[1]何智军△ 何伟 洪志楠 李子褀 魏秋实 陈晓俊 颜新昊.压电通道蛋白1调节激素性股骨头坏死大鼠的成骨细胞凋亡与血管生成[J].中国中医骨伤科杂志,2021,29(11):1-6.
　HE Zhijun HE Wei HONG Zhinan LI Ziqi WEI Qiushi CHEN Xiaojun YAN Xinhao.Piezo1 Regulates Osteoblast Apoptosis and Angiogenesis in Rats with Steroid-Induced Femoral Head Necrosis[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2021,29(11):1-6.
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压电通道蛋白1调节激素性股骨头坏死大鼠的成骨细胞凋亡与血管生成()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第29卷
期数:
2021年11期
页码:
1-6
栏目:
实验研究
出版日期:
2021-11-15

文章信息/Info

Title:
Piezo1 Regulates Osteoblast Apoptosis and Angiogenesis in Rats with Steroid-Induced Femoral Head Necrosis
文章编号:
1005-0205(2021)11-0001-06
作者:
何智军1; 2△ 何伟3 洪志楠3 李子褀3 魏秋实3 陈晓俊4 颜新昊4
Author(s):
HE Zhijun1; 2△ HE Wei3 HONG Zhinan3 LI Ziqi3 WEI Qiushi3 CHEN Xiaojun4 YAN Xinhao4
1Guangzhou University of Traditional Chinese Medicine, Guangzhou 510006,China; 2Department of the First Orthopedics, Panyu District Hospital of Traditional Chinese Medicine,Guangzhou 511400,China; 3Department of Arthrology, The Third Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangzhou 511400,China; 4First Clinical Medical College,Guangzhou University of Traditional Chinese Medicine Guangzhou 511400,China.
关键词:
压电通道蛋白1;  Yes相关蛋白1/β-连环蛋白信号通路;  激素性股骨头坏死;  凋亡;  血管生成
Keywords:
Piezo1;  Yap1/β-catenin signaling pathway;  hormone-induced femoral head necrosis;  apoptosis;  angiogenesis
分类号:
R-33
文献标志码:
A
摘要:
目的:探讨压电通道蛋白1(Piezo1)对激素性股骨头坏死(Osteonecrosis of the Femoral Head,ONFH)大鼠成骨细胞凋亡与血管生成的影响与机制 。方法:48只成年雄性Wistar大鼠(12周龄)分为4组(每组12只):类固醇激素(Steroid,STE)联合Piezo1沉默载体(siRNA-Piezo1,siP1)组(STE+siP1组),STE联合沉默载体阴性对照组(STE+siNC组),STE组及对照组。收集股骨头骨组织,TUNEL法检测凋亡率,三维微血管造影观察股骨头血管量的变化,qPCR检测Piezo1、凋亡标志物Bax、Caspase-3、Bcl-2的mRNA表达量,蛋白免疫印迹法检测Piezo1、血管生成标志物VEGFR2、CD3和Yap1/β-catenin信号蛋白的表达量。重组人YAP1蛋白(Rh-YAP1)及β-catenin激活剂(ICG-001)分别处理成骨细胞系(hFOB1.19)和脐静脉内皮细胞系(HUVEC)细胞。TUNEL法检测hFOB1.19的凋亡率,管状结构形成实验评估HUVEC的血管生成。结果:siP1成功抑制ONFH大鼠模型中股骨头组织Piezo1的表达,且siP1抑制股骨头组织的凋亡率和凋亡标志物的表达量,部分恢复ONFH模型中损失的股骨头血管量,并上调血管新生标志物水平。siP1抑制Yap1和β-catenin蛋白的表达。使用Rh-YAP1和ICG-001激活Yap1/β-catenin信号通路,导致hFOB1.19的凋亡明显增加,但是HUVEC细胞的血管新生能力降低。结论:ONFH模型中,沉默Piezo1通过抑制Yap1/β-catenin信号通路改善成骨细胞凋亡并促进血管生成。
Abstract:
Objective:To explore efficacy and mechanism of Piezo1, a mechanical sensitivity channel, on osteoblast apoptosis and angiogenesis in steroid-induced osteonecrosis of femoral head(ONFH)rats. Methods:48 adult male Wistar rats(12 weeks old)were divided into 4 groups(n=12), including steroid hormones(steroid, STE)combined with Piezo1 silencing carrier(siRNA-Piezo1, siP1)group(STE+siP1 group), STE combined with silence negative control group(STE+siNC group), STE group and control group. Femoral head osseous tissue was collected after treated. The apoptosis rate was detected by TUNEL method. The changes in amount of blood vessels in femoral head was observed by three-dimensional microangiography. The mRNA expression of Piezo1, apoptosis markers Bax, Caspase-3, and Bcl-2 were detected by qPCR method. Western Blot was used to detect the expression of Piezo1, angiogenesis markers VEGFR2, CD3 and Yap1/β-catenin signaling protein. The cells of human osteoblastic cell line(hFOB1.19)and umbilical vein endothelial cell line(HUVEC)were treated with recombinant human YAP1 protein(Rh-YAP1)and β-catenin activator(ICG-001)respectively. TUNEL method was used to detect apoptosis rate of hFOB1.19. Tube structure formation experiment was used to evaluate angiogenesis ability of HUVEC. Results:SiP1 successfully inhibited the expression of Piezo1 in femoral head tissue in ONFH rat model. It also inhibited the apoptotic rate of femoral head tissue and the expression of apoptosis markers, partially restored the loss of femoral head blood vessels in ONFH model, and up-regulated the angiogenesis Marker level. SiP1 inhibited the expression of Yap1 and β-catenin protein. Using Rh-YAP1 and ICG-001 to activate Yap1/β-catenin signaling pathway resulted in a significant increase in apoptosis of hFOB1.19, but the angiogenesis ability of HUVEC cells was decreased. Conclusion:In ONFH model, silencing Piezo1 can improve osteoblast apoptosis and promote angiogenesis by inhibiting Yap1/β-catenin signaling pathway.

