VIP对LPS应激的小鼠成纤维细胞TREM-2表达的影响及其机制

李淑芬; 宋卓慧; 杨慧慧; 刘永平; 熊建兵; 管茶香; 孙国瑛

doi:10.13418/j.issn.1001-165x.2018.04.010

中国临床解剖学杂志 ›› 2018, Vol. 36 ›› Issue (4) : 402-407. DOI: 10.13418/j.issn.1001-165x.2018.04.010

实验研究

VIP对LPS应激的小鼠成纤维细胞TREM-2表达的影响及其机制

李淑芬¹，　宋卓慧¹，　杨慧慧²，　刘永平²，　熊建兵²，　管茶香²，　孙国瑛^{2, 3}

作者信息 +

Effect of VIP on the expression of TREM-2 in mouse fibroblasts with LPS stress and its mechanism

LI Shu-fen¹, SONG Zhuo-hui¹, YANG Hui-hui², LIU Yong-ping², XIONG Jian-bing², GUAN Cha-xiang², SUN Guo-ying ^{2, 3}

Author information +

文章历史 +

摘要

目的　观察血管活性肠肽（vasoactive intestinal peptide，VIP）对脂多糖（lipopolysaccharides, LPS）应激的小鼠成纤维细胞髓样细胞表达触发受体-2（TREM-2）表达的影响，并初步探讨其信号转导通路。方法　利用LPS腹腔注射建立急性肺损伤（ALI）小鼠模型；采用VIP慢病毒气管滴注，qPCR检测肺组织TREM-2的表达。选用qPCR和流式细胞术检测VIP对LPS应激的小鼠成纤维细胞TREM- 2表达的影响；并观察PKC信号通路阻断剂（H-7）、PKA信号通路阻断剂（H- 89）、MAPK信号通路阻断剂（PD98059）和CaM信号通路阻断剂（W-7）对VIP调控TREM- 2表达的影响。结果　ALI时小鼠肺组织TREM-2 mRNA表达降低，而VIP可上调肺组织TREM- 2 mRNA的表达。LPS下调小鼠成纤维细胞TREM- 2 mRNA的表达，VIP可呈时间依赖性上调TREM- 2 mRNA的表达（0、3 、6 、12和24 h），且在6 h达到峰值；并呈剂量相关性上调TREM- 2 mRNA的表达（10-10、10-9、10-8和10-7 mol/L），以10-8 mol/L作用最明显。VIP对LPS应激6 h增加小鼠成纤维细胞TREM-2 mRNA和蛋白表达的效应可被H-7、PD98059以及W- 7所阻断。结论　LPS下调小鼠成纤维细胞TREM-2的表达，而VIP可上调LPS应激的小鼠成纤维细胞TREM- 2 mRNA的表达，其胞内信号转导途径可能与PKC、MAPK及CaM有关。

Abstract

Objective　To observe the effect of VIP(vasoactive intestinal peptide) on TREM-2 expression in mouse fibroblasts induced by LPS(lipopolysaccharides) and to explore its possible signaling pathway. Methods　ALI animal model was established by intraperitoneal injection of LPS, and VIP lentivirus tracheal infusion was used, and the expression of TREM-2 in lung tissue was detected by qPCR. qPCR and flow cytometry were used to detect the effect of VIP on TREM-2 expression in LPS-induced fibroblasts, and to observe the effects of PKC signal pathway blocker (H-7), PKA signal pathway blocker (H-89), MAPK signal pathway blocker (PD98059) and CaM signal pathway blocker (W-7) on the regulation of VIP expression TREM-2.　Results　The TREM-2 mRNA expression of lung tissue in mice decreased in ALI, while VIP could increase TREM-2 mRNA expression in lung tissue. LPS down-regulated the expression of TREM-2 mRNA in mice fibroblast, and VIP up-regulated the expression of TREM-2 mRNA in a time-dependent manner (0 h, 3 h, 6 h, 12 h, 24 h), and reached the peak at 6 h. VIP dose related with the TREM-2 mRNA expression (10-10 mol/L, 10-9 mol/L, 10-8 mol/L, 10-7 mol/L). The up-regulation effect of VIP on TREM-2 mRNA and protein expression in mouse fibroblasts (stress by LPS for 6 h) could be blocked by H-7, PD98059, and W-7. 　Conclusion　LPS can down-regulate the TREM-2 expression of mouse fibroblasts, while VIP can up-regulate the expression of TREM-2 in LPS induced fibroblasts, and the intracellular signal transduction pathway may be related to PKC, MAPK and CaM.

导出引用

李淑芬,宋卓慧,杨慧慧,刘永平,熊建兵,管茶香,孙国瑛. VIP对LPS应激的小鼠成纤维细胞TREM-2表达的影响及其机制[J]. 中国临床解剖学杂志. 2018, 36(4): 402-407 https://doi.org/10.13418/j.issn.1001-165x.2018.04.010

LI Shu-fen, SONG Zhuo-hui, YANG Hui-hui, LIU Yong-ping, XIONG Jian-bing, GUAN Cha-xiang, SUN Guo-ying. Effect of VIP on the expression of TREM-2 in mouse fibroblasts with LPS stress and its mechanism[J]. Chinese Journal of Clinical Anatomy. 2018, 36(4): 402-407 https://doi.org/10.13418/j.issn.1001-165x.2018.04.010

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