摘要
目 的
趋化因子 (CX3CL1) 在神经性疼痛中起重要的生理病理作用, 然而其在糖尿病神经病理痛中的作用还有待研究。 本实验主要研究了在糖尿病小鼠痛阈下调的时间窗内, 脊髓背角中趋化因子 CX3CL1/趋化因子受体 (CX3CR1) 在触诱发痛发生与发展中的作用。
创新点
主要探讨 CX3CR1 在链脲佐菌素 (STZ) 诱导的 1 型糖尿病 (T1DM) 小鼠早期发生的机械痛性神经病变中的作用。
方 法
本实验采用健康雄性 C57BL/6 小鼠与 CX3CR1 KO 小鼠, 体重 20∼30 g, 隔夜禁食12 h (20 点至次日 8 点), 并连续三天腹腔注射 100 mg/kg 的 STZ 制备 T1DM 模型。 以空腹血糖浓度>11.1 mmol/L 且三周后小鼠机械痛阈值明显下降的情况视为 T1DM 模型制备成功。在小鼠机械痛阈下降的对 应时间点, 取腰段脊髓背角, 采用蛋白质印迹法 (western blot) 和免疫组化法测定 CX3CL1 及 CX3CR1 的表达情况。 同时, 在发生机械痛阈值下降的第三周时间鞘内给予 CX3CR1 的中和抗体, 进行机械刺激并观察其痛阈值的变化。
结 论
STZ 诱导的 T1DM 动物模型在早期表现为显著的 机械诱发痛, 并伴随脊髓背角 CX3CL1/CX3CR1 表达上调; 在痛阈下降期鞘内给予 CX3CR1 的中 和抗体可抑制糖尿病小鼠的痛行为。 与腹腔注射 STZ 形成 T1DM 的 C57BL/6 小鼠相比 CX3CR1 基因敲除的糖尿病小鼠机械痛阈值下降的时间 延迟, 程度减轻。 因此, 我们推测 CX3CL1/CX3CR1 可能参与 T1DM 机械痛的形成与发展。
References
Abbott CA, Malik RA, van Ross ERE, et al., 2011. Prevalence and characteristics of painful diabetic neuropathy in a large community-based diabetic population in the U.K. Diabetes Care, 34(10):2220–2224. https://doi.org/10.2337/dc11-1108
Badr G, Badr BM, Mahmoud MH, et al., 2012. Treatment of diabetic mice with undenatured whey protein accelerates the wound healing process by enhancing the expression of MTP-1α, MIP-2, KC, CX3CL1 and TGF-β in wounded tissue. BMC Immunol, 13:32. https://doi.org/10.1186/1471-2172-13-32
Callaghan BC, Cheng HT, Stables CL, et al., 2012. Diabetic neuropathy: clinical manifestations and current treatments. Lancet Neurol, 11(6):521–534. https://doi.org/10.1016/S1474-4422(12)70065-0
Cameron NE, Eaton SEM, Cotter MA, et al., 2001. Vascular factors and metabolic interactions in the pathogenesis of diabetic neuropathy. Diabetologia, 44(11):1973–1988. https://doi.org/10.1007/s001250100001
Cao H, Zhao YQ, 2008. Spinal glial activation contributes to pathological pain states. Neurosci Biobehav Rev, 32(5):972–983. https://doi.org/10.1016/j.neubiorev.2008.03.009
Clark AK, Yip PK, Malcangio M, 2009. The liberation of fractalkine in the dorsal horn requires microglial cathepsin S. J Neurosci, 29(21):6945–6954. https://doi.org/10.1523/JNEUROSCI.0828-09.2009
Dansereau MA, Gosselin RD, Pohl M, et al., 2008. Spinal CCL2 pronociceptive action is no longer effective in CCR2 receptor antagonist-treated rats. J Neurochem, 106(2):757–769. https://doi.org/10.1111/j.1471-4159.2008.05429.x
Dermanovic Dobrota V, Hrabac P, Skegro D, et al., 2014. The impact of neuropathic pain and other comorbidities on the quality of life in patients with diabetes. Health Qual Life Outcomes, 12:171. https://doi.org/10.1186/s12955-014-0171-7
Guo XX, Wang Y, Wang K, et al., 2018. Stability of a type 2 diabetes rat model induced by high-fat diet feeding with low-dose streptozotocin injection. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 19(7):559–569. https://doi.org/10.1631/jzus.B1700254
Inoguchi T, Li P, Umeda F, et al., 2000. High glucose level and free fatty acid stimulate reactive oxygen species production through protein kinase C-dependent activation of NAD(P)H oxidase in cultured vascular cells. Diabetes, 49(11):1939–1945. https://doi.org/10.2337/diabetes.49.11.1939
Ji RR, Chamessian A, Zhang YQ, 2016. Pain regulation by non-neuronal cells and inflammation. Science, 354(6312):572–577. https://doi.org/10.1126/science.aaf8924
Kiyomoto M, Shinoda M, Okada-Ogawa A, et al., 2013. Fractalkine signaling in microglia contributes to ectopic orofacial pain following trapezius muscle inflammation. J Neurosci, 33(18):7667–7680. https://doi.org/10.1523/JNEUROSCI.4968-12.2013
Lee YS, Morinaga H, Kim JJ, et al., 2013. The fractalkine/CX3CR1 system regulates β cell function and insulin secretion. Cell, 153(2):413–425. https://doi.org/10.1016/j.cell.2013.03.001
Lindia JA, McGowan E, Jochnowitz N, et al., 2005. Induction of CX3CL1 expression in astrocytes and CX3CR1 in microglia in the spinal cord of a rat model of neuropathic pain. J Pain, 6(7):434–438. https://doi.org/10.1016/j.jpain.2005.02.001
Mika J, Zychowska M, Popiolek-Barczyk K, et al., 2013. Importance of glial activation in neuropathic pain. Eur J Pharmacol, 716(1–3):106–119. https://doi.org/10.1016/j.ejphar.2013.01.072
Milligan E, Zapata V, Schoeniger D, et al., 2005. An initial investigation of spinal mechanisms underlying pain enhancement induced by fractalkine, a neuronally released chemokine. Eur J Neurosci, 22(11):2775–2782. https://doi.org/10.1111/j.1460-9568.2005.04470.x
Moore RA, Wiffen PJ, Derry S, et al., 2014. Gabapentin for chronic neuropathic pain and fibromyalgia in adults. Cochrane Database Syst Rev, (4):CD007938. https://doi.org/10.1002/14651858.CD007938.pub3
