Objective:To explore the related mechanism of miR-34a affecting epilepsy.Methods:MicroRNA microarray chip and RT-PCR were used to analyze the expression of miR-34a in epileptic rats,and observe the relationship betwee...Objective:To explore the related mechanism of miR-34a affecting epilepsy.Methods:MicroRNA microarray chip and RT-PCR were used to analyze the expression of miR-34a in epileptic rats,and observe the relationship between miR-34a and seizure frequency and EEG.Predicted the target gene of miR-34a,used The dual-luciferase reporter gene assay was used to observe its binding to the target gene of miR-34a,the effect of overexpression and inhibition of miR-34a on the target gene,the effects of overexpression and inhibition of target genes on downstream factors.Results:Microarray chip and RT-PCR detection showed that miR-34a was highly expressed in epilepsy(P<0.05),and miR-34a was positively correlated with seizure frequency(R=0.783,P=0.003),and inhibition of miR-34a inhibited the seizure frequency of epilepsy(P<0.05).The online databases TargetScan and miRDB show that miR-34a and BCL-2 have a predicted binding region,and the dual-luciferase reporter gene experiment proves that the two have a binding relationship.When miR-34a was overexpressed,BCL-2 expression was down-regulated,and conversely,when miR-34a was inhibited,BCL-2 expression was up-regulated.NLRP1 and downstream related factors Caspase-3,Caspase-1,IL-1βand IL-18 were highly expressed in the brain tissue of epileptic rats(P<0.05).When BCL-2 was inhibited,the expression levels of NLRP1 and downstream related factors were increased,and when BCL-2 was inhibited,the result was the opposite.Conclusion:The highly expressed miR-34a promotes epilepsy by inhibiting BCL-2 and activating the NLRP1 inflammasome.展开更多
We studied the effects of Ganoderma lucidum spore powder on Bax and Bcl-2 expression and neuronal apoptosis in pentylenetetrazole-kindled epileptic rats. Sixty adult rats were randomly divided into a control group, an...We studied the effects of Ganoderma lucidum spore powder on Bax and Bcl-2 expression and neuronal apoptosis in pentylenetetrazole-kindled epileptic rats. Sixty adult rats were randomly divided into a control group, an epileptic group （kindled） and three medication groups （150,300 450 mg/kg given to kindled rats）. Bax and Bcl-2 immunohistochemistry and TUNEL labeling showed that the number of Bax- and TUNEL-positive cells in the hippocampus and cerebral cortex decreased significantly in the high-dose medication group, while the number of Bcl-2 immunoreactive cells increased. The Morris water maze test showed that high-dose treatment significantly shortened escape latency and increased spatial probe trial performance. Our findings indicate that a high dose of Ganoderma lucidum spore powder upregulates the expression of antiapoptotic Bcl-2 protein in the hippocampus and cerebral cortex, inhibits proapoptotic Bax expression, and decreases seizure-induced neuronal apoptosis. Further, Ganoderma lucidum appears to protect against epilepsy-related learning and memory impairments.展开更多
Objective: To investigate the expression of miRNA21 in hippocampus of rats with pentylenetetrazol-induced epilepsy and its possible mechanism. Methods: Twenty-four healthy male wister rats was randomly divided into tw...Objective: To investigate the expression of miRNA21 in hippocampus of rats with pentylenetetrazol-induced epilepsy and its possible mechanism. Methods: Twenty-four healthy male wister rats was randomly divided into two groups, the normal group and the epilepsy group. The Racine grade scores of the two groups of rats were recorded to evaluate the establishment of a rat model of epilepsy. The differentially expressed of microRNAs targeting bcl-2 in the hippocampus of epileptic rats was screened by bioinformatics method. The expression of microRNA21 in hippocampus of epileptic rats was verified by qRT-PCR. Western blot and immunohistochemistry were used to detect the expression of Bcl-2 in rat hippocampus;Hoechst 33258 was used to detect the apoptosis of rat hippocampal neurons. Results: The Racine score of the rats in the epilepsy group was significantly higher than that in the normal control group (P<0.01). Compared with the normal group, the expression of miRNA21 in the hippocampus of the epileptic group increased (P<0.01) and the expression of Bcl-2 decreased (P<0.01). Conclusion: Epilepsy can up-regulate the expression of miRNA21 in rat hippocampal neurons, and it may further induce apoptosis of hippocampal neurons by down-regulating Bcl-2, which may affect epileptic rats.展开更多
Hippocampal neurons undergo various forms of cell death after status epilepticus.Necrostatin-1 specifically inhibits necroptosis mediated by receptor interacting protein kinase 1 (RIP1) and RIP3 receptors.However,ther...Hippocampal neurons undergo various forms of cell death after status epilepticus.Necrostatin-1 specifically inhibits necroptosis mediated by receptor interacting protein kinase 1 (RIP1) and RIP3 receptors.However,there are no reports of necroptosis in mouse models of status epilepticus.Therefore,in this study,we investigated the effects of necrostatin-1 on hippocampal neurons in mice with status epilepticus,and,furthermore,we tested different amounts of the compound to identify the optimal concentration for inhibiting necroptosis and apoptosis.A mouse model of status epilepticus was produced by intraperitoneal injection of kainic acid,12 mg/kg.Different concentrations of necrostatin- 1 (10,20,40,and 80 μM) were administered into the lateral ventricle 15 minutes before kainic acid injection.Hippocampal damage was assessed by hematoxylin-eosin staining 24 hours after the model was successfully produced.