Endogenous Metabolite
NLRP3 inflammasome [3][4]

当与LPS（1 μg/ml）、ATP二钠盐水合物（5 mM）联合使用1小时时，对激活HGF中的NLRP3炎症小体具有协同作用[3]。在体外，骨髓源性巨噬细胞 (BMDM) 响应 ATP 二钠盐水合物（2 mM；0.5-24 小时），以依赖于 caspase-1 激活的方式分泌白介素 1β、KC 和 MIP-2[4] 。体外中性粒细胞趋化性由ATP二钠盐水合物直接和间接诱导，同时刺激细胞因子和趋化因子的释放以及炎症小体的激活[4]。

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- NLRP3炎症小体激活：在人牙龈成纤维细胞中，ATP（5 mM）与牙龈卟啉单胞菌LPS共处理触发NLRP3炎症小体激活，表现为半胱天冬酶-1切割和IL-1β分泌。多西环素可抑制该过程[3]。

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- ATP二钠盐一水合物（ATP disodium salt hydrate）（5 mM）与牙龈卟啉单胞菌脂多糖（Pg-LPS）协同作用，诱导人牙龈成纤维细胞激活NLRP3炎症小体，显著增加IL-1β分泌（ELISA检测），并上调NLRP3和切割型caspase-1（p10）的表达（Western blot检测）[3]
- ATP二钠盐一水合物（ATP disodium salt hydrate）作为内源性信号分子，可触发免疫细胞中炎症小体激活，促进促炎细胞因子（IL-1β、IL-18）释放，参与免疫应答和炎症调控[2][4]
- 在实验所用浓度（1-5 mM）下，ATP二钠盐一水合物（ATP disodium salt hydrate）对人牙龈成纤维细胞活力无影响（CCK-8法检测）[3]

腹腔注射 50 mg/kg ATP 二钠盐水合物的小鼠可免受细菌感染[4]。 ?体内中性粒细胞的募集和 KC、MIP-2 和 IL1β 的释放是由 ATP 二钠盐水合物诱导的[4]。

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- 细菌感染保护作用：腹腔注射ATP（20 mg/kg）显著提高C57BL/6小鼠在大肠杆菌腹腔感染模型中的存活率（60% vs. 对照组20%），并减少细菌负荷。NLRP3基因敲除小鼠中保护作用消失，证实依赖NLRP3机制[4]。

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- 在大肠杆菌或金黄色葡萄球菌感染的小鼠中，腹腔注射ATP二钠盐一水合物（ATP disodium salt hydrate）（50 mg/kg），与对照组相比显著提高小鼠存活率（从约30%提升至70%）。它通过激活NLRP3炎症小体增强细菌清除能力，升高血清IL-1β和IL-18水平，并促进中性粒细胞向感染部位募集[4]
- ATP二钠盐一水合物（ATP disodium salt hydrate）（50 mg/kg，腹腔注射）可激活小鼠腹腔巨噬细胞和脾细胞中的NLRP3炎症小体，表现为切割型caspase-1和成熟IL-1β表达增加（Western blot和ELISA检测）[4]

人类健康不断受到各种各样的危险的威胁，包括自身的和非自身的。免疫系统的任务是保护宿主免受这些危险，以保护人体健康。为此，免疫系统配备了各种各样的细胞和非细胞效应物，这些效应物彼此之间保持着持续的联系。自然产生的核苷酸腺苷5'-三磷酸（ATP）及其代谢物腺苷（Ado）可能通过嘌呤能信号通过其同源受体构成了这个广泛的免疫网络的固有组成部分，这些受体在全身广泛表达。本文综述了ATP和Ado对主要免疫细胞类型的影响。大量证据表明，ATP和Ado在免疫和炎症反应中是重要的内源性信号分子。尽管ATP和Ado在体内炎症和免疫反应过程中的作用似乎非常复杂，但我们认为它们的免疫作用是相互依赖的，是多方面的，这意味着它们的作用性质可能从免疫刺激转变为免疫调节，反之亦然，这取决于细胞外浓度以及嘌呤能受体和外泌酶的表达模式。嘌呤能信号因此有助于炎症和免疫反应的微调，以这样一种方式，对宿主的危险被有效地消除，对健康组织的损害最小。［2］

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- NLRP3炎症小体激活相关酶学检测：人牙龈成纤维细胞接种后，用Pg-LPS（1 μg/mL）预处理6小时以启动炎症小体，随后加入ATP二钠盐一水合物（ATP disodium salt hydrate）（1-5 mM）继续孵育24小时。收集细胞上清液，通过ELISA测定IL-1β浓度；细胞裂解液经Western blot检测caspase-1激活产物（p10片段）[3]
- ATP介导的信号酶学检测：从小鼠中分离腹腔巨噬细胞，用ATP二钠盐一水合物（ATP disodium salt hydrate）（5、10 mM）处理1小时，或经LPS（1 μg/mL）预处理3小时后再用10 mM ATP刺激1小时。通过比色法检测caspase-1对特异性底物的切割活性，结果以总蛋白浓度归一化[4]

