The mechanism of Annexin A1 to modulate TRPV1 and nociception in dorsal root ganglion neurons.
The mechanism of Annexin A1 to modulate TRPV1 and nociception in dorsal root ganglion neurons.
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膜联蛋白 A1 调节背根神经节神经元 TRPV1 和伤害感受的机制
DOI：
10.1186/s13578-021-00679-1
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发表时间：
2021-08-26
影响因子：
7.5
通讯作者：
Pei L
中科院分区：
文献类型：
--
作者：
Zhang Y;Ma S;Ke X;Yi Y;Yu H;Yu D;Li Q;Shang Y;Lu Y;Pei L
膜联蛋白A1（ANXA 1）通过ANXA 1受体甲酰肽受体2（FPR 2/ALX（脂氧素A4受体），FPR 2）在背根神经节（DRG）水平发挥抗伤害性作用。然而，其机制仍有待阐明。通过使用辐射热、热/冷板、甩尾、von Frey和Randall-Selitto测试来检测完整和化学的伤害性感受，（辣椒素、薄荷脑、芥末油、福尔马林或CFA）注射AnxA 1条件性敲除（AnxA 1-/-）小鼠，在培养的DRG神经元中应用钙成像和膜片钳记录来测量神经元兴奋性，进行免疫荧光和蛋白质印迹来检测TRPV 1的蛋白水平，FPR 2及其下游分子，通过免疫荧光和免疫共沉淀技术研究钙调素（CaM）与TRPV 1的相互作用，旨在揭示ANXA 1抗伤害作用的分子和细胞机制。AnxA 1 −/−小鼠对有害热表现出显著的敏感性（平均值± SD，Hargreaves试验中6.2 ± 1.0 s vs.9.9 ± 1.6 s;热板试验中13.6 ± 1.5 s vs.19.0 ± 1.9 s; n = 8; P < 0.001），辣椒素（101.0 ± 15.3 vs. 76.2 ± 10.9; n = 8; P < 0.01），福尔马林（早期：169.5 ± 32.8 s vs76.0 ± 21.9 s; n = 8; P < 0.05;晚期：444.6 ± 40.1 s vs320.4 ± 33.6 s; n = 8; P < 0.01）和CFA（3.5 ± 0.8s对5.9 ± 1.4s; n = 8; P < 0.01）。此外，我们发现辣椒素诱导的Ca ~（2+）反应，TRPV 1电流和神经元放电在AnxA 1缺陷的DRG神经元显着增加。此外，ANXA 1模拟肽Ac 2 -26可显著增加细胞内Ca 2+，抑制TRPV 1电流，激活PLCβ，促进CaM-TRPV 1相互作用。Ac 2 -26的上述作用可被FPR 2拮抗剂Boc 2所减弱。选择性缺失AnxA 1可增强TRPV 1的敏感性，降低热刺激或辣椒素刺激的伤害性感受，而模拟肽Ac 2 -26可通过FPR 2和下游PLCβ-Ca 2 +-CaM信号降低TRPV 1的敏感性。本研究为开发新的炎性疼痛镇痛药物提供了可能的靶点。在线版本包含补充材料，可通过10.1186/s13578-021-00679-1获得。
Annexin A1 (ANXA1) exerts anti-nociceptive effect through ANXA1 receptor formyl peptide receptor 2 (FPR2/ALX (receptor for lipoxin A4), FPR2) at the dorsal root ganglia (DRG) level. However, the mechanisms remain elucidated. By using radiant heat, hot/cold plate, tail flick, von Frey, and Randall-Selitto tests to detect nociception in intact and chemical (capsaicin, menthol, mustard oil, formalin or CFA) injected AnxA1 conditional knockout (AnxA1−/−) mice, applying calcium imaging and patch clamp recordings in cultured DRG neurons to measure neuronal excitability, conducting immunofluorescence and western blotting to detect the protein levels of TRPV1, FPR2 and its downstream molecules, and performing double immunofluorescence and co-immunoprecipitation to investigate the interaction between Calmodulin (CaM) and TRPV1; we aim to uncover the molecular and cellular mechanisms of ANXA1’s role in antinociception. AnxA1−/− mice exhibited significant sensitivity to noxious heat (mean ± SD, 6.2 ± 1.0 s vs. 9.9 ± 1.6 s in Hargreaves test; 13.6 ± 1.5 s vs. 19.0 ± 1.9 s in hot plate test; n = 8; P < 0.001), capsaicin (101.0 ± 15.3 vs. 76.2 ± 10.9; n = 8; P < 0.01), formalin (early phase: 169.5 ± 32.8 s vs. 76.0 ± 21.9 s; n = 8; P < 0.05; late phase: 444.6 ± 40.1 s vs. 320.4 ± 33.6 s; n = 8; P < 0.01) and CFA (3.5 ± 0.8 s vs. 5.9 ± 1.4 s; n = 8; P < 0.01). In addition, we found significantly increased capsaicin induced Ca2+ response, TRPV1 currents and neuronal firing in AnxA1 deficient DRG neurons. Furthermore, ANXA1 mimic peptide Ac2-26 robustly increased intracellular Ca2+, inhibited TRPV1 current, activated PLCβ and promoted CaM-TRPV1 interaction. And these effects of Ac2-26 could be attenuated by FPR2 antagonist Boc2. Selective deletion of AnxA1 in DRG neurons enhances TRPV1 sensitivity and deteriorates noxious heat or capsaicin induced nociception, while ANXA1 mimic peptide Ac2-26 desensitizes TRPV1 via FPR2 and the downstream PLCβ-Ca2+-CaM signal. This study may provide possible target for developing new analgesic drugs in inflammatory pain. The online version contains supplementary material available at 10.1186/s13578-021-00679-1.
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