恒古骨伤愈合剂通过TGFβ/Smad信号通路促进去势骨质疏松症大鼠BMSC增殖

郑红; 李进涛; 袁鑫; 吴超; 曾河; 赵宏斌; 唐薇

恒古骨伤愈合剂通过TGFβ/Smad信号通路促进去势骨质疏松症大鼠BMSC增殖

郑红¹,
李进涛¹,
袁鑫²,
吴超¹,
曾河¹,
赵宏斌²,
唐薇²

1. 昆明医科大学实验动物学部
2. 昆明医科大学第一附属医院急诊医学部

基金项目:

基金：国家自然科学基金资助项目 (81460647); 云南省应用基础研究计划项目 (2014FB033;2017FE-014&-016);

计量
- 文章访问数: 2412
- HTML全文浏览量: 883
- PDF下载量: 167
- 被引次数: 0
出版历程
- 收稿日期: 2017-10-21

Osteoking Promotes the Proliferation of BMSC through Activating TGF-β/Smad Signaling Pathway in Castrate Osteoporosis Rats

Funds:

基金：国家自然科学基金资助项目 (81460647); 云南省应用基础研究计划项目 (2014FB033;2017FE-014&-016);

摘要

摘要: 目的研究恒古骨伤愈合剂 (OK) 通过TGFβ/Smad信号通路对骨质疏松 (OP) 大鼠骨髓基质干细胞 (BMSCs) 细胞分化的作用.方法摘除大鼠双侧卵巢, 复制OP模型, 分离、培养BMSCs.将源于假手术大鼠的BMSCs常规培养作为正常对照组, 将源于OP大鼠的BMSCs分为5组, OP模型组常规培养, 阳性对照组加入1μmol/L阿仑膦酸钠, OK低、中、高治疗组分别加入恒古骨伤愈合剂50 mL/L、100 mL/L和200 mL/L, 孵育24 h, 收集各组细胞.采用MTT法检测BMSCs增殖率, ELISA法检测细胞骨形成标志物含量, RT-qPCR法检测TGF-β的m RNA表达水平, Western blot法检测TGF-β、p-Smad2/3以及Smad2/3的蛋白表达水平.结果与正常对照组比较, OP模型对照组BMSCs细胞增殖率降低, 骨形成标志物OC、PINP和BALP含量降低, TGF-β的m RNA和蛋白表达水平降低, Smad2/3的磷酸化水平降低, 差异有统计学意义 (P<0.05) .经OK治疗后, 与模型组比较, 上述均有不同程度缓解, OK组疗效呈现出剂量依赖性, 且1μmol/L阿仑膦酸钠、100mL/L和200 mL/L OK组治疗效果均有统计学意义 (P<0.05) .结论 OK可以通过激活TGF-β/Smad信号通路促进成骨细胞增殖, 从而达到治疗绝经后OP的效果.
- 恒古骨伤愈合剂 /
- 骨质疏松 /
- 骨髓基质干细胞 /
- TGF-β/Smad信号通路
Abstract: Objective To study the effect of OK on the differentiation of bone marrow stromal stem cells (BMSCs) through the TGFβ/Smad signaling pathway in rats. Methods We removed the bilateral ovaries of rats to replicate OP model, and isolated and cultured BMSCs. BMSCs isolated from nomal rats were cultured as control group, BMSCs isolated from OP rats were divided into 5 groups, OP model group was regularly cultured, positive control group was treated with alendronate sodium (1 μmol/L) , low, medium and high OK treatment group were treated with 50 mL/L, 100 mL/L and 200 ml/L OK respectively.After 24 h incubation, all cells were collected. The proliferation rate of BMSCs was determined by MTT method, and ELISA method was employed to detect the contents of bone formation markers, RT-qPCR was used to determine the m RNA expression level of TGFβ and the protein expression levels of TGFβ, p-Smad2/3 and of Smad2/3 were detected by Western blot. Results Compared with control group, the proliferation rate of the BMSCs in OP model group was reduced, concentrations of bone formation markers (OC, PINP and BALP) were reduced, m RNA and protein expression levels of TGFβ, as well as the phosphorylated level of Smad2/3 were downregulated.The difference was statistically significant (P<0.05) . After treatment with OK, compared with model group, all the above effects were ameliorated in different degree, a dose dependent manner was observed in OK treatment group, and the treatment effects of alendronate sodium (1 μmol/L) , 100 mL/L and 200 mL/L OK group were statistically significant (P <0.05) .Conclusion OK can promote the proliferation of osteoblasts by activating TGFβ/Smad signaling pathway to achieve the effect of treating postmenopausal OP.
- Osteoking /
- Osteoporosis /
- Bone marrow stromal stem cells /
- TGFβ/Smad signaling pathway

