G-protein-coupled estrogen receptor activation upregulates interleukin-1 receptor antagonist in the hippocampus after global cerebral ischemia: implications for neuronal self-defense
G-protein-coupled estrogen receptor activation upregulates interleukin-1 receptor antagonist in the hippocampus after global cerebral ischemia: implications for neuronal self-defense
复制标题
全脑缺血后 G 蛋白偶联雌激素受体激活上调海马白细胞介素 1 受体拮抗剂：对神经元自卫的影响。
DOI：
10.1186/s12974-020-1715-x
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发表时间：
2020-02-01
影响因子：
9.3
通讯作者：
Wang, Ruimin
中科院分区：
文献类型：
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作者：
Bai, Ning;Zhang, Quanguang;Wang, Ruimin
研究背景G蛋白偶联雌激素受体（GPER/GPR 30）是一种新型的膜结合雌激素受体，可在多种细胞中诱导快速的激酶信号转导。GPER的激活可以防止短暂性全脑缺血（GCI）后海马神经元的死亡，但其机制尚不清楚。在目前的研究中，我们试图解决是否GPER激活发挥有效的抗炎作用，在大鼠海马GCI后，作为一个潜在的机制，以限制神经元细胞死亡。方法采用四血管阻断法建立去势雌性SD大鼠GCI模型。采用微泵给药GPER特异性激动剂G1或拮抗剂G36，采用脑灌注药盒给药白细胞介素1 β受体拮抗剂（IL 1 RA）反义寡核苷酸（AS）。通过免疫荧光染色和Western印迹分析研究海马CA 1区IL 1 RA、NF-κ B-P65、CREB磷酸化（p-CREB）、Bcl 2、裂解的半胱天冬酶3和小胶质细胞标志物Iba 1、CD 11b以及炎性体组分NLRP 3、ASC、裂解的半胱天冬酶1和Cle-IL 1 β的蛋白表达。进行Duolink II原位邻位连接测定（PLA）以检测NLRP 3与ASC之间的相互作用。NeuN免疫荧光染色和TUNEL分析分别用于分析神经元存活和凋亡。采用巴恩斯迷宫和新物体实验比较大鼠的认知功能。结果再灌注14 d时，G1可抑制GCI诱导的海马CA 1区Iba 1和CD 11b的升高，G36可阻断G1的这种作用。G1处理还显著降低了NLRP 3-ASC-半胱天冬酶1炎性体的表达和IL 1 β活化，以及下游NF-κ B信号传导，该作用被G36施用逆转。有趣的是，G1引起了一种众所周知的内源性抗炎因子IL 1 RA的神经元的强烈升高，这被G36治疗逆转。G1还增强了海马中的p-CREB水平，这是一种已知可增强IL 1 RA表达的转录因子。最后，在体内IL 1 RA-AS废除GCI后G1的抗炎、神经保护和抗凋亡作用，并在GCI后14天逆转G1的认知增强作用。结论GPER可能通过上调IL 1 RA的表达，发挥抗炎作用，增强神经元的防御机制，从而保护GCI后的认知功能。
Background G-protein-coupled estrogen receptor (GPER/GPR30) is a novel membrane-associated estrogen receptor that can induce rapid kinase signaling in various cells. Activation of GPER can prevent hippocampal neuronal cell death following transient global cerebral ischemia (GCI), although the mechanisms remain unclear. In the current study, we sought to address whether GPER activation exerts potent anti-inflammatory effects in the rat hippocampus after GCI as a potential mechanism to limit neuronal cell death. Methods GCI was induced by four-vessel occlusion in ovariectomized female SD rats. Specific agonist G1 or antagonist G36 of GPER was administrated using minipump, and antisense oligonucleotide (AS) of interleukin-1 beta receptor antagonist (IL1RA) was administrated using brain infusion kit. Protein expression of IL1RA, NF-kappa B-P65, phosphorylation of CREB (p-CREB), Bcl2, cleaved caspase 3, and microglial markers Iba1, CD11b, as well as inflammasome components NLRP3, ASC, cleaved caspase 1, and Cle-IL1 beta in the hippocampal CA1 region were investigated by immunofluorescent staining and Western blot analysis. The Duolink II in situ proximity ligation assay (PLA) was performed to detect the interaction between NLRP3 and ASC. Immunofluorescent staining for NeuN and TUNEL analysis were used to analyze neuronal survival and apoptosis, respectively. We performed Barnes maze and Novel object tests to compare the cognitive function of the rats. Results The results showed that G1 attenuated GCI-induced elevation of Iba1 and CD11b in the hippocampal CA1 region at 14 days of reperfusion, and this effect was blocked by G36. G1 treatment also markedly decreased expression of the NLRP3-ASC-caspase 1 inflammasome and IL1 beta activation, as well as downstream NF-kappa B signaling, the effects reversed by G36 administration. Intriguingly, G1 caused a robust elevation in neurons of a well-known endogenous anti-inflammatory factor IL1RA, which was reversed by G36 treatment. G1 also enhanced p-CREB level in the hippocampus, a transcription factor known to enhance expression of IL1RA. Finally, in vivo IL1RA-AS abolished the anti-inflammatory, neuroprotective, and anti-apoptotic effects of G1 after GCI and reversed the cognitive-enhancing effects of G1 at 14 days after GCI. Conclusions Taken together, the current results suggest that GPER preserves cognitive function following GCI in part by exerting anti-inflammatory effects and enhancing the defense mechanism of neurons by upregulating IL1RA.