&nbsp;补体C3a受体在盲肠结扎穿孔致脓毒症大鼠认知障碍中的作用及机制

徐颖; 刘斌; 郑兴; 何宗钊

doi:10.16098/j.issn.0529-1356.2023.06.007

PDF(6273 KB)

解剖学报 ›› 2023, Vol. 54 ›› Issue (6) : 668-675. DOI: 10.16098/j.issn.0529-1356.2023.06.007

神经生物学

补体C3a受体在盲肠结扎穿孔致脓毒症大鼠认知障碍中的作用及机制

徐颖¹ 刘斌² 郑兴¹何宗钊^1*

作者信息 +

Role and mechanism of complement C3a receptor in the cognitive impairment of septic rats induced by cecal ligation and perforation

XU Ying¹ LIU Bin² ZHENG Xing¹ HE Zong-zhao^1*

Author information +

文章历史 +

摘要

目的探讨补体C3a受体（C3aR）在盲肠结扎穿孔（CLP）致脓毒症大鼠认知障碍中的作用及潜在的作用机制。方法 132只大鼠随机分为假手术组（sham组）和脓毒症组（CLP组），每组的初始数量为66只动物，每个时间点设置22只动物。构建SD大鼠CLP动物模型，于术后1、15和30 d取血清和脑（海马和前额叶皮质），采用ELISA法测定肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）和IL-6水平。Western blotting检测脑标本中Tau蛋白（Tau）、磷酸化Tau（p-Tau）和C3aR蛋白表达。将66只大鼠随机分为3组：sham组、CLP组和C3aR拮抗剂(C3aRa)干预组。于CLP后第15、17和19天，C3aRa干预大鼠于CLP术后第30天接受抑制性回避测试和物体识别测试，并采用免疫荧光法检测海马中C3aR、离子钙接头蛋白分子1（Iba1）、胶质纤维酸性蛋白（GFAP）和p-Tau表达。结果与sham组相比，CLP组大鼠血清及脑组织中TNFα、IL-1β和IL-6在CLP后30 d内先增加后减少。在CLP组海马组织和前额叶皮层中分别观察到术后30 d和术后1、15 d Tau磷酸化显著增强（P<0.05）。与sham组相比，CLP组术后15 d和30 d在大鼠海马和前额叶皮层中均观察到C3aR蛋白水平显著增加（P<0.05）。与CLP组相比，C3aRa干预后大鼠海马内源性C3aR含量明显降低（P<0.05），Iba1、GFAP和p-Tau免疫染色明显抑制（P<0.05）。C3aRa干预CLP大鼠可逆转认知功能损伤。结论 C3aR在脓毒症诱导神经退行性过程中相关生物化学和行为学变化发展中发挥重要作用。通过在海马注射C3aRa能够改善由脓毒症引发的神经退化过程。

Abstract

Objective To study the role of complement C3a receptor (C3aR) in cognitive impairment in rats with sepsis induced by cecal ligation and puncture (CLP), and to explore the possible mechanism. Methods Totally 132 rats were randomly divided into sham operation group (sham group) and sepsis group (CLP group). The initial number of each group was 66 animals, and 22 animals were set at each time point. The SD rat CLP animal model was constructed, and serum and brain (hippocampus and prefrontal cortex) samples were collected at day 1, day 15 and day 30 after operation. The levels of tumor necrosis factor-α(TNF-α), interleukin-1β (IL-1β) and IL-6 in the samples were determined by ELISA. Western blotting detected Tau protein (Tau), phosphorylated Tau（p-Tau）and C3aR expression in brain samples. Totally 66 rats were randomly divided into 3 groups, sham operation group, CLP group and C3aR antagonist(C3aRa) intervention group. On 15th, 17th, and 19th days after CLP, C3aRa intervened in rats, and they received inhibition avoidance test and object recognition test 30 days after CLP. The expressions of C3aR, lonized calcium binding adapter molecule 1(Iba1), GFAP and p-Tau in the hippocampus were detected by immunofluorescence. Results Compared with the sham group, the serum and brain tissue (TNFα, IL-1β and IL-6) of rats in the CLP group first increased and then decreased within 30 days after CLP. In the hippocampus and prefrontal cortex of CLP group, Tau phosphorylation was significantly enhanced at day 30 and day 1 and 15 after surgery, respectively (P<0.05). Compared with the sham group, C3aR protein levels in hippocampus and prefrontal cortex of rats in CLP group increased significantly at day 15 and 30 after operation (P<0.05). Compared with the CLP group, the endogenous C3aR content decreased significantly after C3aRa intervention (P<0.05), and Iba1, GFAP and p-Tau immunostaining were significantly inhibited (P<0.05). The C3aRa intervention in CLP rats reversed the cognitive impairment. Conclusion C3aR plays an important role in the development of biochemical and behavioral changes commonly associated with the onset of sepsis-induced neurodegenerative processes. C3aRa can be injected into the hippocampus to counteract the neurodegenerative process induced by sepsis.