HTML全文

参考文献(15)

[1]姚荣成, 谢宇, 张悦, 等.JGYS对大鼠去卵巢高转换型骨质疏松症的影响[J].昆明医科大学学报, 2015, 36 (1) :4-11.
[2]	[2]MACLEAN C, NEWBERRY S, MAGLIONE M, et al.Systematic review:comparative effectiveness of treatments toprevent fractures in men and women with low bone densityor osteoporosis[J].Ann Intern Med, 2008, 148 (3) :197-213.
[3]	[3]VARELA A, CHOUINARD L, LESAGE E, et al.One year of abaloparatide, a selective peptide activator of the PTH1receptor, increased bonemass and strength in ovariectomized rats[J].Bone, 2016, 95 (2) :143-150.
[4]	[4]ADLER R A, EL-HAJJ F G, BAUER D C, et al.Managing osteoporosisin patients on long-term bisphosphonate treatment:report of a task force of the american society for bone and mineral research[J].Journal of Bone&Mineral Research the Official Journal of the American Society for Bone&Mineral Research, 2015, 31 (1) :16-35.
[5]于洋, 邵秉一, 周志斐, 等.骨髓间充质干细胞对雌激素缺乏所致骨质疏松症的治疗作用[J].细胞与分子免疫学杂志, 2013, 29 (12) :1267-1271.
[6]李丽, 张悦, 杨仁华, 等.老鹳草素对骨质疏松大鼠BMSCWnt3a表达的影响[J].昆明医科大学学报, 2016, 37 (6) :9-12.
[7]	[7]FAN J Z, YANG L, MENG G L, et al.Estrogen improvesthe proliferation and differentiation of h BMSCs derived frompostmenopausal osteoporosis through notch signaling pathway[J].Mol Cell Biochem, 2014, 392 (1-2) :85-93.
[8]袁鑫, 郑红, 角建林, 等.口服恒古骨伤愈药对骨质疏松性骨折树部分血液生化指标的影响[J].昆明医科大学学报, 2017, 38 (7) :12-15.
[9]王素彬, 赵宏斌, 胡敏, 等.恒古骨伤愈合剂对骨质疏松性骨折兔骨密度及DKK-1蛋白的影响[J].中国中西医结合杂志, 2015, 35 (8) :1000-1003.
[10]徐梦思, 黄涛, 刘丽娟, 等.TGFβ-SMAD信号通路基因在梅山猪与杜洛克猪不同级别卵泡中的表达分析[J].家畜生态学报, 2016, 37 (2) :12-18.
[11]	[11]HERLUND E, SVEDBOM A, IVERGARD M, et al.Osteoporosis in the European Union:medical management, epidemiology and economic burden[J].Archives of Osteoporosis, 2013, 8 (1) :111-115.
[12]	[12]FLORESR, SHI J, FUHRMAN B, et al.Fecal microbial determinants offecal and systemic estrogens and estrogen metabolites:a crosssectional study[J/OA].J Transl Med, 2012, (10) :253.
[13]秦桂美.兔BMSC_s体外培养及诱导为成骨细胞的研究[J].中医临床研究, 2011, 19 (3) :105-106.
[14]林凯, 虞德兵, 解晓东, 等.TGF-β/Smad信号通路在Follistatin调节鸭骨骼肌卫星细胞增殖过程中的作用机制[J].中国农业科学, 2015, 48 (12) :2460-2468.
[15]	[15]WILLIS B C, BROOK Z.TGF-βinduced EMT:mechanisms and implications for fibroticlungdisease[J].Am J Physiol Lung Cell Mol Physiol, 2007, 293 (12) :525-534.

