Objective:To investigate the effects of glimepiride on cerebral edema and apoptosis in a rat model of sub-arachnoid hemorrhage(SAH).Methods:An SAH model was established in rats using the internal carotid artery punc-ture method.Low-dose glimepiride was administered via intraperitoneal injection.Neurological deficits were assessed u-sing the modified Neurological Severity Score(mNSS),and brain water content was evaluated by measuring the wet-dry weight ratio of brain tissue.Cortical tissue from the surgical side was collected to measure the mRNA expression of sul-fonylurea receptor 1(SUR1)and transient receptor potential melastatin 4(TRPM4)using RT-qPCR,while the protein expression of Bcl-2 and Bax was detected by Western blot.Results:SAH rats exhibited impaired neurological function,increased brain water content,upregulation of SUR1 and TRPM4 mRNA expression,and a decreased Bcl-2/Bax ratio.Low-dose glimepiride did not affect blood glucose levels but significantly improved neurological function and reduced cerebral edema in SAH rats.Although glimepiride had no significant effect on SUR1 and TRPM4 mRNA expression,it increased the Bcl-2/Bax ratio.Conclusion:Glimepiride alleviates cerebral edema and inhibits apoptosis in SAH model rats by suppressing the SUR1-TRPM4 channel.

To investigate the impact of miR-142-5p on intestinal epithelial cells,an expression vector for overexpressing miR-142-5p lentivirus was constructed and administered to mice to observe pathological changes in their colon tissue.The miR-142-5p gene was integrated into the green fluo-rescent lentiviral vector plenti-CMV/TO eGFP-Puro(plenti),and the resulting recombinant vec-tor was named plenti miR-142-5p following confirmation through sequencing.293T cells were transfected with the recombinant plasmid and packaging-assisted plasmid,yielding a packaged miR-142-5p overexpressing lentiviral vector(LV-miR-142-5p).The recombinant lentivirus suspen-sion was collected and concentrated via ultrafast centrifugal precipitation.Virus titer was deter-mined using real-time PCR.The virus suspension was then injected into the tail vein of 6-8-week-old BALB/c mice,after which their colonic tissues were dissected for HE staining and microscopic observation.The results showed that miR-142-5p homologous recombination into plenti vector and sequencing results were consistent with the expected sequence.plenti-miR-142-5p was co-transfect-ed with the packaging helper plasmid in 293T cells,and the successful transfection was confirmed based on the green fluorescence expression,and the real-time PCR results showed that the lentiviral titer was 1.23 × 109 IU/mL.Colon tissue slices from infected mice exhibited compromised colon integrity and significant inflammatory response.It was proved that LV-miR-142-5p was suc-cessfully constructed,and miR-142-5p caused damage to intestinal epithelial cells of mice.

Objective:To explore the effect of melatonin on mitophagy in the brain tissue of rats with early subarach-noid hemorrhage(SAH).Methods:The rat model of SAH was established by internal carotid artery puncture and trea-ted with melatonin by intraperitoneal injection.At 24 hours after the operation,the neurological function of rats was de-tected by mNSS score and balance beam test.The contents of reactive oxygen species(ROS),malondialdehyde(MDA),and glutathione(GSH)and in brain tissue were detected by commercial reagent kits.The expression changes of PINK1/Parkin,microtubule-associated protein1A/1B-light chain3(LC3)and p62 in brain tissue were detected by Western blot.Results:Melatonin can significantly improve the neurological function of SAH rats,reduce the con-tents of MDA(P<0.05)and ROS(P<0.05),and increase the content of GSH(P<0.05).Meanwhile,Western blot detection indicated that melatonin could increase the expression of PINK1 and Parkin(P<0.05),increase the ratio of LC3-II/LC3-I(P<0.05),and reduce the expression of p62(P<0.05).Conclusion:Melatonin alleviates early brain injury in rats with subarachnoid hemorrhage through PINK1/Parkin-mediated mitophagy.

