuan, Tan Cheng-bo, Yang Zheng-bo. Effect of Compound Jiegu Tablets and Zhuanggu Tablets on thedifferentiation of bone marrow mesenchymal stem cells in vitro[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(33): 5262-5267.

Figures/Tables 2

References

[1] Zeng YL, Zheng H, Chen QR, et al. Bone marrow-derived mesenchymal stem cells overexpressing MiR-21 efficiently repair myocardial damage in rats. Oncotarget. 2017;8(17): 29161-29173.

[2] Liu L, Luo Q, Sun J, et al. Decreased nuclear stiffness via FAK-ERK1/2 signaling is necessary for osteopontin-promoted migration of bone marrow-derived mesenchymal stem cells. Exp Cell Res. 2017;355(2):172-181.

[3] Long R, Liu Y, Wang S, et al. Co-microencapsulation of BMSCs and mouse pancreatic β cells for improving the efficacy of type I diabetes therapy. Int J Artif Organs. 2017;40(4):169-175.

[4] Wu JH, Thoreson AR, Gingery A, et al. The revitalisation of flexor tendon allografts with bone marrow stromal cells and mechanical stimulation: An ex vivo model revitalising flexor tendon allografts. Bone Joint Res. 2017;6(3):179-185.

[5] Li D, Zhu L, Liu Y, et al. Stable subcutaneous cartilage regeneration of bone marrow stromal cells directed by chondrocyte sheet. Acta Biomater. 2017;54:321-332.

[6] Zhu YL, Wang S, Ding DG, et al. miR?217 inhibits osteogenic differentiation of rat bone marrow?derived mesenchymal stem cells by binding to Runx2. Mol Med Rep. 2017;15(5): 3271-3277.

[7] Bottagisio M, Lopa S, Granata V, et al. Different combinations of growth factors for the tenogenic differentiation of bone marrow mesenchymal stem cells in monolayer culture and in fibrin-based three-dimensional constructs. Differentiation. 2017;95:44-53.

[8] Cleary MA, Narcisi R, Albiero A, et al. Dynamic Regulation of TWIST1 Expression During Chondrogenic Differentiation of Human Bone Marrow-Derived Mesenchymal Stem Cells. Stem Cells Dev. 2017;26(10):751-761.

[9] 赵保胜,李兰芳,马悦颖,等.参苏饮含药血清对小鼠巨噬细胞Toll样受体及其下游信号转导通路的影响[J].中国中药杂志, 2007, 32(4): 327-332.

[10] 苏友新,闫虎,陈宝军,等.壮骨健膝方含药血清中药物成分骨碎补柚皮苷对IL-1β诱导兔退变软骨细胞“caveolin-p38MAPK”信号通路的影响[J].中国中西医结合杂志,2014,34(12):1492-1498.

[11] 柳新,薛玲,高杰,等.白香丹含药血清对大鼠海马神经元中5-羟色胺2C受体的表达及其下游信号通路中IP3的影响[J].中国药理学通报, 2012, 28(7): 1019-1023.

[12] 赵保胜,刘洪斌,马悦颖,等.桂枝汤含药血清对小鼠巨噬细胞Toll样受体3、4型及其下游信号转导通路元件的影响[J].中药药理与临床, 2007, 23(3):1-4.

[13] Niu YB, Kong XH, Li YH, et al. Radix Dipsaci total saponins stimulate MC3T3-E1 cell differentiation via the bone morphogenetic protein-2/MAPK/Smad-dependent Runx2 pathway. Mol Med Rep. 2015;11(6):4468-4472.

[14] Hu B, Yu B, Tang D, et al. Daidzein promotes osteoblast proliferation and differentiation in OCT1 cells through stimulating the activation of BMP-2/Smads pathway. Genet Mol Res. 2016;15(2):gmr8792.

[15] Chen B, Zhang F, Li QY, et al. Protective Effect of Ad-VEGF-Bone Mesenchymal Stem Cells on Cerebral Infarction.Turk Neurosurg. 2016;26(1):8-15.

[16] Lin SS, Zhu B, Guo ZK, et al. Bone marrow mesenchymal stem cell-derived microvesicles protect rat pheochromocytoma PC12 cells from glutamate-induced injury via a PI3K/Akt dependent pathway. Neurochem Res. 2014;39(5):922-931.

