红景天苷对诱导的成骨细胞在低氧环境中的保护作用

摘要
图/表
参考文献
相关文章 (15)

全文: PDF (1420 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要目的：探讨红景天苷对诱导的成骨细胞在低氧环境中的保护作用及其机制。方法：组织块法分离培养小鼠成纤维细胞，转染重编程为多能诱导干细胞，进行Nanog、Oct-4、Rexl、Sox-2的QT-PCR检测。诱导性多潜能干细胞（IPS细胞）诱导分化为成骨细胞，进行茜素红染色鉴定。将诱导后的成骨细胞分为红景天苷预处理组、正常组和低氧组，用Annexin V-FITC/PI染色检测细胞凋亡率，Western blot检测HIF-1α和VEGF蛋白表达量。结果：多能诱导干细胞的内源性基因Nanog、Oct-4、Rexl、Sox-2的表达量均明显高于小鼠成纤维细胞（均P＜0.05）。茜素红染色显示诱导后的成骨细胞染色阳性。红景天苷预处理组细胞凋亡率明显低于低氧组，差异有统计学意义（P＜0.05）。红景天苷预处理组的HIF-1α和VEGF表达量高于低氧组（0.93±0.05 vs. 0.81±0.02，0.95±0.03 vs. 0.79±0.04；均P＜0.05）。结论：红景天苷对诱导后成骨细胞在低氧环境中有保护作用，其机制可能是通过提高细胞HIF-1α和VEGF的表达来抑制成骨细胞的凋亡。

	服务
	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	沈新升
	陈鸥
	赖宪良
	苏嘉

关键词 ：红景天苷, 多能诱导干细胞, 低氧, 成骨细胞, 凋亡

Abstract：Objective: To investigate the protective effect of salidroside on induced osteoblasts in hypoxia environment and its mechanism. Methods: the mouse fibroblasts were separated and cultured by tissue block method. The transfected induced pluripotent stem cells were reprogrammed and the QT-PCR of Nanog, Oct-4, Rexl and Sox-2 were detected to the induced osteoblasts differentiated from the induced pluripotent stem cells and identified by alizarin red staining. The induced osteoblasts were divided into three groups: salidroside preconditioning group, normal oxygen group and hypoxia group. Annexin V-FITC/PI staining was used to detect the apoptosis rate, and Western-blot was used to detect VEGF, HIF-1 alpha. Results: The expression of endogenous gene Nanog, Oct-4, Rexl and Sox-2 of induced pluripotent stem cells was higher than that of mouse fibroblasts cells. The result is Nanog (783±42 vs. 308±23), Oct-4 (774±37 vs. 319±31), Rexl (742±28 vs. 382±31), Sox-2 (831±31 vs. 394±12). Alizarin red staining showed that the induced osteoblasts cells had positive staining results. The apoptotic rate of salidroside group was significantly lower than that of hypoxia group [(35.65±3.92)% vs. (95.20±1.03)%]. There was significant difference between the two groups (P<0.05). The expression of HIF-1a and VEGF in salidroside group was higher than that in hypoxic group (0.93±0.05 vs. 0.81±0.02, 0.95±0.03 vs. 0.79±0.04, P<0.05). Conclusion: Salidroside has protective effect on hypoxic environment of osteoblasts.

Key words： salidroside induced pluripotent stem cells hypoxia osteoblast cells apoptosis

收稿日期: 2018-06-11

基金资助:温州市公益性科技计划项目（Y20140594）。

作者简介: 沈新升（1963-），男，浙江金华人，副主任医师。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2019/V49/I3/179

