基于绝经后女性股骨样本的骨髓造血干细胞自噬与骨质疏松相关性研究  

Correlation between autophagy of bone marrow hematopoietic stem cells and osteoporosis in femoral bone samples of postmenopausal women

作　　者：袁晔 王晓娟[2] 朱柯雨 魏祺 郑苗 邢丹[1] 徐又佳[4] 林剑浩[1] YUAN Ye;WANG Xiao-juan;ZHU Ke-yu;WEI Qi;ZHENG Miao;XING Dan;XU You-jia;LIN Jian-hao(Arthritis Clinical and Research Center,Peking University People's Hospital,Beijing 100044,China;Department of Hematology,First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710075,China;Osteoporosis Clinical Center,Second Affiliated Hospital of Soochow University,Suzhou 215004,Jiangsu,China;Department of Orthopedics,Second Affiliated Hospital of Soochow University,Suzhou 215004,Jiangsu,China)

机构地区：[1]北京大学人民医院骨关节科,北京100044 [2]西安交通大学第一附属医院血液内科,西安710075 [3]苏州大学附属第二医院骨质疏松症临床中心,江苏苏州215004 [4]苏州大学附属第二医院骨科,江苏苏州215004

出　　处：《中华骨质疏松和骨矿盐疾病杂志》2024年第3期221-229,共9页Chinese Journal Of Osteoporosis And Bone Mineral Research

基　　金：国家重点研发计划资助(2021YFC2501700、2021YFC2501702);国家自然科学基金(82372455、82272538);北京市自然科学基金-海淀原始创新联合基金(L222087);北京大学人民医院研究与发展基金资助项目(RDJP2023-21);江苏省医学重点实验室建设单位(JSDW202254)。

摘　　要：目的通过绝经后女性髋部骨折手术抽取分离获得骨髓造血干细胞,研究造血干细胞自噬与骨质疏松相关性。方法19例绝经后女性患者于髋部骨折手术中抽取骨髓,根据股骨颈、腰椎骨密度将患者分为骨质疏松组(n=13)和非骨质疏松组(n=6)。流式细胞术分选获得造血干细胞。成像流式细胞术检测两组造血干细胞自噬功能。反转录PCR(reverse transcription PCR,RT-PCR)检测两组骨髓造血干细胞自噬相关基因Atg7、LC3b、LAMP2a、P62、Atg5、Atg12表达。使用卵巢去势骨质疏松小鼠模型骨髓分选获得造血干细胞,RT-PCR检测造血干细胞自噬相关基因Atg7、LC3b、LAMP2a、P62、Atg5、Atg12表达。通过Western blot检测小鼠自噬相关蛋白表达。探究细胞自噬异常后血管内皮生长因子A(vascular endothelial growth factor A,VEGF-A)表达。结果骨质疏松患者骨髓造血干细胞自噬受抑制,自噬相关基因Atg7、LC3b、LAMP2a、P62、Atg5、Atg12表达下调(P<0.05)。小鼠骨质疏松组ATG7、BECLIN1、LC3-Ⅰ/LC3-Ⅱ蛋白表达受抑制。卵巢去势骨质疏松小鼠模型骨髓造血干细胞自噬相关基因Atg7、LC3b、LAMP2a、P62、Atg5、Atg12表达受抑制(P<0.05),细胞自噬被抑制后VEGF-A表达下降。结论骨髓造血干细胞自噬受抑制与骨质疏松有相关性。Objective To invetigate the correlation between autophagy of hematopoietic stem cells and osteoporosis in femoral bone samples of postmenopausal women obtained from hip fracture surgery.Methods Bone marrow of 19 postmenopausal women were collected from hip replacement surgery and the patients were divided into osteoporosis group(n=13)and non-osteoporosis group(n=6)according to the bone mineral density at femoral neck and lumbar spine.Hematopoietic stem cells were obtained by flow cytometry.Hematopoietic stem cells autophagy was measured by imaging flow cytometry.Reverse transcription PCR(RT-PCR)was performed to determine the expressions of autophagy related genes,including Atg7,LC3b,LAMP2a,P62,Atg5,Atg12.Ovariectomy osteoporosis mice model were established for collecting bone marrow hematopoietic stem cells.RT-PCR was performed to determine the expressions of autophagy related genes,including Atg7,LC3b,LAMP2a,P62,Atg5,Atg12.Autophagy related proteins were measured by western blot in mice.The expression of vascular endothelial growth factor A(VEGF-A)was detected when abnormal autophagy of bone marrow hematopoietic stem cells.Results Hematopoietic stem cells autophagy was inhibited in patients with osteoporosis and the expressions of autophagy related genes including Atg7,LC3b,LAMP2a,P62,Atg5,Atg12 were down-regulated(P<0.05).The expressions of ATG7,BECLIN1 and LC3-Ⅰ/LC3-Ⅱ were inhibited in osteoporotic mice.The expressions of autophagy related genes including Atg7,LC3b,LAMP2a,P62,Atg5,Atg12 were inhibited in ovariectomy osteoporosis mice and the expression of VEGF-A was reduced after the inhibition of the autophagy(P<0.05).Conclusion Inhibition of hematopoietic stem cells autophagy is related to osteoporosis.

关 键 词：髋部骨折 造血干细胞 骨质疏松 自噬 

分 类 号：R681[医药卫生—骨科学]