Mechanism of Osteoclast Differentiation Regulated by Akt-Gsk3β-Nfatc1 Signal Pathway Mediated by Gαi1/3 Protein
1.Department of Orthopaedicss，Affiliated Suqian Hospital of Xuzhou Medical University
2.Department of Orthopaedicss，Affiliated Hospital of Yangzhou University
Abstract：Objective To investigate the regulation of Gαi1/3 protein on osteoclast differentiation of bone marrow-derived macrophages (BMMs)and its mechanism. Methods Three BMMs，scr-shRNA，Gαi1/3-shRNA and Gαi1/3-DKO，were obtained by lentivirus knockdown and gene knockout strategies，and were treated with macrophage colony stimulating factor(MCSF)，receptor activator for nuclear factor-κB ligand (RANKL)and MCSF+RANKL for 30min.Western Blot was used to detect the expression of key proteins in the Akt-GSK3β-NFATc1 signaling pathway.MCSF and RANKL were combined to induce scr-shRNA，Gαi1/3shRNA BMMs 4 days.TRAP staining were used to observe the number of cell nuclei and the size and number of osteoclasts.Lentivirus scr-shRNA and Gαi1/3 shRNA were injected into the right epiphysis of the femoral shaft of a mouse model of osteoporosis to observe the reverse effect of Gαi1/3 knockdown on bone loss in mice.Results The activation of key proteins (p-Akt473，p-GSK3β，NFATc1)in the Akt-GSK3β-NFATc1 pathway induced by MCSF and MCSF+RANKL was inhibited by Gαi1/3.Knockdown of Gαi1/3 inhibits osteoclast differentiation and bone loss in mice in osteoporosis models.Conclusion Gαi1/3 protein regulates osteoclast differentiation by mediating Akt-GSK3β-NFATc1 pathway，which may provide a new idea and therapeutic target for the clinical treatment of osteoporosis.
［1］卢爱民，尹君，李春峰，等.运动疗法对绝经后骨质疏松症骨转换及炎性因子的影响［J］.实用骨科杂志，2023，29(5)：422-426.
［2］Zhang Y，Liu X，Li Y，et al.Aucubin slows the development of osteoporosis by inhibiting osteoclast differentiation via the nuclear factor erythroid 2 related factor 2 mediated antioxidation pathway［J］.Pharm Biol，2021，59(1)：1556-1565.
［3］Maxhimer JB，Bradley JP，Lee JC.Signaling pathways in osteogenesis and osteoclastogenesis：Lessons from cranial sutures and applications to regenerative medicine［J］.Genes Dis，2015，33(1)：57-68.
［4］褚赞波，邹荣鑫，黄海燕，等.破骨细胞功能调控与骨吸收抑制剂［J］.中华骨质疏松和骨矿盐疾病杂志，2018，11(5)：509-514.
［5］Marshall J，Zhou XZ，Chen G，et al.Antidepression action of BDNF requires and is mimicked by Gαi1/3 expression in the hippocampus［J］.Proc Natl Acad Sci U S A，2018，115(15)：E3549-E3558.
［6］Sun J，Huang W，Yang SF，et al.Galphai1 and Galphai3 mediate VEGF-induced VEGFR2 endocytosis，signaling and angiogenesis［J］.Theranostics，2018.8(17)：p.4695-4709.
［7］Shan HJ，Jiang K，Zhao MZ，et al.SCF/cKitactivated signaling and angiogenesis require Gαi1 and Gαi3［J］.Int J Biol Sci，2023，19(6)：1910-1924.
［8］柯呈辉，何立江，黄俊杰，等.利塞膦酸钠对骨质疏松性骨折大鼠愈合影响及生物力学评价［J］.实用骨科杂志，2019，25(6)：528-532.
［9］裴宝静，李建燕，辛立杰，等.福辛普利对去卵巢骨质疏松大鼠的治疗作用和机制研究［J］.实用骨科杂志，2020，26(11)：996-1001；1036.
［10］申浩，魏戌，谢雁鸣，等.绝经后骨质疏松症骨折危险因素及中医症状相关性研究［J］.中国中西医结合杂志，2017，37(1)：50-56.
［11］Crudden C，Shibano T，Song D，et al.Inhibition of G protein-coupled receptor kinase 2 promotes unbiased downregulation of IGF-1 receptor and restrains malignant cell growth［J］.Cancer Research，2020，81(2)：1662-2020.
［12］Wu M，Chen W，Lu Y，et al.Gα13 negatively controls osteoclast-ogenesis through inhibition of the Akt-GSK3β-NFATc1 signalling pathway［J］.Nat Commun，2017(8)：13700.
［13］李崇，罗晓婷，纪舒妤，等.炎症因子在骨质疏松发病机理中的研究进展［J］.中国骨质疏松杂志，2021，27(10)：1516-1522.
［14］李辉，谢兴文，李宁，等.细胞因子在骨质疏松症中的作用及中医药干预研究进展［J］.中国实验方剂学杂志，2021，27(18)：227-233.
［15］Yoshitake F，Itoh S，Narita H，et al.Interleukin-6 directly inhibits osteoclast differentiation by suppressing receptor activator of NF-kappaB signaling pathways［J］.J Biol Chem，2008，283(17)：11535-11540.
［16］赵净颖，段小花，王秋婷，等.动物骨代谢相关信号通路研究进展［J］.遗传，2020，42(10)：979-992.
［17］Fleisch H，Hofstetter W，Felix R，et al.The role of macrophage stimulating factor MCSF in bone resorption［J］.Osteoporosis Int，1993，3(Suppl 1)：108-110.
［18］Millan MM.The role of estrogen receptor in bone cells［J］.Clin Rev Bone Miner Metab，2015，13(2)：105-112.
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