低水平激光辐射通过 APN/Wnt/β-连环蛋白通路促进骨髓基质细胞向成骨细胞分化。
Low-level laser irradiation promotes the differentiation of bone marrow stromal cells into osteoblasts through the APN/Wnt/β-catenin pathway.
机构信息
Department of Orthopaedics, the 89th Hospital of People's Liberation Army, Weifang, China.
出版信息
Eur Rev Med Pharmacol Sci. 2018 May;22(9):2860-2868. doi: 10.26355/eurrev_201805_14988.
OBJECTIVE
The relationship between adiponectin (APN) pathway and Wnt pathway was explored through BMSCs, and the effect of low-level laser irradiation (LLLI) on bone marrow stromal cells (BMSCs) and its mechanism were further studied.
MATERIALS AND METHODS
3-week-old Sprague-Dawley (SD) rats were selected, and mesenchymal stem cells were separately cultured and purified. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to analyze cell proliferation. After osteogenic and adipogenic induction, cultures were conducted, respectively, cells were stained with alizarin red and oil red O. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expressions of osteogenesis-related genes, runt-related transcription factor 2 (RUNX2), and osteocalcin (OC) and those of adipogenesis-related genes, peroxisome proliferator-activated receptor-gamma (PPARγ) and CCAAT/enhancer-binding protein alpha (c/EBPα). Western blotting was used to detect the expressions of β-catenin in the cytoplasm and nucleus. The lentiviral expression vector of adiponectin receptors (APN-R) was constructed, and the expression of APN receptor genes was silenced. The expressions of β-catenin in APN receptors and the nucleus within cells were detected.
RESULTS
LLLI promoted the bone formation by inducing the differentiation direction of mesenchymal stem cells, increasing the number of osteoblasts in the bone marrow and inhibiting the reduction of the number of adipocytes. LLLI regulates the Wnt pathway, promotes the entry of β-catenin into the nucleus, activates the osteogenic effect of the Wnt pathway so as to promote the bone formation of osteoblasts and inhibit bone resorption of osteoclasts. LLLI promotes the entry of β-catenin into the nucleus and the osteogenic differentiation of BMSCs through the APN pathway.
CONCLUSIONS
In summary, LLLI can promote osteogenesis and inhibit adipocytes formation, thus attenuating bone resorption of osteoclasts. The mechanism of LLLI is that it promotes the entry of β-catenin into the nucleus and regulates the Wnt pathway and the differentiation direction of mesenchymal stem cells through the APN signal pathway, thus promoting bone formation.
目的
通过骨髓间充质干细胞(BMSCs)探索脂联素(APN)通路与 Wnt 通路的关系,并进一步研究低水平激光照射(LLLI)对骨髓基质细胞(BMSCs)的影响及其机制。
材料和方法
选择 3 周龄的 Sprague-Dawley(SD)大鼠,分别培养和纯化间充质干细胞。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法分析细胞增殖情况。在成骨和脂肪诱导后,分别进行细胞染色,用茜素红和油红 O 染色。采用逆转录-聚合酶链反应(RT-PCR)检测成骨相关基因 runt 相关转录因子 2(RUNX2)和骨钙素(OC)以及脂肪形成相关基因过氧化物酶体增殖物激活受体-γ(PPARγ)和 CCAAT/增强子结合蛋白α(c/EBPα)的表达。采用 Western blot 检测细胞质和细胞核中β-连环蛋白的表达。构建脂联素受体(APN-R)的慢病毒表达载体,沉默 APN 受体基因,检测细胞内 APN 受体和细胞核中β-连环蛋白的表达。
结果
LLLI 通过诱导间充质干细胞的分化方向,增加骨髓中成骨细胞的数量,抑制脂肪细胞数量的减少,从而促进骨形成。LLLI 调节 Wnt 通路,促进β-连环蛋白进入细胞核,激活 Wnt 通路的成骨作用,从而促进成骨细胞的骨形成和抑制破骨细胞的骨吸收。LLLI 通过 APN 通路促进β-连环蛋白进入细胞核和 BMSCs 的成骨分化。
结论
综上所述,LLLI 可促进成骨和抑制脂肪细胞形成,从而减轻破骨细胞的骨吸收。LLLI 的作用机制是通过 APN 信号通路促进β-连环蛋白进入细胞核,调节 Wnt 通路和间充质干细胞的分化方向,从而促进骨形成。