Upregulation of serum and glucocorticoid-regulated kinase 1 exacerbates brain injury and neurological deficits after cardiac arrest

Abstract

Cardiopulmonary arrest (CA) is the leading cause of death and disability in the United States. CA-induced brain injury is influenced by multi-factorial processes, including reduced cerebral blood flow (hypoperfusion) and neuroinflammation, which can lead to neuronal cell death and functional deficits. We have identified serum and glucocorticoid-regulated kinase-1 (SGK1) as a new target in brain ischemia previously described in the heart, liver, and kidneys (i.e. diabetes and hypertension). Our data suggest brain SGK1 mRNA and protein expression (i.e. hippocampus), presented with hypoperfusion (low cerebral blood flow) and neuroinflammation leading to further studies of the potential role of SGK1 in CA-induced brain injury. We used a 6-mins asphyxia cardiac arrest (ACA) rat model to induce global cerebral ischemia. Modulation of SGK1 was implemented via GSK650394, a SGK1 specific inhibitor (1.2 μg/kg, intracerebroventricular injection). Treatment with GSK650394 attenuated cortical hypoperfusion and neuroinflammation (via Iba1 expression) after ACA, whereas neuronal survival was enhanced in the CA1 region of the hippocampus. Learning/memory deficits were observed 3 days after ACA but ameliorated with GSK650394. SGK1 is a major contributor to ACA-induced brain injury and neurological deficits, while inhibition of SGK1 with GSK650394 provided neuroprotection against CA-induced hypoperfusion, neuroinflammation, neuronal cell death, and learning/memory deficits. Our studies could lead to a novel, therapeutic target for alleviating brain injury following cerebral ischemia.

Hepatocyte SGK1 activated by hepatic ischemia-reperfusion promotes the recurrence of liver metastasis via IL-6/STAT3

“…GSK-650394 has been validated in in vitro models to attenuate cerebral ischemia-reperfusion injury. [ 31 – 33 ] Related studies have shown that GSK-650394 exhibits antitumor activity in established breast cancer cell lines in vitro and in vivo [ 56 ]. Application of GSK-650394 to attenuates breast cancer lung metastasis in mice [ 57 ], and we wondered whether GSK-650394 had similar efficacy in CRLM.…”

Hepatocyte SGK1 activated by hepatic ischemia-reperfusion promotes the recurrence of liver metastasis via IL-6/STAT3

“…GSK-650394 has been validated in in vitro models to attenuate cerebral ischemia-reperfusion injury. [ 31 – 33 ] Related studies have shown that GSK-650394 exhibits antitumor activity in established breast cancer cell lines in vitro and in vivo [ 56 ]. Application of GSK-650394 to attenuates breast cancer lung metastasis in mice [ 57 ], and we wondered whether GSK-650394 had similar efficacy in CRLM.…”

Hippocampal differential expression underlying the neuroprotective effect of delta-9-tetrahydrocannabinol microdose on old mice

“…NeuroD2, Mc1r, Timp3 and Nfkbia were all upregulated at 5 weeks after treatment, and were associated with protection against neuroinflammation, promotion of cell survival, and attenuation of neuronal and cognitive damage in numerous animal models ( Fridmacher et al, 2003 ; Gibb et al, 2015 ; Fu et al, 2020 ; Pang and Shi, 2021 ; Xu et al, 2021 ). Among these upregulated genes, Sgk1 in particular was associated with cell survival in the aging brain ( Sahin et al, 2013 ), and highly protective against a broad range of neurotoxins ( Iqbal et al, 2015 ) and pathologies ( Lee et al, 2020 ; Martin-Batista et al, 2021 ).…”

The role of serum/glucocorticoid-regulated kinase 1 in brain function following cerebral ischemia

