Interactions between brain-resident and peripheral infiltrated immune cells are thought to contribute to neuroplasticity after cerebral ischemia. However, conventional bulk sequencing makes it challenging to depict this complex immune network. Using single-cell RNA sequencing, we mapped compositional and transcriptional features of peri-infarct immune cells. Microglia were the predominant cell type in the peri-infarct region, displaying a more diverse activation pattern than the typical pro- and anti-inflammatory state, with axon tract-associated microglia (ATMs) being associated with neuronal regeneration. Trajectory inference suggested that infiltrated monocyte-derived macrophages (MDMs) exhibited a gradual fate trajectory transition to activated MDMs. Inter-cellular crosstalk between MDMs and microglia orchestrated anti-inflammatory and repair-promoting microglia phenotypes and promoted post-stroke neurogenesis, with SOX2 and related Akt/CREB signaling as the underlying mechanisms. This description of the brain's immune landscape and its relationship with neurogenesis provides new insight into promoting neural repair by regulating neuroinflammatory responses.
Humans ; Ischemic Stroke ; Brain/metabolism* ; Macrophages ; Brain Ischemia/metabolism* ; Microglia/metabolism* ; Gene Expression Profiling ; Anti-Inflammatory Agents ; Neuronal Plasticity/physiology* ; Infarction/metabolism*

Objective:To investigate effects and underlying mechanisms of glutathione persulfate (GSSH) on the level of testosterone in male obese mice.Methods:Totally 45 mice were divided into 3 groups on average. Low-fat diet (LFD)+normal saline (NS) group: 15 mice were fed with LFD for 10 weeks, followed by LFD together with daily intraperitoneal injection of saline for 45 d; high-fat diet (HFD)+NS group: 15 mice were fed with high-fat diet for 10 weeks, followed by HFD and daily intraperitoneal injection of NS for 45 d; HFD+GSSH group: 15 mice were fed with HFD for 10 weeks, followed by a HFD for 45 d and daily intraperitoneal injection of GSSH (200 mg/kg). After the treatment, all mice were killed with their necks-severed, testis and serum were taken out from the mice. Serum levels of testosterone and malondialdehyde (MDA), the mRNA levels of key enzymes for testosterone synthesis ( StAR, 3β- HSD, Cyp11a1 and Cyp17a1) were measured by RT-PCR. The testicular protein levels of StAR, 3β-HSD, NR5A1 and EHD3 were measured by Western blotting assay. Protein levels of NR5A1, SOD and Nrf2 were measured in mouse Leydig TM-3 cells that were treated with 50 μmol/L and 100 μmol/L GSSH, respectively, following with treatment with 100 μmol/L H 2O 2 . Results:1) After treatment, the body weight of mice in HFD+GSSH group did not change significantly, while the body weight of mice in HFD+NS group raised by 24.53% (from 32.46 g to 40.43 g) during the 45-day-intraperitoneal injection ( P=0.002). 2) Serum level of testosterone in HFD+NS group [(12.9±1.7) μg/L] was significantly lower than that in LFD+NS group [(18.3±1.2) μg/L, P=0.020]. However, serum level of testosterone in HFD+GSSH group was (25.42±2.1) μg/L, which was significantly higher than that in HFD+NS group ( P=0.030). The RT-PCR test results showed that compared with LFD+NS group, the expression levels of all key genes involved in testosterone synthesis ( StAR, 3β- HSD, Cyp11a1, Cyp17a1) showed a significant decrease in HFD+NS group ( P=0.003, P=0.007, P<0.001, P<0.001). The expression levels of these genes were restored in the mouse testes of HFD+GSSH group ( P=0.002, P<0.001, P<0.001, P=0.006). 3) Similarly, compared with LFD+NS group [(9.00±1.59) nmol/mL], the serum MDA level of HFD+NS group [(10.61±1.73) nmol/mL] raised significantly ( P=0.016), while GSSH reversed the raised HFD+NS high level of serum MDA in HFD+GSSH group [(9.23±0.94) nmol/mL, P=0.048]. 4) Both levels of NR5A1, EHD3, StAR, and 3β-HSD were reduced in HFD+NS group ( P=0.002, P=0.012, P=0.004, P=0.043), but their levels were significantly restored in HFD+GSSH group ( P<0.001, P=0.017, P=0.004, P<0.001). 5) The levels of NR5A1, Nrf2 and SOD were obviously down-regulated in TM3 cells treated with H 2O 2 ( P<0.001, P=0.002, P=0.004). Conclusion:GSSH can raise serum level of testosterone in HFD-fed mice by up-regulating expression of genes which are important for testicular testosterone biosynthesis.