Nishikawa T, Edelstein D, Du XL, et al., 2000. Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage. Nature, 404(6779):787–790. https://doi.org/10.1038/35008121
Oyama T, Miyasita Y, Watanabe H, et al., 2006. The role of polyol pathway in high glucose-induced endothelial cell damages. Diabetes Res Clin Pract, 73(3):227–234. https://doi.org/10.1016/j.diabres.2006.02.010
Rajchgot T, Thomas SC, Wang JC, et al., 2019. Neurons and microglia; a sickly-sweet duo in diabetic pain neuropathy. Front Neurosci, 13:25. https://doi.org/10.3389/fnins.2019.00025
Souza GR, Talbot J, Lotufo CM, et al., 2013. Fractalkine mediates inflammatory pain through activation of satellite glial cells. Proc Natl Acad Sci USA, 110(27):11193–11198. https://doi.org/10.1073/pnas.1307445110
Sugimoto K, Yasujima M, Yagihashi S, 2008. Role of advanced glycation end products in diabetic neuropathy. Curr Pharm Des, 14(10):953–961. https://doi.org/10.2174/138161208784139774
Svensson CI, Fitzsimmons B, Azizi S, et al., 2005. Spinal p38β isoform mediates tissue injury-induced hyperalgesia and spinal sensitization. J Neurochem, 92(6):1508–1520. https://doi.org/10.1111/j.1471-4159.2004.02996.x
Tsuda M, Inoue K, Salter MW, 2005. Neuropathic pain and spinal microglia: a big problem from molecules in ‘small’ glia. Trends Neurosci, 28(2):101–107. https://doi.org/10.1016/j.tins.2004.12.002
Xu AK, Gong Z, He YZ, et al., 2019. Comprehensive therapeutics targeting the corticospinal tract following spinal cord injury. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 20(3):205–218. https://doi.org/10.1631/jzus.B1800280
Yao L, Wu YT, Tian GX, et al., 2017. Acrolein scavenger hydralazine prevents streptozotocin-induced painful diabetic neuropathy and spinal neuroinflammation in rats. Anat Rec (Hoboken), 300(10):1858–1864. https://doi.org/10.1002/ar.23618
Zhang Y, Yan J, Hu R, et al., 2015. Microglia are involved in pruritus induced by DNFB via the CX3CR1/p38 MAPK pathway. Cell Physiol Biochem, 35(3):1023–1033. https://doi.org/10.1159/000373929
Zhao H, Alam A, Chen Q, et al., 2017. The role of microglia in the pathobiology of neuropathic pain development: what do we know? Br J Anaesth, 118(4):504–516. https://doi.org/10.1093/bja/aex006
Zochodne DW, Verge VMK, Cheng C, et al., 2000. Nitric oxide synthase activity and expression in experimental diabetic neuropathy. J Neuropathol Exp Neurol, 59(9):798–807. https://doi.org/10.1093/jnen/59.9.798
Acknowledgments
We thank Prof. Zhi-qi ZHAO (Institutes of Brain Science, Fudan University, Shanghai, China) for his helpful criticism and advice to this work.
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Cheng-ming NI and Bing-yu LING were responsible for drafting the manuscript, and analysis and interpretation of data. Xiang XU collected and analyzed the data. He-ping SUN and Hui JIN contributed analysis of data and manuscript preparation. Yu-qiu ZHANG helped perform the analysis with constructive discussions. Hong CAO and Lan XU contributed the conception and design of the current study. All authors have read and approved the final manuscript. Therefore, all authors have full access to all the data in the study and take responsibility for the integrity and security of the data.
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Cheng-ming NI, Bing-yu LING, Xiang XU, He-ping SUN, Hui JIN, Yu-qiu ZHANG, Hong CAO, and Lan XU declare that they have no conflict of interest.
All institutional and national guidelines for the care and use of laboratory animals were followed. All experiments were approved by the Animal Care and Use Committee of Fudan University, Shanghai, China and followed the policies issued by the guidelines for pain research of the International Association for the Study of Pain (IASP).
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Project supported by the National Natural Science Foundation of China (Nos. 81771208 and 81971043), the Health and Family Planning Commission of Wuxi (No. YGZXM1406), the Wuxi Municipal Bureau on Science and Technology (No. CSE31N1614), the Fundamental Research Fund of Wuxi People’s Hospital (No. RKA201720), and the Technology for Social Development Project of Kunshan (No. KS1539), China
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Ni, Cm., Ling, By., Xu, X. et al. CX3CR1 contributes to streptozotocin-induced mechanical allodynia in the mouse spinal cord. J. Zhejiang Univ. Sci. B 21, 166–171 (2020). https://doi.org/10.1631/jzus.B1900439
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DOI: https://doi.org/10.1631/jzus.B1900439