Objective:To evaluate the effect of deep brain stimulation(DBS)of the lateral cerebellar nucleus(LCN)on synaptic plasticity in a traumatic brain injury(TBI)rat model of the medial prefrontal cortex(mPFC).Methods:A controllable mPFC injury rat model was prepared using a hydraulic impact method.Two weeks after inju-ry,stimulation electrodes were implanted into the left LCN.Electrical stimulation was initiated at the 4th week after injury and continued for 30 days.The learning and memory function of the rats was evaluated using the Morris water maze.RT-qPCR and Western blot were used to detect the mRNA and protein expressions of brain-derived neurotrophic factor(BDNF),postsynaptic density protein 95(PSD95),and growth-associated protein 43(GAP43)in the mPFC region.Results:The learning and memory function of the mPFC-injured rats significantly decreased.The expressions of BDNF,PSD95,and GAP43 in mPFC decreased.DBS treatment significantly enhanced the learning and memory ability of mPFC-injured rats,and simultaneously upregulated the expressions of BDNF,PSD95,and GAP43 in mPFC.Conclusion:LCN DBS is an effective method for treating cognitive impairment caused by TBI,and it may achieve this through activating the neural plasticity mechanism of the mPFC.

Objective:To evaluate the effect of deep brain stimulation(DBS)of the lateral cerebellar nucleus(LCN)on synaptic plasticity in a traumatic brain injury(TBI)rat model of the medial prefrontal cortex(mPFC).Methods:A controllable mPFC injury rat model was prepared using a hydraulic impact method.Two weeks after inju-ry,stimulation electrodes were implanted into the left LCN.Electrical stimulation was initiated at the 4th week after injury and continued for 30 days.The learning and memory function of the rats was evaluated using the Morris water maze.RT-qPCR and Western blot were used to detect the mRNA and protein expressions of brain-derived neurotrophic factor(BDNF),postsynaptic density protein 95(PSD95),and growth-associated protein 43(GAP43)in the mPFC region.Results:The learning and memory function of the mPFC-injured rats significantly decreased.The expressions of BDNF,PSD95,and GAP43 in mPFC decreased.DBS treatment significantly enhanced the learning and memory ability of mPFC-injured rats,and simultaneously upregulated the expressions of BDNF,PSD95,and GAP43 in mPFC.Conclusion:LCN DBS is an effective method for treating cognitive impairment caused by TBI,and it may achieve this through activating the neural plasticity mechanism of the mPFC.

Objective:To investigate the effects of glimepiride on cerebral edema and apoptosis in a rat model of sub-arachnoid hemorrhage(SAH).Methods:An SAH model was established in rats using the internal carotid artery punc-ture method.Low-dose glimepiride was administered via intraperitoneal injection.Neurological deficits were assessed u-sing the modified Neurological Severity Score(mNSS),and brain water content was evaluated by measuring the wet-dry weight ratio of brain tissue.Cortical tissue from the surgical side was collected to measure the mRNA expression of sul-fonylurea receptor 1(SUR1)and transient receptor potential melastatin 4(TRPM4)using RT-qPCR,while the protein expression of Bcl-2 and Bax was detected by Western blot.Results:SAH rats exhibited impaired neurological function,increased brain water content,upregulation of SUR1 and TRPM4 mRNA expression,and a decreased Bcl-2/Bax ratio.Low-dose glimepiride did not affect blood glucose levels but significantly improved neurological function and reduced cerebral edema in SAH rats.Although glimepiride had no significant effect on SUR1 and TRPM4 mRNA expression,it increased the Bcl-2/Bax ratio.Conclusion:Glimepiride alleviates cerebral edema and inhibits apoptosis in SAH model rats by suppressing the SUR1-TRPM4 channel.

To investigate the impact of miR-142-5p on intestinal epithelial cells,an expression vector for overexpressing miR-142-5p lentivirus was constructed and administered to mice to observe pathological changes in their colon tissue.The miR-142-5p gene was integrated into the green fluo-rescent lentiviral vector plenti-CMV/TO eGFP-Puro(plenti),and the resulting recombinant vec-tor was named plenti miR-142-5p following confirmation through sequencing.293T cells were transfected with the recombinant plasmid and packaging-assisted plasmid,yielding a packaged miR-142-5p overexpressing lentiviral vector(LV-miR-142-5p).The recombinant lentivirus suspen-sion was collected and concentrated via ultrafast centrifugal precipitation.Virus titer was deter-mined using real-time PCR.The virus suspension was then injected into the tail vein of 6-8-week-old BALB/c mice,after which their colonic tissues were dissected for HE staining and microscopic observation.The results showed that miR-142-5p homologous recombination into plenti vector and sequencing results were consistent with the expected sequence.plenti-miR-142-5p was co-transfect-ed with the packaging helper plasmid in 293T cells,and the successful transfection was confirmed based on the green fluorescence expression,and the real-time PCR results showed that the lentiviral titer was 1.23 × 109 IU/mL.Colon tissue slices from infected mice exhibited compromised colon integrity and significant inflammatory response.It was proved that LV-miR-142-5p was suc-cessfully constructed,and miR-142-5p caused damage to intestinal epithelial cells of mice.