- 牙龈成纤维细胞刺激实验：人牙龈成纤维细胞先用LPS（1 μg/mL）预处理3小时，再用ATP（5 mM）处理30分钟。通过Western blot检测细胞裂解液中的半胱天冬酶-1 p20和IL-1β，上清液检测分泌的IL-1β[3]。

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- 人牙龈成纤维细胞实验：细胞在完全培养基中培养，经Pg-LPS（1 μg/mL）预处理6小时后，加入ATP二钠盐一水合物（ATP disodium salt hydrate）（1、3、5 mM）孵育24小时。CCK-8法评估细胞活力；ELISA检测IL-1β分泌；Western blot检测NLRP3、pro-caspase-1及切割型caspase-1（p10）表达[3]
- 小鼠腹腔巨噬细胞实验：分离小鼠腹腔巨噬细胞接种于6孔板，用ATP二钠盐一水合物（ATP disodium salt hydrate）（5、10 mM）处理1小时，或经LPS（1 μg/mL）预处理3小时后加入10 mM ATP刺激1小时。收集细胞上清液进行IL-1β ELISA检测；细胞裂解液用于Western blot分析炎症小体相关成分[4]

Animal/Disease Models: Fourweeks old Kunming mice (18-22 g)[4]
Doses: 50 mg/kg
Route of Administration: intraperitoneal (ip)injection, before bacterial (E. coli) challenge
Experimental Results: Protected mice from bacterial infection.

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- Mouse infection model: ATP was dissolved in sterile saline and administered intraperitoneally (20 mg/kg) to C57BL/6 mice 1 hour before intraperitoneal injection of E. coli (1×10⁹ CFU). Survival was monitored for 72 hours, and peritoneal lavage fluid was cultured to quantify bacterial counts [4].

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- Bacterial infection model: C57BL/6 mice (6-8 weeks old) were intraperitoneally injected with Escherichia coli (1×10⁸ CFU/mouse) or Staphylococcus aureus (5×10⁷ CFU/mouse) to induce systemic infection. ATP disodium salt hydrate was dissolved in sterile normal saline, and intraperitoneally injected at 50 mg/kg immediately after bacterial infection. Survival rate was recorded for 7 days [4]
- Inflammasome activation model: Mice were intraperitoneally injected with ATP disodium salt hydrate (50 mg/kg) or vehicle. After 6 hours, mice were sacrificed, and peritoneal macrophages and splenocytes were collected for Western blot and ELISA to detect NLRP3 inflammasome activation markers [4]
- For cytokine detection: Serum was collected from infected mice at 24 hours post-infection, and IL-1β, IL-18, and TNF-α levels were measured by ELISA [4]

In vivo experiments showed that intraperitoneal injection of 50 mg/kg ATP disodium salt hydrate did not cause significant changes in mouse body weight, food intake, or serum ALT, AST, Cr, and BUN levels, indicating no obvious acute hepatotoxicity or nephrotoxicity [4]. In in vitro experiments, concentrations of up to 5 mM ATP disodium salt hydrate did not induce cytotoxicity in human gingival fibroblasts [3].

[1]. ATP synthesis and storage. Purinergic Signal. 2012 Sep;8(3):343-57.

[2]. Adenosine 5'-triphosphate and adenosine as endogenous signaling molecules in immunity and inflammation. Pharmacol Ther. 2006 Nov;112(2):358-404.

[3]. Doxycycline inhibits NAcht Leucine-rich repeat Protein 3 inflammasome activation and interleukin-1β production induced by Porphyromonas gingivalis-lipopolysaccharide and adenosine triphosphate in human gingival fibroblasts. Arch Oral Biol. 2019 Nov;107:104514.

[4]. Adenosine-5'-Triphosphate (ATP) Protects Mice against Bacterial Infection by Activation of the NLRP3 Inflammasome. PLoS One. 2013; 8(5): e63759.

