HMGB1 and TLR4 mediate skeletal muscle recovery in a murine model of hindlimb ischemia.
HMGB1 and TLR4 mediate skeletal muscle recovery in a murine model of hindlimb ischemia.
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DOI：
10.1016/j.jvs.2012.11.071
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发表时间：
2013-08
影响因子：
4.3
通讯作者：
Tzeng, Edith
中科院分区：
文献类型：
--
作者：
Sachdev, Ulka;Cui, Xiangdong;Tzeng, Edith
我们之前已经证明，危险信号高迁移率组框1(HMGB1)在给药于缺血肌肉时促进血管生成。HMGB1通过Toll样受体4(TLR4)和晚期糖基化终末产物受体(RAGE)传递信号。然而，这些受体在缺血性损伤和肌肉恢复中的作用尚不清楚。我们假设TLR4介导了组织修复和血管生成对缺血的反应。结扎对照组、TLR4基因敲除小鼠(C3H/HeOuJ)、无TLR4基因敲除小鼠(C3H/HeJ)以及RAGE基因敲除小鼠和C57B6对照小鼠。在其他实验中，对照组小鼠在股动脉结扎前预先给予抗HMGBI中和抗体。两周后，用激光多普勒血流成像(LDPI)评估肢体血流灌注，并报告缺血侧与非缺血侧肢体血流量的比率。对胫前肌的肌肉坏死、脂肪替代和血管密度进行组织学定量。体外培养的人真皮微血管内皮细胞(HDMVECs)在低氧条件下检测TLR4和RAGE的表达。用HMGB1单独处理和在抗TLR4抗体存在的情况下，检测磷酸化ERK(p-ERK)，这是一种对EC血管生成行为至关重要的信号分子。抗HMGB1抗体和TLR4信号缺陷均导致HEJ小鼠在股动脉结扎后两周出现明显的肌肉坏死。与TLR4失能的HeJ小鼠相比，对照组HeOuJ小鼠的坏死较少，但脂肪替代量较多。与对照C3H小鼠相比，对照C57B6小鼠表现出显著的肌肉再生，坏死很少。肌肉再生并不依赖于愤怒。虽然不同品系的血管密度没有差异，但与突变品系相比，RAGE和TLR4信号完整的小鼠缺血肢体的血流量较少。在体外，缺氧可使EC TLR4表达增加，而TLR4拮抗剂可降低HMGB1诱导的ERK活化。HMGB1和TLR4对肢体缺血后的肌肉坏死均有保护作用。然而，肌肉再生似乎与血管密度无关。HMGB1可能在体外激活EC的血管生成行为，这种激活可能受TLR4的调节。在缺乏TLR4和RAGE信号的小鼠中看到的血流改善可能表明这两种受体都有抗血管生成的作用，或者是TLR4和RAGE介导的炎症通路诱导的血管收缩。
We have previously shown that the danger signal High Mobility Group Box 1 (HMGB1) promotes angiogenesis when administered to ischemic muscle. HMGB1 signals through Toll-like receptor 4 (TLR4) as well as the receptor for advanced glycation end-products (RAGE). However, the actions of these receptors in ischemic injury and muscle recovery are not known. We hypothesize that TLR4 mediates tissue recovery and angiogenesis in response to ischemia. Femoral artery ligation was performed in control, TLR4 competent (C3H/HeOuJ), and incompetent (C3H/HeJ) mice, as well as RAGE knockout mice and their C57B6 control counterparts. In other experiments, control mice were pretreated with anti-HMGBI neutralizing antibody before femoral artery ligation. After two weeks, limb perfusion was evaluated using laser Doppler perfusion imaging (LDPI) and reported as the ratio of blood flow in the ischemic to nonischemic limb. Muscle necrosis, fat replacement, and vascular density in the anterior tibialis muscle were quantified histologically. In vitro, TLR4 and RAGE expression was evaluated in human dermal microvascular endothelial cells (HDMVECs) in response to hypoxia. HDMVECs treated with HMGB1 alone and in the presence of anti-TLR4 antibody were probed for phosphorylated ERK (p-ERK), a signaling molecule critical to EC angiogenic behavior. Both anti-HMGB1 antibody as well as defective TLR4 signaling in HeJ mice resulted in prominent muscle necrosis two weeks after femoral artery ligation. Control HeOuJ mice had less necrosis than TLR4 incompetent HeJ mice, but a greater amount of fat replacement. In contrast to control C3H mice, control C57B6 mice demonstrated prominent muscle regeneration with very little necrosis. Muscle regeneration was not dependent on RAGE. While vascular density did not differ between strains, mice with intact RAGE and TLR4 signaling had less blood flow in ischemic limbs compared to mutant strains. In vitro, EC TLR4 expression increased in response to hypoxia while TLR4 antagonism decreased HMGB1-induced activation of ERK. Both HMGB1 and TLR4 protect against muscle necrosis after hindlimb ischemia. However, muscle regeneration does not appear to be tied to vascular density. HMGB1 likely activates angiogenic behavior in EC in vitro, and this activation may be modulated by TLR4. The improvement in blood flow seen in mice with absent TLR4 and RAGE signaling may suggest anti-angiogenic roles for both receptors, or vasoconstriction induced by TLR4 and RAGE mediated inflammatory pathways.
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影响因子：
4.4
作者：
Peyssonnaux, Carole;Cejudo-Martin, Pilar;Nizet, Victor
通讯作者：
Nizet, Victor
影响因子：
10.8
作者：
Messina, LM;Brevetti, LS;Sarkar, R
通讯作者：
Sarkar, R
影响因子：
4.8
作者：
Rong, LL;Yan, SF;Schmidt, AM
通讯作者：
Schmidt, AM
影响因子：
2.7
作者：
Ngamkitidechakul, C;Twining, SS
通讯作者：
Twining, SS
DOI：
10.1083/jcb.152.6.1197
发表时间：
2001-03-19
期刊：
The Journal of cell biology
影响因子：
--
作者：
Degryse B;Bonaldi T;Scaffidi P;Müller S;Resnati M;Sanvito F;Arrigoni G;Bianchi ME
通讯作者：
Bianchi ME