铁死亡途径探究骨碎补对去卵巢骨质疏松症大鼠的骨保护机制
Exploration of the bone protection mechanism of rhizoma drynariae through ferroptosis pathway in ovariectomized osteoporotic rats
  
DOI:10.3969/j.issn.1006-7108.2025.09.003
中文关键词:  骨碎补  绝经后骨质疏松症  去卵巢绝经后骨质疏松症大鼠模型  铁死亡
英文关键词:rhizoma drynariae  postmenopausal osteoporosis  ovariectomized postmenopausal osteoporosis rat model  ferroptosis
基金项目:国家自然科学基金资助项目(8217151988)
作者单位
齐朋朋1.2 陈丹3 房义辉3 李念虎1 徐展望1 薛海鹏1* 李霞4 1.山东中医药大学附属医院山东 济南 250014 2.山东中医药大学山东 济南250355 3.聊城市中医医院山东聊城252000 4.山东中医药大学中医药创新研究院山东 济南 250355 
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中文摘要:
      目的 通过动物实验验证基于铁死亡途径骨碎补是否对去卵巢骨质疏松模型大鼠具有骨保护作用以及机制探究。方法 30只SD大鼠随机平分为假手术组(SHAM)、模型组(OVX)、骨碎补组(OVX-DR)。模型组及骨碎补组构建去卵巢骨质疏松症大鼠模型,骨碎补组给予骨碎补水煎剂灌胃,剩余两组给予等剂量生理盐水灌胃处理,12周后取材检测。Micro-CT检测股骨骨微细结构;HE染色观察组织形态学改变;免疫荧光观察FTH1、SLC7A11、GPX4的表达;qRT-PCR法检测大鼠血液及骨组织FTH1、SLC7A11、GPX4、OPG、RUNX2、RANKL的mRNA表达;Western Blot检测各组大鼠骨组织FTH1、SLC7A11、GPX4、OPG、RUNX2、RANKL蛋白表达。结果 ①通过Micro-CT检测得出模型组的大鼠骨量明显低于假手术组,骨碎补组骨量高于模型组但低于假手术组。②HE染色显示模型组骨小梁较稀疏,骨碎补组骨小梁密度高于模型组但低于假手术组。③免疫荧光显示骨碎补组铁死亡相关指标FTH1、SLC7A11、GPX4表达高于模型组但低于假手术组,骨碎补可以降低FTH1、SLC7A11、GPX4的相对表达(P<0.05)。④qRT-PCR检测各组铁死亡及成破骨相关指标的mRNA表达水平结果:与模型组相对照,骨碎补组的OPG、RUNX2、FTH1、SLC7A11及GPX4的mRNA表达水平升高(P<0.05),RANKL的mRNA表达降低(P<0.05):与假手术组相比骨碎补组的OPG、RUNX2、FTH1、SLC7A11及GPX4的mRNA表达水平升高(P<0.05),RANKL的mRNA表达降低(P<0.05)。⑤ WB检测各组铁死亡及成破骨相关指标的蛋白水平结果:与模型组相对照,骨碎补组的OPG、RUNX2、FTH1、SLC7A11及GPX4的蛋白水平升高(P<0.05),RANKL的蛋白水平表达降低(P<0.05):与假手术组相比骨碎补组的OPG、RUNX2、FTH1、SLC7A11及GPX4的蛋白表达水平升高(P<0.05),RANKL的蛋白水平表达降低(P<0.05)。结论 骨碎补减少去卵巢绝经后骨质疏松症模型大鼠骨量丢失,防治绝经后骨质疏松的发生,其机制可能与铁死亡有关。
英文摘要:
      Objective Animal experiments were conducted to explore the bone protection mechanism of rhizoma drynariae through ferroptosis pathway in ovariectomized osteoporotic rats. Methods Thirty SD rats were randomly divided into SHAM group, OVX group, and OVX-DR group. The rat model of ovaricectomy osteoporosis was constructed in the model group and the rhizoma drynariae group. Rats in the rhizoma drynariae group received bone crushing hydration decoction by intragastric administration. Rats in the remaining two groups received equal dose of normal saline. Micro-CT was used to detect the microstructures of the femoral bone. HE staining was used to observe changes in tissue morphology, Immunofluorescence was used to observe the expression of FTH1, SLC7A11, and GPX4. qRT-PCR was used to detect the mRNA expressions of FTH1, SLC7A11, GPX4, OPG, RUNX2, and RANKL in the blood and bone tissue of rats. Western blotting was used to detect the protein expressions of FTH1, SLC7A11, GPX4, OPG, RUNX2, and RANKL in rat bone tissue. Results ①Micro-CT showed that the bone mass in the model group was significantly lower than that in the sham operation group, and the bone mass in the rhizoma drynariae group was higher than that in the model group but lower than that inthe sham operation group.②HE staining results showed that the bone trabecular density in the model group was sparse, and the bone trabecular density in the rhizoma drynariae group was higher than that in the model group but lower than that in the sham operation group.③ Immunofluorescence results showed that the expressions of FTH1, SLC7A11, and GPX4 related to ferroptosis in the rhizoma drynariae group were higher than those in the model group but lower than those in the sham operation group, and the relative expressions of FTH1, SLC7A11, and GPX4 in the rhizoma drynariae group decreased (P<0.05). ④ mRNA expression levels of ferroptosis and osteoclasts in each group were detected using qRT-PCR. The results showed that compared with the model group, the mRNA expression levels of OPG, RUNX2, FTH1, SLC7A11 and GPX4 in the rhizoma drynariae group increased (P<0.05), while the mRNA expression of RANKL decreased (P < 0.05). Compared with the sham operation group, the mRNA expression levels of OPG, RUNX2, FTH1, SLC7A11, and GPX4 increased (P < 0.05), and the mRNA expression of RANKL decreased (P < 0.05). ⑤Results of WB detection of the protein levels of ferroptosis and osteoclast-related indicators in each group showed that compared with the model group, the protein levels of OPG, RUNX2, FTH1, SLC7A11, and GPX4 in the rhizoma drynariae group increased (P<0.05), while the protein level of RANKL decreased (P < 0.05). Compared with the sham operation group, the protein expression levels of OPG, RUNX2, FTH1, SLC7A11, and GPX4 increased (P<0.05), and the protein level of RANKL decreased (P<0.05). Conclusion Rhizoma drynariae reduces bone loss and prevents the occurrence of postmenopausal osteoporosis in ovariectomized rats, and its mechanism may be related to ferroptosis.
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