首页|微小RNA-103a-3p通过肿瘤蛋白53调控凋亡抑制剂1/P53对骨质疏松症的影响

微小RNA-103a-3p通过肿瘤蛋白53调控凋亡抑制剂1/P53对骨质疏松症的影响

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目的 探讨微小RNA(miR)-103a-3p调控细胞肿瘤蛋白 53 调控凋亡抑制剂 1(TRIAP1)对成骨细胞分化以及去卵巢小鼠骨量的影响。方法 MC3T3-E1 细胞分为正常对照(NC)组、miR-103a-3p-NC组、miR-103a-3p模拟(mimc)组、miR-103a-3p mimic+TRIAP1-NC组、miR-103a-3p mimic+TRIAP1 mimic组。Real-time PCR检测细胞miR-103a-3p、TRIAP1、P53 的mRNA表达,MTT法和流式细胞术检测细胞增殖及凋亡,免疫荧光染色和茜红素染色检测细胞骨架F-actin表达和矿化情况,ELISA检测细胞碱性磷酸酶(ALP)活性。24 只雌性小鼠设为sham组、骨质疏松症(OP)组、miR-103a-3p antagonist-NC组和miR-103a-3p antagonist组,每组 6 只摘取双侧卵巢制备OP模型,sham组仅分离卵巢组织周围脂肪。测定骨组织miR-103a-3p、TRIAP1、P53、ALP、骨钙素(OCN)、骨桥蛋白(OPN)的mRNA表达,microCT测定骨密度(BMD)、骨矿物质含量(BMC),HE染色观察骨组织病理改变。结果 细胞转染miR-103a-3p mimic后,miR-103a-3p及P53 表达升高、TRIAP1 表达降低,细胞增殖降低、凋亡增加,F-actin表达减弱,钙结节数量减少,ALP活性降低(P<0。01);而在增加转染TRIAP1 mimic后,以上miR-103a-3p mimics导致的结果均得到显著逆转(P<0。01)。OP组小鼠骨组织miR-103a-3p、P53 表达升高,TRIAP1、ALP、OCN、OPN基因表达降低,BMD、BMC降低,骨组织结构破坏(P<0。05);miR-103a-3p antagonist组小鼠骨组织miR-103a-3p及P53 表达降低,TRIAP1、ALP、OCN、OPN基因表达升高,BMD、BMC升高,骨组织结构改善(P<0。05)。结论 MiR-103a-3p可介导TRIAP1/P53 抑制成骨细胞增殖及矿化,而miR-103a-3p拮抗治疗可减少OP小鼠骨量丢失。
Effects of microRNA-103a-3p on osteoporosis through tumor protein 53-regulated inhibitor of apoptosis 1/P53
Objective To investigate the efeects of microRNA(miR)-103a-3p regulates tumor protein 53-regulated inhibitor of apoptosis 1(TRIAP1)on osteoblast differentiation and bone mass in ovariectomized mice.Methods MC3T3-E1 cells were divided into normal group,miR-103a-3p-NC group,miR-103a-3p mimic group,miR-103a-3p mimic+TRIAP1-NC group,miR-103a-3p mimic+TRIAP1 mimic group.mRNA expression of miR-103a-3p,TRIAP1,P53 were detected by Real-time PCR;Cell proliferation and apoptosis were detected by MTT test and flow cytometry;cytoskeleton and mineralization of cells were detected by F-actin immunofluorescence staining and alizarin staining;alkaline phosphatase(ALP)activity was detected by ELISA.24 female mice were divided into sham group,osteoporosis(OP)group,miR-103a-3p antagonist-NC group,miR-103a-3p antagonist group(six in each group),extract bilateral ovaries to establish an OP model,sham group mice only isolated fat around ovarian tissue.mRNA expression of miR-103a-3p,TRIAP1,P53,ALP,osteocalcin(OCN),osteopontin(OPN)of bone tissue were detected;microCT detect bone mineral density(BMD),bone mineral content(BMC);haematoxylin eosin staining was used to observe pathological changes of bone tissue.Results After miR-103a-3p mimic was transfected into cells,the miR-103a-3p and P53 expression increased,TRIAP1 expression decreased,cell proliferation decreased,apoptosis increased,F-actin expression decreased,the number of calcium nodules decreased,and ALP enzyme activity decreased(P<0.01);however,after TRIAP1 mimic was additionally transfected into cells,the above result caused by miR-103a-3p mimics were significantly reversed(P<0.01).In OP group,the miR-103a-3p and P53 expression in bone tissue increased,the TRIAP1,ALP,OCN and OPN expression decreased,BMD and BMC were decreased,and bone tissue construct was damaged(P<0.05);in miR-103a-3p antagonist group,the miR-103a-3p and P53 expression in bone tissue decreased,TRIAP1,ALP,OCN,OPN expression increased,BMD and BMC increased,and bone tissue construct was improved(P<0.05).Conclusion MiRNA-103a-3p mediate TRIAP1/P53 to inhibit proliferation and mineralization of osteoblast,while miR-103a-3p antagonistic treatment reduce bone loss in OP mice.

OsteoporosisMircroRNA-103a-3pTumor protein 53-regulated inhibitor of apoptosis 1P53Osteogenic differentiationBone mineral densityReal-time PCRMouse

黄皆和、王茜、郏舜杰、杨晟

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浙江省台州市立医院骨科,浙江 台州 318099

浙江省台州市立医院肾内科,浙江 台州 318099

骨质疏松症 微小RNA-103a-3p 肿瘤蛋白 53 调控细胞凋亡抑制剂 1 P53 骨分化 骨密度 实时定量聚合酶链反应 小鼠

浙江省医药卫生科技计划

2022KY1392

2024

解剖学报
中国解剖学会

解剖学报

CSTPCD
影响因子:0.462
ISSN:0529-1356
年,卷(期):2024.55(2)
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