氯化锂促进骨髓间充质干细胞增殖分化的作用途径  

Lithium chloride promotes proliferation and differentiation of bone marrow mesenchymal stem cells via Wnt/beta-catenin signaling pathway

作　　者：付殷[1] 孙贵才[2] 涂远青[2] 凌小鹏[2] Fu Yin;Sun Guicai;Tu Yuanqing;Ling Xiaopeng(Heilongjiang University of Traditional Chinese Medicine,Harbin 150036, Heilongjiang Province, China;Department of Orthopedics, Fourth Affiliated Hospital of NanchangUniversity, Nanchang 330003, Jiangxi Province, China)

机构地区：[1]黑龙江中医药大学,黑龙江省哈尔滨市150036 [2]南昌大学第四附属医院骨科,江西省南昌市330003

出　　处：《中国组织工程研究》2019年第25期3956-3960,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81473503),项目负责人:孙贵才~~

摘　　要：背景:寻找促进骨髓间充质干细胞增殖及向成骨细胞分化的方法,可以为治疗骨质疏松类疾病提供治疗思路。目的:探讨氯化锂调控大鼠骨髓间充质干细胞增殖分化的机制。方法:实验采用全骨髓细胞接种法和贴壁纯化培养大鼠骨髓间充质干细胞,分别给予0,2,5 nmol/L氯化锂进行干预;采用CCK-8法检测大鼠骨髓间充质干细胞的增殖情况,茜素红染色法检测钙化结节形成情况,PNPP法测定碱性磷酸酶活性,Western blot法检测大鼠骨髓间充质干细胞中GSK3β,p-GSK3β,β-catenin蛋白含量变化。结果与结论:①2,5 nmol/L氯化锂均能促进大鼠骨髓间充质干细胞增殖,5 nmol/L氯化锂促增殖能力最强;②2,5 nmol/L氯化锂组矿化结节与碱性磷酸酶活性均比0 nmol/L氯化锂组明显,且5 nmol/L氯化锂促成骨分化更为显著;③2,5 nmol/L氯化锂能够显著增加p-GSK3β、β-catenin蛋白表达,而各组GSK3β蛋白表达差异无显著性意义;④结果表明,氯化锂能够促进大鼠骨髓间充质干细胞的增殖及向成骨细胞分化,其可能机制是通过调控Wnt/β-catenin信号通路相关蛋白表达实现的。BACKGROUND: Seeking proper methods to promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells can provide therapeutic ideas for the treatment of osteoporosis.OBJECTIVE: To study the mechanism of lithium chloride on the proliferation and differentiation of rat bone marrow mesenchymal stem cells.METHODS: Rat bone marrow mesenchymal stem cells were cultured with the whole bone marrow cell inoculation and adherent purification, followed by intervention with 0, 2, 5 nmol/L lithium chloride. Cell counting kit-8 was used to detect the effect of lithium chloride on the proliferation of bone marrow mesenchymal stem cells. Alizarin red staining was used to determine calcified nodules.PNPP method was applied to measure alkaline phosphatase activity. The contents of glycogen synthase kinase 3β, phosphorylated glycogen synthase kinase 3β and β-catenin in rat bone marrow mesenchymal stem cells were determined by western blot.RESULTS AND CONCLUSION: Lithium chloride, 2 and 5 nmol/L, promoted the proliferation of bone marrow mesenchymal stem cells and5 nmol/L lithium chloride had the strongest proliferative effect. Compared with 0 nmol/L group, more calcified nodules and higher alkaline phosphatase activity were observed in the 2 and 5 nmol/L groups, especially in the 5 nmol/L group. Both 2 and 5 nmol/L lithium chloride upregulated the expression of phosphorylated glycogen synthase kinase 3β and β-catenin proteins, but showed no difference in the expression of glycogen synthase kinase 3β. In conclusion, lithium chloride can promote the proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells, and the possible mechanism is through the regulation of Wnt/β-catenin signaling pathway-related protein expression.

关 键 词：氯化锂 骨髓间充质干细胞 细胞增殖 成骨分化 钙化结节 碱性磷酸酶活性 WNT/Β-CATENIN信号通路 国家自然科学基金 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]