“…The number of animals per group was determined based on our previous publications 6,19 –22,26,38 –40 and power analysis with p < 0.05 (a power of 0.95) (homepage.univie.ac.at/robin.rist/samplesize.php?test=anova). We also adjusted animal numbers based on standard deviation if standard deviation was higher than projected to ensure sufficient power.…”

“…Per our recent publications, 5 animals per group is sufficient for studies related to cerebral blood flow measurements. 19,20,26,38,39 Animal behavior studies required at least 6 animals per group to achieve statistical power, 6,21,39 while a minimum sample size of 3–4 is required to achieve the desired level of confidence for brain histology, inflammatory arrays, and Seahorse analyzer experiment. 21,22,26,40 As for the Ndrg1/SOX10 study (Figure 5), we compared the overall differences (across 1, 3, and 7 days after cardiac arrest) between cardiac arrest and treatment groups (via two-way ANOVA with Tukey's post-hoc).…”

“…Specific SGK1 inhibitor (GSK650394, Tocris, 3572) was dissolved in dimethyl sulfoxide and diluted from a stock solution (1 mM) with sterile water to the desired concentration (1.2 lg/kg). 6 GSK650394 was administered 5 minutes before CA(rat) surgery via intracerebroventricular injection to the third ventricle (2.8 mm posterior to bregma, 0 mm lateral to midline, and 8.3 mm deep). For post-treatment experiments, GSK650394 was delivered directly into the cerebrospinal fluid via intrathecal injection (between the groove of L5 and L6 vertebrae) 30 mins, 1, and 3 days after CA surgery via a 25 ml Hamilton syringe with a 30G 0.5-inch needle.…”

Hepatocyte SGK1 activated by hepatic ischemia-reperfusion promotes the recurrence of liver metastasis via IL-6/STAT3

“…GSK-650394 has been validated in in vitro models to attenuate cerebral ischemia-reperfusion injury. [ 31 – 33 ] Related studies have shown that GSK-650394 exhibits antitumor activity in established breast cancer cell lines in vitro and in vivo [ 56 ]. Application of GSK-650394 to attenuates breast cancer lung metastasis in mice [ 57 ], and we wondered whether GSK-650394 had similar efficacy in CRLM.…”

Hippocampal differential expression underlying the neuroprotective effect of delta-9-tetrahydrocannabinol microdose on old mice

“…NeuroD2, Mc1r, Timp3 and Nfkbia were all upregulated at 5 weeks after treatment, and were associated with protection against neuroinflammation, promotion of cell survival, and attenuation of neuronal and cognitive damage in numerous animal models ( Fridmacher et al, 2003 ; Gibb et al, 2015 ; Fu et al, 2020 ; Pang and Shi, 2021 ; Xu et al, 2021 ). Among these upregulated genes, Sgk1 in particular was associated with cell survival in the aging brain ( Sahin et al, 2013 ), and highly protective against a broad range of neurotoxins ( Iqbal et al, 2015 ) and pathologies ( Lee et al, 2020 ; Martin-Batista et al, 2021 ).…”

The role of serum/glucocorticoid-regulated kinase 1 in brain function following cerebral ischemia

“…The number of animals per group was determined based on our previous publications 6,19 –22,26,38 –40 and power analysis with p < 0.05 (a power of 0.95) (homepage.univie.ac.at/robin.rist/samplesize.php?test=anova). We also adjusted animal numbers based on standard deviation if standard deviation was higher than projected to ensure sufficient power.…”

“…Per our recent publications, 5 animals per group is sufficient for studies related to cerebral blood flow measurements. 19,20,26,38,39 Animal behavior studies required at least 6 animals per group to achieve statistical power, 6,21,39 while a minimum sample size of 3–4 is required to achieve the desired level of confidence for brain histology, inflammatory arrays, and Seahorse analyzer experiment. 21,22,26,40 As for the Ndrg1/SOX10 study (Figure 5), we compared the overall differences (across 1, 3, and 7 days after cardiac arrest) between cardiac arrest and treatment groups (via two-way ANOVA with Tukey's post-hoc).…”