参考文献/References:

[1] 雷志强,曾平,陈卫衡,等.股骨头坏死流行病学特点分析[J].中医正骨,2020,32(1):4-6.
[2] LEI P,DU W,LIU H,et al.Free vascularized iliac bone flap based on deep circumflex iliac vessels graft for the treatment of osteonecrosis of femoral head[J].J Orthop Surg Res,2019,14(1):397.
[3] PARPAITE T,COSTE B.Piezo channels[J].Curr Biol,2017,27(7):R250-R252.
[4] ZHOU T,GAO B,FAN Y,et al.Piezo1/2 mediate mechanotransduction essential for bone formation through concerted activation of NFAT-YAP1-?-catenin[J].Elife,2020,9:e52779.
[5] ZHUN W,DONGHAI L,ZHOUYUAN Y,et al.Efficiency of cell therapy to gc-induced ONFH:BMSCs with DKK-1 interference is not superior to unmodified BMSCs[J].Stem Cells Int,2018:1340252.
[6] ZHENG H,YANG E,PENG H,et al.Gastrodin prevents steroid-induced osteonecrosis of the femoral headin rats by anti-apoptosis[J].Chin Med J,2014,127(22):3926-3931.
[7] HUANG R,ZHAN Q,HU W,et al.Association of ABCB1 and CYP450 gene polymorphisms and their DNA methylation status with steroid-induced osteonecrosis of the femoral head in the Chinese population[J].Genet Test Mol Biomarkers,2020,24(12):789-797.
[8] SUN W,CHI S,LI Y,et al.The mechanosensitive Piezo1 channel is required for bone formation[J].Elife,2019,8:e47454.
[9] LI J,HOU B,TUMOVA S,et al.Piezo1 integration of vascular architecture with physiological force[J].Nature,2014,515(7526):279-282.
[10] WANG L,YOU X,LOTINUN S,et al.Mechanical sensing protein PIEZO1 regulates bone homeostasis via osteoblast-osteoclast crosstalk[J].Nat Commun,2020,11(1):282.
[11] PENG P,NIE Z,SUN F,et al.Glucocorticoids induce femoral head necrosis in rats through the ROS/JNK/c-Jun pathway[J].FEBS Open Bio,2021,11(1):312-321.
[12] MORLEY LC,SHI J,GAUNT H J,et al.Piezo1 channels are mechanosensors in human fetoplacental endothelial cells[J].Mol Hum Reprod,2018,24(10):510-520.
[13] CHEN J,MEI Z,HUANG B,et al.IL-6/YAP1/β-catenin signaling is involved in intervertebral disc degeneration[J].J Cell Physiol,2019,234(5):5964-5971.
[14] BENHAM-PYLE B W,PRUITT B L,NELSON W J.Cell adhesion.Mechanical strain induces E-cadherin-dependent Yap1 and β-catenin activation to drive cell cycle entry[J].Science,2015,348(6238):1024-1027.
[15] SUN Z,OU C,LIU J,et al.YAP1-induced MALAT1 promotes epithelial-mesenchymal transition and angiogenesis by sponging miR-126-5p in colorectal cancer[J].Oncogene,2019,38(14):2627-2644.
[16] VALLéE A,GUILLEVIN R,VALLéE J N.Vasculogenesis and angiogenesis initiation under normoxic conditions through Wnt/β-catenin pathway in gliomas[J].Rev Neurosci,2018,29(1):71-91.
[17] YU Z,FAN L,LI J,et al.Lithium prevents rat steroid-related osteonecrosis of the femoral head by β-catenin activation[J].Endocrine,2016,52(2):380-390.

备注/Memo

备注/Memo:
基金项目:国家自然科学基金项目(81873327)
1广州中医药大学(广州,510006)
2广州市番禺区中医院骨伤一科
2广州中医药大学第三附属医院关节科
3广州中医药大学第一临床医学院
△通信作者 E-mail:h413366714@163.com
更新日期/Last Update: 1900-01-01