相关文章(20)

[1]	冉蕾晶, 王绍华, 李林童, 桂莉. 益生菌辅助治疗骨质疏松患者效果的Meta分析, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20240209
[2]	邓绍友, 赵玉兰, 王佩锦, 李蓉, 李进涛, 郑红. 恒古骨伤愈合剂联合广谱抗生素改善db/db小鼠胰岛素抵抗和肠道菌群, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230530
[3]	邓绍友, 李蓉, 李进涛, 赵玉兰, 王佩锦, 郑红. 基于网络药理学探讨恒古骨伤愈合剂治疗骨关节炎的机制及动物实验初步验证, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230701
[4]	廖勇, 郭一雲, 周红宇, 冯海魄, 陈根定, 杨水清. 叶酸联合阿伦磷酸钠对绝经后骨质疏松患者骨代谢、CCL4及纤维细胞生长因子水平的影响, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20210716
[5]	季雨伟, 赵鑫, 陆姜利, 杨艺, 唐薇, 角建林. 恒古骨伤愈合剂对绝经后骨质疏松性骨折模型树鼩骨密度及骨生物力学的影响, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20201103
[6]	李娜, 方红丽, 邓苙. 原发性骨质疏松影响因素及健康干预研究进展, 昆明医科大学学报.
[7]	解春林, 岳桥宁, 滕兆伟. 骨质疏松疾病发生的一种非编码RNA科学假说, 昆明医科大学学报.
[8]	李丽, 李进涛, 郭玉倩, 吴超, 陆姜利, 曾跃勤, 角建林. 恒古骨伤愈合剂通过改善免疫功能调节骨质疏松骨折树鼩的骨重建, 昆明医科大学学报.
[9]	李丽, 曾跃勤, 李进涛, 赵宏斌, 梁张, 王利梅. 恒古骨伤愈合剂对骨质疏松树鼩肾组织CD₃₄和VEGF的影响, 昆明医科大学学报.
[10]	龙瑞清, 刘卓慧, 张帆, 李书聆, 余咏梅, 阮标. 绿脓假单胞菌感染中耳上皮细胞可以激活TGF-β1 Smad信号通路, 昆明医科大学学报.
[11]	邓苙, 李劲涛, 曹光琼. NGF-BMSCs移植对阿尔茨海默病大鼠神经行为学的改善, 昆明医科大学学报.
[12]	袁鑫, 郑红, 角建林, 吴超, 赵宏斌, 唐薇. 口服恒古骨伤愈药对骨质疏松性骨折树部分血液生化指标的影响, 昆明医科大学学报.
[13]	李进涛, 郑红, 李波, 袁鑫, 唐薇, 赵宏斌. 恒古骨伤愈合剂对骨质疏松骨折树鼩雌二醇和骨代谢的影响, 昆明医科大学学报.
[14]	赵正兴. 防旋型股骨近端髓内钉治疗骨质疏松性粗隆间骨折40例临床疗效, 昆明医科大学学报.
[15]	杨凤. 手术双侧卵巢去势法建立骨质疏松树鼩模型, 昆明医科大学学报.
[16]	雷雯. IL-27调控TGF-β/Smad通路在博来霉素诱导肺纤维化中的作用, 昆明医科大学学报.
[17]	李丽. 老鹳草素对骨质疏松大鼠BMSCWnt3a表达的影响, 昆明医科大学学报.
[18]	杨曼. 糖尿病SD大鼠骨密度及骨组织学特点研究, 昆明医科大学学报.
[19]	张小超. 老鹳草素对破骨细胞体外骨吸收功能的影响, 昆明医科大学学报.
[20]	宋宇锋. 去势大鼠关节软骨病理变化的研究, 昆明医科大学学报.