“…Specific SGK1 inhibitor (GSK650394, Tocris, 3572) was dissolved in dimethyl sulfoxide and diluted from a stock solution (1 mM) with sterile water to the desired concentration (1.2 lg/kg). 6 GSK650394 was administered 5 minutes before CA(rat) surgery via intracerebroventricular injection to the third ventricle (2.8 mm posterior to bregma, 0 mm lateral to midline, and 8.3 mm deep). For post-treatment experiments, GSK650394 was delivered directly into the cerebrospinal fluid via intrathecal injection (between the groove of L5 and L6 vertebrae) 30 mins, 1, and 3 days after CA surgery via a 25 ml Hamilton syringe with a 30G 0.5-inch needle.…”

Hepatocyte SGK1 activated by hepatic ischemia-reperfusion promotes the recurrence of liver metastasis via IL-6/STAT3

“…GSK-650394 has been validated in in vitro models to attenuate cerebral ischemia-reperfusion injury. [ 31 – 33 ] Related studies have shown that GSK-650394 exhibits antitumor activity in established breast cancer cell lines in vitro and in vivo [ 56 ]. Application of GSK-650394 to attenuates breast cancer lung metastasis in mice [ 57 ], and we wondered whether GSK-650394 had similar efficacy in CRLM.…”

Hippocampal differential expression underlying the neuroprotective effect of delta-9-tetrahydrocannabinol microdose on old mice

“…NeuroD2, Mc1r, Timp3 and Nfkbia were all upregulated at 5 weeks after treatment, and were associated with protection against neuroinflammation, promotion of cell survival, and attenuation of neuronal and cognitive damage in numerous animal models ( Fridmacher et al, 2003 ; Gibb et al, 2015 ; Fu et al, 2020 ; Pang and Shi, 2021 ; Xu et al, 2021 ). Among these upregulated genes, Sgk1 in particular was associated with cell survival in the aging brain ( Sahin et al, 2013 ), and highly protective against a broad range of neurotoxins ( Iqbal et al, 2015 ) and pathologies ( Lee et al, 2020 ; Martin-Batista et al, 2021 ).…”

The role of serum/glucocorticoid-regulated kinase 1 in brain function following cerebral ischemia

“…The number of animals per group was determined based on our previous publications 6,19 –22,26,38 –40 and power analysis with p < 0.05 (a power of 0.95) (homepage.univie.ac.at/robin.rist/samplesize.php?test=anova). We also adjusted animal numbers based on standard deviation if standard deviation was higher than projected to ensure sufficient power.…”

“…Per our recent publications, 5 animals per group is sufficient for studies related to cerebral blood flow measurements. 19,20,26,38,39 Animal behavior studies required at least 6 animals per group to achieve statistical power, 6,21,39 while a minimum sample size of 3–4 is required to achieve the desired level of confidence for brain histology, inflammatory arrays, and Seahorse analyzer experiment. 21,22,26,40 As for the Ndrg1/SOX10 study (Figure 5), we compared the overall differences (across 1, 3, and 7 days after cardiac arrest) between cardiac arrest and treatment groups (via two-way ANOVA with Tukey's post-hoc).…”

“…Specific SGK1 inhibitor (GSK650394, Tocris, 3572) was dissolved in dimethyl sulfoxide and diluted from a stock solution (1 mM) with sterile water to the desired concentration (1.2 lg/kg). 6 GSK650394 was administered 5 minutes before CA(rat) surgery via intracerebroventricular injection to the third ventricle (2.8 mm posterior to bregma, 0 mm lateral to midline, and 8.3 mm deep). For post-treatment experiments, GSK650394 was delivered directly into the cerebrospinal fluid via intrathecal injection (between the groove of L5 and L6 vertebrae) 30 mins, 1, and 3 days after CA surgery via a 25 ml Hamilton syringe with a 30G 0.5-inch needle.…”