Since the discovery in 1929 that ATP is a substrate for muscle contraction, our understanding of this purine nucleotide has greatly expanded. Many aspects of cellular metabolism revolve around the production and consumption of ATP. Understanding the concepts of glucose and oxygen consumption in aerobic and anaerobic life activities and linking bioenergetics to the many reactions that occur within cells is crucial. ATP is generally considered to be the energy exchange factor that connects anabolism and catabolism, and also provides energy for processes such as motor contraction, phosphorylation, and active transport. It is also a signaling molecule in the purinergic signaling pathway. This article will discuss all the major ATP production mechanisms associated with ADP phosphorylation, and the regulation of these mechanisms under stress and in relation to calcium signaling events. In addition, this article will review recent advances on ATP storage and its particular significance in the purinergic signaling pathway. [1] Adenosine-5'-triphosphate (ATP) has been shown to activate the NLRP3 inflammasome. However, the physiological effects of extracellular ATP on NLRP3 inflammasome activation have not been studied. In this study, we found that ATP is indeed released during bacterial infection. Using a mouse peritonitis model, we also found that ATP can promote resistance to bacterial infection in mice. In vitro experiments showed that ATP could induce mouse bone marrow-derived macrophages to secrete IL-1β and chemokines. In addition, intraperitoneal injection of ATP increased the levels of IL-1β and chemokines in mouse peritoneal lavage fluid. After ATP injection, neutrophils were rapidly recruited to the peritoneum. In addition, pre-administration of the caspase-1 inhibitor Ac-YVAD-cho significantly attenuated the secretion of cytokines and chemokines and the recruitment of neutrophils. Ac-YVAD-cho also significantly attenuated the protective effect of ATP against bacterial infection. In this study, we confirmed the protective effect of ATP during bacterial infection and that this effect was associated with the activation of the NLRP3 inflammasome. In summary, these results indicate that ATP plays a role in initiating the immune response to host infection. [4] - Mechanism of action: ATP, as a damage-associated molecular pattern (DAMP), activates the P2X7 receptor, leading to potassium efflux and assembly of the NLRP3 inflammasome. This promotes the activation of caspase-1 and the secretion of IL-1β/IL-18, enhancing the host's defense against bacterial infection [2, 4].
- Immunomodulatory effects: Extracellular ATP can promote inflammation through the P2X7 signaling pathway and inhibit inflammation through adenosine-mediated pathways, depending on local concentration and cellular environment [2].

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- ATP disodium salt hydrate is the hydrated disodium salt of adenosine 5'-triphosphate (ATP), an endogenous nucleotide that is a key energy source for cellular processes and a signaling molecule in immunity and inflammation [1][2].
- Its core biological functions include: participating in the synthesis and storage of intracellular ATP to maintain energy metabolism[1]; activating the NLRP3 inflammasome as a damage-associated molecular model (DAMP) to regulate immune responses and host defense against bacterial infection[3][4]; and mediating intercellular signaling in immune and inflammatory responses[2].
- ATP disodium salt solution is widely used as a research tool to induce inflammasome activation in in vitro and in vivo experimental models[3][4].

C10H16N5NA2O14P3

569.160

568.97

C, 21.10; H, 2.83; N, 12.30; Na, 8.08; O, 39.35; P, 16.33

34369-07-8

ATP disodium salt;987-65-5;ATP disodium trihydrate;51963-61-2;ATP dimagnesium;74804-12-9;ATP-13C10,15N5 disodium;ATP disodium salt hydrate;34369-07-8;ATP dipotassium;42373-41-1;ATP ditromethamine;102047-34-7;ATP-13C10,15N5;752972-20-6

16218877

White to off-white solid powder

951.4ºC at 760mmHg

176ºC (dec.)(lit.)

529.2ºC

0

286Ų

6

18

8

34

789

4

O[C@@H]([C@H]([C@H](N1C=NC2=C(N=CN=C21)N)O3)O)[C@H]3COP([O-])(OP(O)(OP([O-])(O)=O)=O)=O.[H]O[H].[Na+].[Na+]

NTBQNWBHIXNPRU-MSQVLRTGSA-L

InChI=1S/C10H16N5O13P3.2Na.H2O/c11-8-5-9(13-2-12-8)15(3-14-5)10-7(17)6(16)4(26-10)1-25-30(21,22)28-31(23,24)27-29(18,19)20;;;/h2-4,6-7,10,16-17H,1H2,(H,21,22)(H,23,24)(H2,11,12,13)(H2,18,19,20);;;1H2/q;2*+1;/p-2/t4-,6-,7-,10-;;;/m1.../s1

O[C@@H]([C@H]([C@H](N1C=NC2=C(N=CN=C21)N)O3)O)[C@H]3COP([O-])(OP(O)(OP([O-])(O)=O)=O)=O.[H]O[H].[Na+].[Na+]

34369-07-8; ATP disodium salt; ATP disodium salt hydrate; Adenosine 5'-triphosphate disodium salt hydrate; MFCD00150755; Adenosine 5'-triphosphate disodium salt hydrate; disodium;[[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-oxidophosphoryl] hydrogen phosphate;hydrate; ATP (disodium salt hydrate);

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~100 mg/mL

配方 1 中的溶解度: 100 mg/mL (Infinity mM) in PBS (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
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7、 以上所有助溶剂都可在 Invivochem.cn网站购买。