GPR4 Knockout Improves the Neurotoxin-Induced, Caspase-Dependent Mitochondrial Apoptosis of the Dopaminergic Neuronal Cell.
GPR4 Knockout Improves the Neurotoxin-Induced, Caspase-Dependent Mitochondrial Apoptosis of the Dopaminergic Neuronal Cell.
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GPR4基因敲除改善了多巴胺能神经元细胞的神经毒素诱导的caspase依赖性线粒体凋亡。
DOI：
10.3390/ijms21207517
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发表时间：
2020-10-12
影响因子：
5.6
通讯作者：
Kim IS
中科院分区：
文献类型：
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作者：
Haque ME;Akther M;Azam S;Choi DK;Kim IS
在帕金森病中，线粒体氧化应激介导的细胞凋亡是黑质多巴胺能神经元丢失的主要原因。G蛋白偶联受体4(GPR4)，以前被认为是一个孤儿G蛋白偶联受体(GPCR4)，最近被认为是质子激活的GPCRs的成员。然而，它在神经细胞凋亡中的活性仍然不确定。在这项研究中，我们研究了GPR4在1-甲基-4-苯基吡啶离子(MPP+)和过氧化氢(H_2O_2)诱导稳定表达GPR4过表达和稳定GPR4基因敲除的人神经母细胞瘤SH-SY5Y细胞凋亡中的作用。在GPR4-OE细胞中，MPP+和H_2O_2显著上调促凋亡基因Bcl2相关X蛋白(Bax)的mRNA和蛋白表达水平，而降低抗凋亡B细胞淋巴瘤2(Bcl2)基因的表达水平。此外，MPP+还激活了caspase-3，导致聚腺苷二磷酸核糖聚合酶(PARP)的裂解，降低了线粒体膜电位(ΔΨm)。相反，过氧化氢处理显著增加GPR4-OE细胞内钙离子(Ca~(2+))和活性氧(ROS)。此外，GPR4的选择性拮抗剂NE52QQ57的化学抑制和簇状规则间隔短回文重复序列(CRISPR)/CAS9敲除GPR4降低了Bax/Bcl2比值和ROS的产生，并稳定了ΔΨm，从而保护SH-SY5Y细胞免受MPP+或H_2O_2诱导的细胞死亡。此外，GPR4基因敲除减少了磷脂酰肌醇二磷酸(PIP2)的蛋白降解，以及随后内质网(ER)储存在细胞质中的钙离子的释放。我们的结果表明，GPR4的药物抑制或基因缺失改善了神经毒素诱导的caspase依赖的线粒体凋亡途径，可能是通过调节PIP2降解介导的钙信号。因此，GPR4为帕金森病等神经退行性疾病提供了一个潜在的治疗靶点。
In Parkinson’s disease, mitochondrial oxidative stress-mediated apoptosis is a major cause of dopaminergic neuronal loss in the substantia nigra (SN). G protein-coupled receptor 4 (GPR4), previously recognised as an orphan G protein coupled-receptor (GPCR), has recently been claimed as a member of the group of proton-activated GPCRs. Its activity in neuronal apoptosis, however, remains undefined. In this study, we investigated the role of GPR4 in the 1-methyl-4-phenylpyridinium ion (MPP+) and hydrogen peroxide (H2O2)-treated apoptotic cell death of stably GPR4-overexpressing and stably GPR4-knockout human neuroblastoma SH-SY5Y cells. In GPR4-OE cells, MPP+ and H2O2 were found to significantly increase the expression levels of both mRNA and proteins of the pro-apoptotic Bcl-2-associated X protein (Bax) genes, while they decreased the anti-apoptotic B-cell lymphoma 2 (Bcl-2) genes. In addition, MPP+ treatment activated Caspase-3, leading to the cleavage of poly (ADP-ribose) polymerase (PARP) and decreasing the mitochondrial membrane potential (ΔΨm) in GPR4-OE cells. In contrast, H2O2 treatment significantly increased the intracellular calcium ions (Ca2+) and reactive oxygen species (ROS) in GPR4-OE cells. Further, chemical inhibition by NE52-QQ57, a selective antagonist of GPR4, and knockout of GPR4 by clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 decreased the Bax/Bcl-2 ratio and ROS generation, and stabilised the ΔΨm, thus protecting the SH-SY5Y cells from MPP+- or H2O2-induced apoptotic cell death. Moreover, the knockout of GPR4 decreased the proteolytic degradation of phosphatidylinositol biphosphate (PIP2) and subsequent release of the endoplasmic reticulum (ER)-stored Ca2+ in the cytosol. Our results suggest that the pharmacological inhibition or genetic deletion of GPR4 improves the neurotoxin-induced caspase-dependent mitochondrial apoptotic pathway, possibly through the modulation of PIP2 degradation-mediated calcium signalling. Therefore, GPR4 presents a potential therapeutic target for neurodegenerative disorders such as Parkinson’s disease.
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DOI：
10.1186/1478-811x-8-31
发表时间：
2010-12-22
期刊：
Cell communication and signaling : CCS
影响因子：
--
作者：
Chaitanya GV;Steven AJ;Babu PP
通讯作者：
Babu PP
缺血/再灌注诱导的 CHOP 表达促进细胞凋亡并损害肾功能恢复：酸中毒和 GPR4 的作用
影响因子：
3.7
作者：
Dong B;Zhou H;Han C;Yao J;Xu L;Zhang M;Fu Y;Xia Q
通讯作者：
Xia Q
DOI：
10.1359/jbmr.081015
发表时间：
2009-02
期刊：
Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research
影响因子：
--
作者：
Frick KK;Krieger NS;Nehrke K;Bushinsky DA
通讯作者：
Bushinsky DA
影响因子：
4.7
作者：
Hosford PS;Mosienko V;Kishi K;Jurisic G;Seuwen K;Kinzel B;Ludwig MG;Wells JA;Christie IN;Koolen L;Abdala AP;Liu BH;Gourine AV;Teschemacher AG;Kasparov S
通讯作者：
Kasparov S
影响因子：
4.6
作者：
Karthivashan G;Park SY;Kweon MH;Kim J;Haque ME;Cho DY;Kim IS;Cho EA;Ganesan P;Choi DK
通讯作者：
Choi DK