TNF-alpha mediates diabetes-enhanced chondrocyte apoptosis during fracture healing and stimulates chondrocyte apoptosis through FOXO1.
TNF-alpha mediates diabetes-enhanced chondrocyte apoptosis during fracture healing and stimulates chondrocyte apoptosis through FOXO1.
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DOI：
10.1002/jbmr.59
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发表时间：
2010-07
影响因子：
6.2
通讯作者：
Graves, Dana T.
中科院分区：
文献类型：
--
作者：
Kayal, Rayyan A.;Siqueira, Michelle;Alblowi, Jazia;McLean, Jody;Krothapalli, Nanarao;Faibish, Dan;Einhorn, Thomas A.;Gerstenfeld, Louis C.;Graves, Dana T.
关键词：
为了深入了解糖尿病对骨折愈合的影响，进行了实验，重点是从软骨到骨的过渡期间的软骨细胞凋亡。通过多次低剂量链脲佐菌素注射在小鼠中诱导1型糖尿病，并进行胫骨或股骨的简单横向骨折。进行大规模转录谱分析和基因集富集分析，以检查从第12天、第16天和第22天的骨折愈伤组织中分离的总RNA上的凋亡途径，这是软骨内骨形成的时期，此时软骨被再吸收，软骨细胞数量减少。ELISA法检测肿瘤坏死因子α（TNF-α）蛋白水平，Caspase-3活性测定。通过用TNF特异性抑制剂pegsunercept治疗小鼠来检查TNF的作用。体外研究调查了促凋亡转录因子FOXO 1在调节TNF诱导的软骨形成ATDC 5和C3 H10 T1/2细胞凋亡中的作用，ATDC 5和C3 H10 T1/2细胞是分化的软骨细胞的代表，这在软骨内骨化过程中很重要。mRNA分析显示，在糖尿病组中，当软骨吸收活跃时，在第16天与细胞凋亡相关的基因组上调，而不是第12天或第22天。这与TNF-α蛋白水平升高、软骨细胞凋亡、caspase-3活性增强和FOXO 1核转位增加相一致（p <0.05）。TNF的抑制显著降低了糖尿病小鼠的这些参数，但在血糖正常的对照小鼠中没有（p <0.05）。在体外使用siRNA沉默FOXO 1显着降低TNF诱导的分化软骨细胞凋亡和caspase活性。软骨细胞中FOXO 1的沉默显著降低了促凋亡基因caspase-3、caspase-8、caspase-9和TRAIL的mRNA水平。抑制caspase-8和caspase-9可显著减少TNF诱导的软骨细胞凋亡。这些结果表明，糖尿病引起的上调，在从软骨到骨的骨折愈合过程中的促凋亡基因。糖尿病通过一种机制增加软骨细胞凋亡，该机制涉及增加TNF-α的产生，其刺激软骨细胞凋亡并通过FOXO 1激活上调凋亡基因的mRNA水平。© 2010美国骨与矿物质研究学会。
To gain insight into the effect of diabetes on fracture healing, experiments were carried out focusing on chondrocyte apoptosis during the transition from cartilage to bone. Type 1 diabetes was induced in mice by multiple low-dose streptozotocin injections, and simple transverse fractures of the tibia or femur was carried out. Large-scale transcriptional profiling and gene set enrichment analysis were performed to examine apoptotic pathways on total RNA isolated from fracture calluses on days 12, 16, and 22, a period of endochondral bone formation when cartilage is resorbed and chondrocyte numbers decrease. Tumor necrosis factor α (TNF-α) protein levels were assessed by ELISA and caspase-3 by bioactivity assay. The role of TNF was examined by treating mice with the TNF-specific inhibitor pegsunercept. In vitro studies investigated the proapoptotic transcription factor FOXO1 in regulating TNF-induced apoptosis of chondrogenic ATDC5 and C3H10T1/2 cells as representative of differentiated chondrocytes, which are important during endochondral ossification. mRNA profiling revealed an upregulation of gene sets related to apoptosis in the diabetic group on day 16 when cartilage resorption is active but not day 12 or day 22. This coincided with elevated TNF-α protein levels, chondrocyte apoptosis, enhanced caspase-3 activity, and increased FOXO1 nuclear translocation (p < .05). Inhibition of TNF significantly reduced these parameters in the diabetic mice but not in normoglycemic control mice (p < .05). Silencing FOXO1 using siRNA in vitro significantly reduced TNF-induced apoptosis and caspase activity in differentiated chondrocytes. The mRNA levels of the proapoptotic genes caspase-3, caspase-8, caspase-9, and TRAIL were significantly reduced with silencing of FOXO1 in chondrocytic cells. Inhibiting caspase-8 and caspase-9 significantly reduced TNF-induced apoptosis in chondrogenic cells. These results suggest that diabetes causes an upregulation of proapoptotic genes during the transition from cartilage to bone in fracture healing. Diabetes increased chondrocyte apoptosis through a mechanism that involved enhanced production of TNF-α, which stimulates chondrocyte apoptosis and upregulates mRNA levels of apoptotic genes through FOXO1 activation. © 2010 American Society for Bone and Mineral Research.
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影响因子：
64.5
作者：
Brunet, A;Bonni, A;Greenberg, ME
通讯作者：
Greenberg, ME
FOXO转录因子直接激活BIM基因表达并促进交感神经元中的凋亡。
影响因子：
7.8
作者：
Gilley, Jonathan;Coffer, Paul J;Ham, Jonathan
通讯作者：
Ham, Jonathan
DOI：
10.1016/0922-3371(90)90079-c
发表时间：
1990-05-01
期刊：
CELL DIFFERENTIATION AND DEVELOPMENT
影响因子：
--
作者：
ATSUMI, T;MIWA, Y;IKAWA, Y
通讯作者：
IKAWA, Y
影响因子：
4
作者：
Huang, Haojie;Tindall, Donald J.
通讯作者：
Tindall, Donald J.
影响因子：
2.8
作者：
Aizawa, T;Kon, T;Gerstenfeld, LC
通讯作者：
Gerstenfeld, LC