Objective:To investigate effects and underlying mechanisms of glutathione persulfate (GSSH) on the level of testosterone in male obese mice.Methods:Totally 45 mice were divided into 3 groups on average. Low-fat diet (LFD)+normal saline (NS) group: 15 mice were fed with LFD for 10 weeks, followed by LFD together with daily intraperitoneal injection of saline for 45 d; high-fat diet (HFD)+NS group: 15 mice were fed with high-fat diet for 10 weeks, followed by HFD and daily intraperitoneal injection of NS for 45 d; HFD+GSSH group: 15 mice were fed with HFD for 10 weeks, followed by a HFD for 45 d and daily intraperitoneal injection of GSSH (200 mg/kg). After the treatment, all mice were killed with their necks-severed, testis and serum were taken out from the mice. Serum levels of testosterone and malondialdehyde (MDA), the mRNA levels of key enzymes for testosterone synthesis ( StAR, 3β- HSD, Cyp11a1 and Cyp17a1) were measured by RT-PCR. The testicular protein levels of StAR, 3β-HSD, NR5A1 and EHD3 were measured by Western blotting assay. Protein levels of NR5A1, SOD and Nrf2 were measured in mouse Leydig TM-3 cells that were treated with 50 μmol/L and 100 μmol/L GSSH, respectively, following with treatment with 100 μmol/L H 2O 2 . Results:1) After treatment, the body weight of mice in HFD+GSSH group did not change significantly, while the body weight of mice in HFD+NS group raised by 24.53% (from 32.46 g to 40.43 g) during the 45-day-intraperitoneal injection ( P=0.002). 2) Serum level of testosterone in HFD+NS group [(12.9±1.7) μg/L] was significantly lower than that in LFD+NS group [(18.3±1.2) μg/L, P=0.020]. However, serum level of testosterone in HFD+GSSH group was (25.42±2.1) μg/L, which was significantly higher than that in HFD+NS group ( P=0.030). The RT-PCR test results showed that compared with LFD+NS group, the expression levels of all key genes involved in testosterone synthesis ( StAR, 3β- HSD, Cyp11a1, Cyp17a1) showed a significant decrease in HFD+NS group ( P=0.003, P=0.007, P<0.001, P<0.001). The expression levels of these genes were restored in the mouse testes of HFD+GSSH group ( P=0.002, P<0.001, P<0.001, P=0.006). 3) Similarly, compared with LFD+NS group [(9.00±1.59) nmol/mL], the serum MDA level of HFD+NS group [(10.61±1.73) nmol/mL] raised significantly ( P=0.016), while GSSH reversed the raised HFD+NS high level of serum MDA in HFD+GSSH group [(9.23±0.94) nmol/mL, P=0.048]. 4) Both levels of NR5A1, EHD3, StAR, and 3β-HSD were reduced in HFD+NS group ( P=0.002, P=0.012, P=0.004, P=0.043), but their levels were significantly restored in HFD+GSSH group ( P<0.001, P=0.017, P=0.004, P<0.001). 5) The levels of NR5A1, Nrf2 and SOD were obviously down-regulated in TM3 cells treated with H 2O 2 ( P<0.001, P=0.002, P=0.004). Conclusion:GSSH can raise serum level of testosterone in HFD-fed mice by up-regulating expression of genes which are important for testicular testosterone biosynthesis.