[17] Bateman ME, Strong AL, Hunter RS, et al. Osteoinductive effects of glyceollins on adult mesenchymal stromal/stem cells from adipose tissue and bone marrow. Phytomedicine. 2017;27:39-51.

[18] 徐练,孔清泉.调控成骨细胞分化及骨形成关键信号通路的研究进展[J].中国修复重建外科杂志, 2014,28(12):1484-1489.

[19] 王国红,李祺福. Cbfa1/Runx2与成骨细胞分化调控[J].生命科学, 2005, 17(1): 40-44.

[20] 王慧,李玉坤.成骨细胞分化调控因子研究进展[J].国际骨科学杂志, 2011, 32(6): 377-379.

[21] La WG, Jin M, Park S, et al. Delivery of bone morphogenetic protein-2 and substance P using graphene oxide for bone regeneration. Int J Nanomedicine. 2014;9 Suppl 1:107-116.

[22] 刘光源,田发明,张柳,等. BMP2信号通路与经典Wnt信号通路及相互交联对成骨分化的调控[J].中国骨质疏松杂志, 2014,20(5): 551-556.

[23] Chen Y, Whetstone HC, Youn A, et al. Beta-catenin signaling pathway is crucial for bone morphogenetic protein 2 to induce new bone formation. J Biol Chem. 2007;282(1):526-533.

[24] Phimphilai M, Zhao Z, Boules H, et al. BMP signaling is required for RUNX2-dependent induction of the osteoblast phenotype. J Bone Miner Res. 2006;21(4):637-646.

[25] Leboy P, Grasso-Knight G, D'Angelo M, et al. Smad-Runx interactions during chondrocyte maturation. J Bone Joint Surg Am. 2001;83-A Suppl 1(Pt 1):S15-22..

[26] 毕锴,温建民,孙永生,等.中医药对骨折愈合作用及其机制的实验研究进展[J].中国中医基础医学杂志, 2014,20(1): 137-140.

[27] 尹朝献,刘安平,曹日隆.中药促进骨折愈合机制的实验研究进展[J].中医药临床杂志, 2007, 19(6): 640-642.

[28] 卫鹏斌,张民,李军,等. VEGF在BMP-2 促进骨质疏松性成骨细胞过程中作用机制的研究[J].中国临床实用医学, 2015, 6(3): 13-16.

[29] Zhou L, Lin Q, Wang P, et al. Enhanced neuroprotective efficacy of bone marrow mesenchymal stem cells co-overexpressing BDNF and VEGF in a rat model of cardiac arrest-induced global cerebral ischemia. Cell Death Dis. 2017;8(5):e2774.

[30] Kang ML, Kim JE, Im GI.Vascular endothelial growth factor-transfected adipose-derived stromal cells enhance bone regeneration and neovascularization from bone marrow stromal cells. J Tissue Eng Regen Med. 2017 Feb 15. [Epub ahead of print]

[31] 安福,王永刚,刘文忠,等. VEGF在大鼠脊髓损伤后股骨干骨痂形成过程中的表达[J].医学综述, 2014, 20(1): 129-131,135.

[32] Nishi J, Minamino T, Miyauchi H, et al. Vascular endothelial growth factor receptor-1 regulates postnatal angiogenesis through inhibition of the excessive activation of Akt. Circ Res. 2008;103(3):261-268.

[33] Zhang Q, Zhao Y, Xu Y, et al. Sodium ferulate and n-butylidenephthalate combined with bone marrow stromal cells (BMSCs) improve the therapeutic effects of angiogenesis and neurogenesis after rat focal cerebral ischemia. J Transl Med. 2016;14(1):223.

[34] 邬波,马旭,柳椰,等.复方接骨片含药血清对BMP-2沉默的骨髓间充质干细胞体外增殖及分化影响[J].辽宁中医药大学学报, 2015, 17(9): 5-8.

[35] 刘建群,王雪梅,刘一文,等.基于影响药物代谢的中药配伍研究进展[J].中国实验方剂学杂志, 2014, 20(19): 221-224.

[36] 董芊芊,王四旺.分子中药学和分子配伍理论[J].临床医药文献电子杂志, 2014, 2(6): 124-124.

[37] 邬波,马旭,柳椰,等.不同中药制剂治疗骨折临床观察[J].辽宁中医药大学学报, 2014, 16(6): 5-7.

[38] 邬波,柳椰,马旭,等.不同中药制剂治疗骨质疏松的临床观察[J]. 中华中医药学刊, 2016, 34(11): 2567-2569.