导出引用

徐颖刘斌郑兴何宗钊.

补体C3a受体在盲肠结扎穿孔致脓毒症大鼠认知障碍中的作用及机制

[J]. 解剖学报. 2023, 54(6): 668-675 https://doi.org/10.16098/j.issn.0529-1356.2023.06.007

XU Ying LIU Bin ZHENG Xing HE Zong-zhao. Role and mechanism of complement C3a receptor in the cognitive impairment of septic rats induced by cecal ligation and perforation[J]. Acta Anatomica Sinica. 2023, 54(6): 668-675 https://doi.org/10.16098/j.issn.0529-1356.2023.06.007

中图分类号： R714

参考文献

［1］Hu RH，,Zheng YL，Cheng F, et al. Dvances in precision immunotherapeutic research of sepsis［J］. Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care,2019, 26(4):504-506.(in Chinese）

胡荣华, 郑颜磊, 程飞,等. 脓毒症精准免疫治疗的新进展［J］. 中国中西医结合急救杂志, 2019, 26(4):504-506.

［2］Ren C, Yao R, Zhang H, et al. Sepsis-associated encephalopathy: a vicious cycle of immunosuppression［J］. J Neuroinflammation, 2020, 17(1): 1-15.

［3］Chen H, Dong B, Shi Y, et al. Hydrogen alleviated neuronal injury and neuroinflammation induced by microglial activation via the Nrf2 pathway in sepsis-associated encephalopathy［J］. Neuroscience, 2021, 466(16): 87-100.

［4］Tao SY,You T,Bi YH,et al.The expression and purification of a new complement inhibitor complement receptor of the immunoglobulin superfamily［J］. Chinese Journal of Experimental Surgery, 2019, 36(7):1267-1269.(in Chinese）

陶思跃, 尤涛, 毕意辉,等. 补体免疫抑制剂-免疫球蛋白超家族补体受体的原核表达与纯化［J］. 中华实验外科杂志, 2019, 36(7):1267-1269.

［5］Pozo-Rodrigálvarez A, Ollaranta R, Skoog J, et al. Hyperactive Behavior and Altered Brain Morphology in Adult Complement C3a Receptor Deficient Mice［J］. Front Immunol, 2021, 12(3): 406.

［6］Shi MM, Zhang Y, Li J, et al. Effect of chronic restraint stress on the phenotypic transition of hippocampal astrocytes and depression-like behavior in mice［J］. Acta Anatomica Sinica, 2023, 54(1): 42-49. (in Chinese）

时萌萌, 张圆, 李菁, 等. 慢性束缚应激对小鼠海马星形胶质细胞表型转换及抑郁样行为的影响［J］. 解剖学报, 2023, 54(1): 42-49.

［7］Ma R, Hu JJ, Yao Y, et al. Role and mechanism study of complement C3/C3aR in okadaic acid-induced hyperphosphorylation of tau in SH-SY5Y cells［J］. Acta Neuropharmacologica,2017, 7(2): 35.(in Chinese）

马嵘, 胡珺捷, 姚艺, 等. 补体C3/C3aR在冈田酸诱导的SH-SY5Y细胞tau蛋白过度磷酸化中的作用及机制研究［J］. 神经药理学报, 2017, 7(2): 35.

［8］Zhang LY, Pan J, Mamtilahun M, et al. Microglia exacerbate white matter injury via complement C3/C3aR pathway after hypoperfusion［J］. Theranostics, 2020, 10(1): 74-90.

