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LPS诱导小鼠中性粒细胞mGluR5的表达及mGluR5在炎症中的作用
LPS-induced expression of mGluR5 in murine neutrophils and its role in inflammation
in vitro - 第三军医大学学报 2018年40卷第22期 页码：2047-2052
- 作者机构：
400038 重庆，陆军军医大学(第三军医大学)基础医学院生物化学与分子生物学教研室
- 作者简介：
[ 何凤田，E-mail: hefengtian06@aliyun.com HE Fengtian, E-mail: hefengtian06@aliyun.com
- 基金信息：国家自然科学基金面上项目(81771693);国家自然科学基金面上项目(31370913)
DOI：10.16016/j.1000-5404.201809094
中图分类号： R364.5; R392.11; R392.3收稿：2018-09-12，
修回：2018-10-30，
纸质出版：2018-11-30
- 稿件说明：
移动端阅览
杨亭, 刘阳珷玥, 赵力, 等. LPS诱导小鼠中性粒细胞mGluR5的表达及mGluR5在炎症中的作用[J]. 第三军医大学学报, 2018,40(22):2047-2052.
Ting YANG, Yangwuyue LIU, Li ZHAO, et al. LPS-induced expression of mGluR5 in murine neutrophils and its role in inflammation
in vitro [J]. Journal of Third Military Medical University, 2018, 40(22): 2047-2052.杨亭, 刘阳珷玥, 赵力, 等. LPS诱导小鼠中性粒细胞mGluR5的表达及mGluR5在炎症中的作用[J]. 第三军医大学学报, 2018,40(22):2047-2052. DOI： 10.16016/j.1000-5404.201809094.
Ting YANG, Yangwuyue LIU, Li ZHAO, et al. LPS-induced expression of mGluR5 in murine neutrophils and its role in inflammation
in vitro [J]. Journal of Third Military Medical University, 2018, 40(22): 2047-2052. DOI： 10.16016/j.1000-5404.201809094.
摘要
目的
2
探讨脂多糖(lipopolysaccharides
LPS)诱导小鼠中性粒细胞代谢型谷氨酸受体5 (metabotropic glutamate receptor 5
mGluR5)表达的机制及其在炎症中发挥的作用。
方法
2
以原代培养mGluR5全身敲除和野生型C57BL/6J小鼠中性粒细胞为研究对象，采用ELISA及real time-PCR检测mGluR5激动剂和拮抗剂对LPS诱导的炎症反应的影响。Western blot检测LPS处理后原代小鼠中性粒细胞mGluR5蛋白及其相应下游分子表达变化情况，并用特异性拮抗剂观察能否阻断LPS诱导的生物学效应。
结果
2
特异性mGluR5的激动剂CHPG明显促进了LPS诱导的炎症反应，并且可以被mGluR5拮抗剂MPEP抑制，mGluR5基因敲除的中性粒细胞与野生型相比炎症因子水平也明显降低(
P
<
0.01)；LPS处理后的中性粒细胞mGluR5的表达明显上调，且伴随着ERK、AKT及P65的磷酸化水平明显升高(
P
<
0.05)；用特异性AKT抑制剂(LY294002)及P65抑制剂(JSH23)可以显著抑制LPS诱导mGluR5的表达及炎症反应(
P
<
0.05)
。
结论
2
LPS可通过AKT及P65通路诱导中性粒细胞mGluR5的表达，mGluR5的激活可以进一步促进炎症反应。拮抗mGluR5可以显著抑制LPS诱导的炎症反应。
Abstract
Objective
2
To investigate the mechanism of lipopolysaccharides (LPS)-induced expression of metabotropic glutamate receptor 5 (mGluR5) in murine neutrophils and explore its role in inflammation.
Methods
2
ELISA and real time-PCR were used to detect the cytokine levels in freshly isolated neutrophils derived from mGluR5 KO mice and wild type C57BL/6J mice after treatment of mGluR5 specific agonist CHPG or antagonist MPEP. The expression levels of mGluR5 and related downstream molecules were detected by Western blotting after LPS treatment. Specific downstream inhibitors were also used to verify which molecules were involved in LPS-induced biological effects.
Results
2
Activation of mGluR5 significantly promoted LPS-induced releases of cytokines
which were confirmed by ELISA and real time-PCR. The inflammatory profile(IL-1β and TNF-α) was much lower in mouse neutrophils derived from mGluR5 KO mice than those from wild type mice(
P
<
0.01). Meanwhile
Western blotting results indicated that LPS promoted the expression of mGluR5
which was accompanied with up-regulated levels of phosphorylated ERK
AKT and P65(
P
<
0.05). Specific AKT inhibitor as well as P65 inhibitor could block these LPS-induced effects(
P
<
0.05).
Conclusion
2
LPS induces mGluR5 expression in neutrophils through AKT and P65 signaling pathways. Activation of mGluR5 in neutrophils further promotes LPS-induced inflammatory responses
which might be reverted evidently by antagonizing mGluR5.
关键词
Keywords
references
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