Objective:To explore the effect of melatonin on mitophagy in the brain tissue of rats with early subarach-noid hemorrhage(SAH).Methods:The rat model of SAH was established by internal carotid artery puncture and trea-ted with melatonin by intraperitoneal injection.At 24 hours after the operation,the neurological function of rats was de-tected by mNSS score and balance beam test.The contents of reactive oxygen species(ROS),malondialdehyde(MDA),and glutathione(GSH)and in brain tissue were detected by commercial reagent kits.The expression changes of PINK1/Parkin,microtubule-associated protein1A/1B-light chain3(LC3)and p62 in brain tissue were detected by Western blot.Results:Melatonin can significantly improve the neurological function of SAH rats,reduce the con-tents of MDA(P<0.05)and ROS(P<0.05),and increase the content of GSH(P<0.05).Meanwhile,Western blot detection indicated that melatonin could increase the expression of PINK1 and Parkin(P<0.05),increase the ratio of LC3-II/LC3-I(P<0.05),and reduce the expression of p62(P<0.05).Conclusion:Melatonin alleviates early brain injury in rats with subarachnoid hemorrhage through PINK1/Parkin-mediated mitophagy.

Objective:To investigate the effect of minocycline on neuroinflammation of rats with post-traumatic stress disorder(PTSD).Methods:The rat model of PTSD was prepared by a single prolonged stress(SPS)method,and the rats were treated with minocycline(PTSD+Mino group)or normal saline(PTSD group)by gavage.The behavioral changes of rats were detected by light-dark box test.The expression of ionized calcium-binding adapter molecule 1(Iba-1)in hippocampus was detected by immunohistochemical staining.The contents of IL-1β and TNF-α in hippocampus were detected by ELISA,and the expression levels of IL-1β and TNF-α mRNA in hippocampus were detected by real-time RT-PCR(qRT-PCR).Results:After 3 days of SPS stimulation,the anxiety-like behavior of rats was obvious,the expression of Iba-1 in hippocampus was increased,and the contents of IL-1β and TNF-α in hippocampus were in-creased.Minocycline treatment significantly reduced anxiety-like behavior and decreased the expression of Iba-1 in the hippocampus of PTSD rats.Meanwhile,minocycline treatment also decreased the levels of IL-1β and TNF-α mRNA and protein in the hippocampus.Conclusion:Minocycline can improve the anxiety-like behavior of PTSD rats by inhibiting the activation of microglia.

Objective:To study the role of DDX3X/NF-κB pathway in early neuronal apoptosis in subarachnoid hemorrhage(SAH)mice.Methods:The mouse model of SAH was established by internal carotid artery puncture,and the neurological function score of the mice was evaluated.The DDX3X expression was knocked down using recombinant lentivirus expressing DDX3X targeted shRNA(Lv-shDDX3X),or the NF-κB pathway was inhibited by NF-κB-IN-1(IN-1).Western Blot was used to detect the expression of DDX3X and NF-κB(p65)in mouse cortex.TUNEL/NeuN staining was used to detect the apoptosis of cerebral cortex neurons.Results:Twenty-four hours after SAH operation,the neurological function of mice was significantly impaired(P＜0.05).While the expression of DDX3X was signifi-cantly increased and the expression of NF-κB(p65)was significantly decreased in the cortex(P＜0.05).When the DDX3X expression is knocked down firstly,then SAH surgery is performed.The neurological function of mice was sig-nificantly recovered,and the expression of NF-κB(p65)protein was significantly higher than that in SAH group(P＜0.05);If the NF-κB activity was inhibited by IN-1 while DDX3X knockdown,there is no significant recovery of neuro-logical function in SAH mice.TUNEL/NeuN staining showed that the number of TUNEL-positive neurons in the brain tissue after DDX3X knockdown was less than that in the SAH group(P＜0.05),while the number of TUNEL-positive neurons was not significantly reduced when IN-1 was used to inhibit NF-κB activity at the same time of DDX3X knock-down.Conclusion:DDX3X/NF-κB mediated cell death in mice with early brain injury after SAH.