[1] THUAKSUBAN N, MONMATURAPOJ N, LUNTHENG T. Effects of polycaprolactone-biphasic calcium phosphate scaffolds on enhancing growth and differentiation of osteo-blasts[J]. Biomed Mater Eng, 2018, 29(2): 159-176.
[2] POSRITONG S, HONG J M, ELENISTE P P, et al. Pyk2 deficiency potentiates osteoblast differentiation and mineralizing activity in response to estrogen or raloxifene[J]. Mol Cell Endrcrinol, 2018, 474: 35-47.
[3] ALTMAYER N C, GALATA V, WARSCHBURGER N, et al.
Gene amplification in mesenchymal stem cells and during differentiation towards adipocytes or osteoblasts[J]. Oncotarget, 2017, 9(2): 1803-1812.
[4] SHEYN D, BEN-DAVID S, SHAPIRO G, et al. Human induced pluripotent stem cells differentiate into functional mesenchymal stem cells and repair bone defects[J]. Stem Cells Transl Med, 2016, 5(11): 1447-1460.
[5] KATO H, OCHIAI-SHINO H, ONODERA S, et al. Promoting effect of 1,25(OH)2 vitamin D3 in osteogenic differentiation from induced pluripotent stem cells to osteocyte-like cells[J]. Open Biol, 2015, 5(2): 140201.
[6] MA M S, KANNAN V, DE VRIES A E, et al. Characterization and comparison of osteoblasts derived from mouse embryonic stem cells and induced pluripotent stem cells[J]. J Bone Miner Metab, 2017, 35(1): 21-30.
[7] TOKITA R, NAKAJIMA K, INOUE K, et al. Differentiation behavior of iPS cells cultured on PLGA with osteoinduction medium[J]. Dent Mater J, 2017, 36(1): 103-110.
[8] 祁琳, 王川, 梅国华, 等. 低氧状态下红景天苷经HIF-1α信号通路调节人成骨样MG-63细胞表型表达的研究[J]. 中国药理学通报, 2017, 33(6): 836-843.
[9] OKAMOTO H, MATSUMI Y, HOSHIKAWA Y, et al. Involvement of microRNAs in regulation ofosteoblastic differentiation in mouse induced pluripotent stemcells[J]. PLoS One, 2012, 7(8): e43800.
[10] 钱浩, 徐增琦, 李杨, 等, 小鼠诱导性多能干细胞诱导成骨分化的初步研究[J]. 口腔生物医学, 2015, 6(4): 193-197.
[11] GENG Y J. Molecular mechanisms for cardiovascular stem cell apoptosis and growth in the hearts with atherosclerotic coronary disease and ischemic heart failure[J]. Ann N Y Acad Sci, 2003, 687-697.
[12] FOLLMAR K E, DECROOS F C, PRICHARD H L, et al. Effects of glutamine, glucose, and oxygen concentration on the metabolism and proliferation of rabbit adipose-derived stem cells[J]. Tissue Eng, 2006, 12(12): 3525-3533.
[13] TAN C B, GAO M, XU W R, et al. Protective effects of salidroside on endothelial cell apoptosis induced by cobalt chlo-ride[J]. Biol Pharm Bull, 2009, 32(8): 1359-1363.
[14] 李庆勇, 钱志远, 卞中国, 等. 创伤性颅脑损伤后缺氧诱导因子-1α的表达及其在血管修复中的作用[J]. 中华实验外科杂志, 2015, 32(4): 926.
[15] STEGEN S, VAN GASTEL N, EELEN G, et al. HIF-1α promotes glutamine-mediated redox homeostasis and glycogen-dependent bioenergetics to support postimplantation bone cell[J]. Cell Metab, 2016, 23(2): 265-279.
[16] LIN Y, ZHAI E, LIAO B, et al. Autocrine VEGF signaling promotes cell proliferation through a PLC-dependent pathway and modulates Apatinib treatment efficacy in gastric cancer[J]. Oncotarget, 2017, 8(7): 11990-12002.
[17] 郦铮铮, 林以诺, 郭宴会, 等, VEGF经PI3K-Akt-Bad通路降低冻存后EOCs凋亡率[J]. 温州医科大学学报, 2014, 44(12): 882-886.
[18] KLUCK R M, BOSSY-WETZEL E, GREEN D R, et al. The release of cytochrome c from mitochondria: A primary site for bcl-2 regulation of apoptosis[J]. Science, 1997, 275(5303): 1132-1136.