脾切除术通过GTS-21诱导的胆碱能抗炎途径对脂多糖诱导的急性肾损伤的减轻作用。

Effect of splenectomy on attenuation of LPS-induced AKI through GTS-21-induced cholinergic anti-inflammatory pathway.

作者信息

Gao Yang, Kang Kai, Zhang Xinyu, Han Qiuyuan, Liu Haitao, Kong Weilan, Zhang Xing, Huang Rui, Yang Zhenyu, Qi Zhidong, Zheng Junbo, Li Ming, Li Jiayu, Liu Ruijin, Liu Yansong, Wang Sicong, Zhang Weihua, Wang Hongliang, Yu Kaijiang

机构信息

Department of Critical Care Medicine, The Second Affiliated Hospital of Harbin Medical University Harbin 150086, China.

Department of Critical Care Medicine, The First Affiliated Hospital of Harbin Medical University Harbin 150001, China.

出版信息

Am J Transl Res. 2019 Apr 15;11(4):2540-2549. eCollection 2019.

PMID:31105861

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6511767/

Abstract

This work was undertaken to explore the role of splenectomy on attenuation of lipopolysaccharide (LPS)-induced acute kidney injury (AKI) through GTS-21-induced cholinergic anti-inflammatory pathway. C57BL/6 mice were used to construct models of sepsis-induced renal injury. HE, Tunel and blood assays were used to determine the success of the model. The animals were examined after splenectomy with or without LPS and GTS-21+LPS treatments. The pathological changes and apoptosis in the renal tissue were detected using HE and Tunel assays. The contents of creatinine (Cr) and cystatin-C (Cys-C) were measured using ELISA. The expression of IL-6, NF-kB p65, Caspase-3, anti-apoptotic protein Bcl-2, apoptotic protein Bax and α7nAChR was quantified using qRT-PCR. The expression of Bcl-2, Bax, Caspase-3, IL-6, NF-kB p65, α7nAChR and p-STAT3 was using assessed using Western blot analysis. HE, Tunel, BUN and serum creatinine (SC) assay showed that renal injury models were successfully established. Compared with the control, the apoptosis in the LPS group was significantly increased and decreased after GTS-21 treatment. However, splenectomy combined with GTS-21 increased the apoptosis, indicating that splenectomy could partially offset the anti-apoptosis effect of GTS-21. In animals treated with LPS, the contents of Cr and Cys-C increased significantly. These contents reduced following GTS-21 treatment, but increased after splenectomy. After LPS treatment, the expression of IL-6, NF-kB p65, p-STAT3, Caspase-3 and Bax was significantly up-regulated, while the expression of α7nAChR and Bcl-2 significantly down-regulated. Compared with LPS treated mice, splenectomy reduced the expression of IL-6, NF-kB p65 and p-STAT3, suggesting that splenectomy inhibits the activation of α7nAChR pathway by the GTS-21. It is clear that GTS-21 effectively attenuates LPS-induced renal injury; splenectomy suppresses the anti-inflammatory and anti-apoptosis activity and renal protective effect of GTS-21. On other hand, splenectomy reduces the production of inflammatory cytokines in the circulation, and has certain protective effect on the kidney. Therefore, the impact of splenectomy on LPS-induced AKI depends on the strength of the two aspects.

摘要

本研究旨在探讨脾切除术通过GTS-21诱导的胆碱能抗炎途径对脂多糖（LPS）诱导的急性肾损伤（AKI）的缓解作用。采用C57BL/6小鼠构建脓毒症诱导的肾损伤模型。通过苏木精-伊红（HE）染色、TUNEL检测和血液检测来确定模型是否成功构建。对动物进行脾切除术后，分别给予或不给予LPS以及GTS-21+LPS处理，之后对动物进行检查。采用HE染色和TUNEL检测来检测肾组织中的病理变化和细胞凋亡情况。使用酶联免疫吸附测定（ELISA）法检测肌酐（Cr）和胱抑素C（Cys-C）的含量。采用实时定量聚合酶链反应（qRT-PCR）对白细胞介素-6（IL-6）、核因子-κB p65（NF-κB p65）、半胱天冬酶-3（Caspase-3）、抗凋亡蛋白Bcl-2、凋亡蛋白Bax和α7烟碱型乙酰胆碱受体（α7nAChR）的表达进行定量分析。采用蛋白质免疫印迹法（Western blot分析）评估Bcl-2、Bax、Caspase-3、IL-6、NF-κB p65、α7nAChR和磷酸化信号转导子和转录激活子3（p-STAT3）的表达。HE染色、TUNEL检测、血尿素氮（BUN）和血清肌酐（SC）检测结果表明已成功建立肾损伤模型。与对照组相比，LPS组的细胞凋亡在GTS-21处理后显著增加而后又减少。然而，脾切除术联合GTS-21治疗增加了细胞凋亡，这表明脾切除术可部分抵消GTS-21的抗凋亡作用。在用LPS处理的动物中，Cr和Cys-C的含量显著增加。GTS-21治疗后这些含量降低，但脾切除术后又升高。LPS处理后，IL-6、NF-κB p65、p-STAT3、Caspase-3和Bax的表达显著上调，而α7nAChR和Bcl-2的表达显著下调。与LPS处理的小鼠相比，脾切除术降低了IL-6、NF-κB p6物质和p-STAT3的表达，这表明脾切除术抑制了GTS-21对α7nAChR途径的激活。显然，GTS-21能有效减轻LPS诱导的肾损伤；脾切除术抑制了GTS-21的抗炎、抗凋亡活性及肾保护作用。另一方面，脾切除术减少了循环中炎性细胞因子的产生，对肾脏有一定的保护作用。因此，脾切除术对LPS诱导的AKI的影响取决于这两个方面作用的强弱。