［9］Li J, Wang H, Du C, et al. hUC-MSCs ameliorated CUMS-induced depression by modulating complement C3 signaling-mediated microglial polarization during astrocyte-microglia crosstalk［J］. Brain Res Bull, 2020, 163(9): 109-119.

［10］Rocha M, Vieira A, Michels M, et al. Effects of S100B neutralization on the long-term cognitive impairment and neuroinflammatory response in an animal model of sepsis［J］. Neurochem Int, 2021, 142(4): 104906.

［11］Gao R, Ji MH, Gao DP, et al. Neuroinflammation-induced downregulation of hippocampacal neuregulin 1-ErbB4 signaling in the parvalbumin interneurons might contribute to cognitive impairment in a mouse model of sepsis-associated encephalopathy［J］. Inflammation, 2017, 40(2):387-400.

［12］Yu X, Xu H, Jiang JW, et al. Predictive value of SIRT2 in the progression of sepsis to persistent inflammation -immunosuppression and catabolism syndrome［J］. Chinese Journal of Emergency Medicine,2020, 29(4):541-546.(in Chinese）

余信, 许华, 蒋佳维,等. SIRT2对脓毒症进展为持续炎症-免疫抑制-分解代谢综合征的预测价值探讨［J］. 中华急诊医学杂志, 2020, 29(4):541-546.

［13］Zhe H, Zhao L, Ling Z, et al. Wip1 regulates blood-brain barrier function and neuro-inflammation induced by lipopolysaccharide via the sonic hedgehog signaling signaling pathway［J］. Mol Immunol, 2018, 93(9): 31-37.

［14］Zhang ZhB, Xu QP. Experimental Study of ginsenoside Rg1 combined with antibiotics in the treatment of acute lung injury in mice with sepsis［J］.Journal of Sichuan University(Medical Sciences),2020, 51(3): 371-375.(in Chinese）

张振波, 徐秋萍. 人参皂苷Rg1联合抗生素治疗小鼠脓毒症急性肺损伤［J］. 四川大学学报（医学版）, 2020, 51(3): 371-375.

［15］Zhang ZhY, Li Y, Zhang YH. Advances in the study of the nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome in Parkinson’s disease［J］.Chinese Journal of Neurology, 2020, 53(2):147-151.(in Chinese）

张展舆, 李彦, 张玉虎. 核苷酸结合寡聚化结构域样受体蛋白3炎性小体在帕金森病中的研究进展［J］. 中华神经科杂志, 2020, 53(2):147-151.

［16］Kimura T, Sharma G, Ishiguro K, et al. Phospho-tau bar code: analysis of phosphoisotypes of tau and its application to tauopathy［J］. Front Neurosci, 2018, 12(3): 44.

［17］Novochadlo M, Goldim MP, Bonfante S, et al. Folic acid alleviates the blood brain barrier permeability and oxidative stress and prevents cognitive decline in sepsis-surviving rats［J］. Microvasc Res, 2021, 137(12): 104193.

［18］Paouri E, Georgopoulos S. Systemic and CNS inflammation crosstalk: implications for Alzheimer’s disease［J］. Curr Alzheimer Res, 2019, 16(6): 559-574.

［19］Ahmad S, Bhatia K, Kindelin A, et al. The role of complement C3a receptor in stroke［J］. Neuromolecular Med, 2019, 21(4): 467-473.

［20］Song ShSh, Zhang Y, Zheng LH, et al.Role of C3a and C5a in focal segmental glomerulosclerosis［J］. Chinese Journal of Nephrology, 2019, 35(6):407-414.(in Chinese）

宋莎莎, 张颖, 郑林花,等. C3a和C5a在局灶性节段性肾小球硬化症中的作用［J］. 中华肾脏病杂志, 2019, 35(6):407-414.

［21］Li S, Li B, Zhang L, et al. A complement-microglial axis driving inhibitory synapse related protein loss might contribute to systemic inflammation-induced cognitive impairment［J］. Int Immunopharmacol, 2020, 87(6): 106814.

［22］Seidel G, Gaser C, G?tz T, et al. Accelerated brain ageing in sepsis survivors with cognitive long-term impairment［J］. Eur J Neurosci, 2020, 52(10): 4395-4402.