中国骨质疏松杂志

miR-22-3p调控ROS/TXNIP/NLRP3通路对骨质疏松症小鼠破骨细胞活性的影响

Effect of miR-22-3p on osteoclast activity in osteoporotic mice by regulating the ROS/TXNIP/NLRP3 pathway

DOI：10.3969/j.issn.1006-7108.2025.08.010

中文关键词: 骨质疏松症破骨细胞活性氧硫氧还蛋白相互作用蛋白核苷酸结合寡聚化结构域样受体蛋白3

英文关键词:osteoporosis osteoclasts reactive oxygen species thioredoxin-interacting protein nucleotide binding oligomerization domain-like receptor protein 3

基金项目:

作者	单位
黄晓辉1 季泰令2*	1.山东第二医科大学，山东潍坊 252422 2.山东第二医科大学附属医院，山东潍坊 252422

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中文摘要:

目的基于ROS/TXNIP/NLRP3通路探讨miR-22-3p对骨质疏松症（osteoporosis，OP）小鼠破骨细胞活性的影响。方法雌性小鼠采用双侧卵巢摘除法构建OP模型，并随机分为OP组、agomir-NC组、miR-22-3p agomir组、miR-22-3p agomir+pc-NC组、miR-22-3p agomir+pc-TXNIP组，每组12只；另取12只小鼠为假手术（Sham）组。ELISA检测血清骨代谢指标（CTX-Ⅰ、TPACP5b、PINP、OC）和炎性因子（IL-1β、IL-18）含量；Micro-CT检测小鼠股骨骨小梁微结构变化（BMD、BV/TV、Tb.N、Tb.Th）；HE染色观察骨组织形态学变化；TRAP染色分析骨组织每平方毫米的破骨细胞数量（N.Oc/T.A）；二氢乙锭（DHE）染色检测骨组织ROS水平；qRT-PCR检测骨组织miR-22-3p、TXNIP和NLRP3的mRNA表达；Western blot检测骨组织TXNIP和NLRP3蛋白表达。结果与Sham组比较，OP组小鼠血清CTX-Ⅰ、TPACP5b、IL-1β、IL-18水平、股骨N.Oc/T.A、ROS水平、TXNIP与NLRP3的mRNA和蛋白表达升高，血清PINP、OC水平、股骨BMD、BV/TV、Tb.N、Tb.Th、miR-22-3p水平降低（P＜0.05）；与OP组、agomir-NC组比较，miR-22-3p agomir组小鼠血清CTX-Ⅰ、TPACP5b、IL-1β、IL-18水平、股骨N.Oc/T.A、ROS水平、TXNIP与NLRP3的mRNA和蛋白表达降低，血清PINP、OC水平、股骨BMD、BV/TV、Tb.N、Tb.Th、miR-22-3p水平升高（P＜0.05）；过表达TXNIP可明显消除miR-22-3p模拟物对OP小鼠骨代谢失衡的改善作用；miR-22-3p与TXNIP存在靶向关系。结论过表达miR-22-3p可能通过抑制ROS/TXNIP/NLRP3信号通路，抑制破骨细胞活性，进而改善OP小鼠的骨代谢失衡和骨微结构变化。

英文摘要:

Objective To investigate the effect of miR-22-3p on osteoclast activity in osteoporosis (OP) mice based on the ROS/TXNIP/NLRP3 pathway. Methods Female mice were used to construct the OP model by bilateral ovariectomy. Mice were randomly assigned into OP group, agomir-NC group, miR-22-3p agomir group, miR-22-3p agomir+pc-NC group, and miR-22-3p agomir+pc-TXNIP group, with 12 mice in each group. The other 12 mice were included as the sham surgery (Sham) group. ELISA was applied to detect serum bone metabolism indicators CTX-I, TPACP5b, PINP, OC, and inflammatory factors IL-1β and IL-18. Micro-CT was applied to detect microstructure changes in mouse femoral trabeculae (BMD, BV/TV, Tb.N, Tb.Th). HE staining was applied to observe morphological changes in bone tissue. TRAP staining was applied to analyze the number of osteoclasts per square millimeter of the bone tissue (N.Oc/T.A). Dihydroethidium (DHE) staining was applied to detect ROS level in the bone tissue. qRT-PCR was applied to detect the mRNA expressions of miR-22-3p, TXNIP, and NLRP3 in the bone tissue. Western blotting was applied to detect the expressions of TXNIP and NLRP3 proteins in the bone tissue. Results Compared to those in the Sham group, the levels of serum CTX-I, TPACP5b, IL-1β, IL-18, the levels of N.Oc/T.A and ROS in the femurs, and mRNA and protein expressions of TXNIP and NLRP3 in OP group increased, the levels of serum PINP, OC and the levels of BMD, BV/TV, Tb.N. Tb.Th and miR-22-3p in the femurs decreased (P<0.05). Compared to those in the OP group and agomir-NC group, the levels of serum CTX-I, TPACP5b, IL-1β, and IL-18, the levels of N.Oc/T.A and ROS in the femurs, and mRNA and protein expressions of TXNIP and NLRP3 in miR-22-3p agomir group decreased, and the levels of serum PINP and OC and the levels of BMD, BV/TV, Tb.N. Tb.Th, and miR-22-3p in the femurs increased (P<0.05). Overexpression of TXNIP greatly eliminated the improvement effect of miR-22-3p mimics on the bone metabolism imbalance in OP mice. MiR-22-3p had a targeted relationship with TXNIP. Conclusion Overexpression of miR-22-3p may relieve the bone metabolism imbalance and microstructure changes in OP mice by inhibiting the ROS/TXNIP/NLRP3 signaling pathway and by suppressing osteoclast activity.

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