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining,western blot assay and immunohistochemistry were used to evaluate the expression of apoptosis-related and necroptosis-related proteins.Necrostatin-1 alleviated damage to hippocampal tissue in the mouse model of epilepsy.The 40 μM concentration of necrostatin-1 significantly decreased the number of apoptotic cells in the hippocampal CA1 region.Furthermore,necrostatin-1 significantly downregulated necroptosis-related proteins (MLKL,RIP1,and RIP3) and apoptosis-related proteins (cleaved-Caspase-3,Bax),and it upregulated the expression of anti-apoptotic protein Bcl-2.Taken together,our findings show that necrostatin-1 effectively inhibits necroptosis and apoptosis in mice with status epilepticus,with the 40 μM concentration of the compound having an optimal effect.The experiments were approved by the Animal Ethics Committee of Fujian Medical University,China (approval No.2016-032) on November 9,2016.展开更多
基金Guangxi Natural Science Foundation(No.2020GXNSFAA238007)Guangxi High-level Innovation Team and Outstanding Scholars Program[Guangxi Higher Education Talents(2020)No.6]+2 种基金Academic Team Building Project of the First Affiliated Hospital of Guangxi TCM University(NO:Courtyard Word[2018]No.146)Guangxi Clinical Research Center of Traditional Chinese Medicine encephalopathy(NO.AD20238028)Guangxi Traditional Chinese Medicine Key Discipline Construction Project(No.GZXK-Z-20-13)。
文摘Objective:To explore the related mechanism of miR-34a affecting epilepsy.Methods:MicroRNA microarray chip and RT-PCR were used to analyze the expression of miR-34a in epileptic rats,and observe the relationship between miR-34a and seizure frequency and EEG.Predicted the target gene of miR-34a,used The dual-luciferase reporter gene assay was used to observe its binding to the target gene of miR-34a,the effect of overexpression and inhibition of miR-34a on the target gene,the effects of overexpression and inhibition of target genes on downstream factors.Results:Microarray chip and RT-PCR detection showed that miR-34a was highly expressed in epilepsy(P<0.05),and miR-34a was positively correlated with seizure frequency(R=0.783,P=0.003),and inhibition of miR-34a inhibited the seizure frequency of epilepsy(P<0.05).The online databases TargetScan and miRDB show that miR-34a and BCL-2 have a predicted binding region,and the dual-luciferase reporter gene experiment proves that the two have a binding relationship.When miR-34a was overexpressed,BCL-2 expression was down-regulated,and conversely,when miR-34a was inhibited,BCL-2 expression was up-regulated.NLRP1 and downstream related factors Caspase-3,Caspase-1,IL-1βand IL-18 were highly expressed in the brain tissue of epileptic rats(P<0.05).When BCL-2 was inhibited,the expression levels of NLRP1 and downstream related factors were increased,and when BCL-2 was inhibited,the result was the opposite.Conclusion:The highly expressed miR-34a promotes epilepsy by inhibiting BCL-2 and activating the NLRP1 inflammasome.
基金Grant from Research Funds of Education Department of Guangxi Zhuang Autonomous Region, No. 200810LX340 Youjiang Medical College for Nationalities, No. 0802002
文摘We studied the effects of Ganoderma lucidum spore powder on Bax and Bcl-2 expression and neuronal apoptosis in pentylenetetrazole-kindled epileptic rats. Sixty adult rats were randomly divided into a control group, an epileptic group （kindled） and three medication groups （150,300 450 mg/kg given to kindled rats）. Bax and Bcl-2 immunohistochemistry and TUNEL labeling showed that the number of Bax- and TUNEL-positive cells in the hippocampus and cerebral cortex decreased significantly in the high-dose medication group, while the number of Bcl-2 immunoreactive cells increased. The Morris water maze test showed that high-dose treatment significantly shortened escape latency and increased spatial probe trial performance. Our findings indicate that a high dose of Ganoderma lucidum spore powder upregulates the expression of antiapoptotic Bcl-2 protein in the hippocampus and cerebral cortex, inhibits proapoptotic Bax expression, and decreases seizure-induced neuronal apoptosis. Further, Ganoderma lucidum appears to protect against epilepsy-related learning and memory impairments.
文摘Objective: To investigate the expression of miRNA21 in hippocampus of rats with pentylenetetrazol-induced epilepsy and its possible mechanism. Methods: Twenty-four healthy male wister rats was randomly divided into two groups, the normal group and the epilepsy group. The Racine grade scores of the two groups of rats were recorded to evaluate the establishment of a rat model of epilepsy. The differentially expressed of microRNAs targeting bcl-2 in the hippocampus of epileptic rats was screened by bioinformatics method. The expression of microRNA21 in hippocampus of epileptic rats was verified by qRT-PCR. Western blot and immunohistochemistry were used to detect the expression of Bcl-2 in rat hippocampus;Hoechst 33258 was used to detect the apoptosis of rat hippocampal neurons. Results: The Racine score of the rats in the epilepsy group was significantly higher than that in the normal control group (P<0.01). Compared with the normal group, the expression of miRNA21 in the hippocampus of the epileptic group increased (P<0.01) and the expression of Bcl-2 decreased (P<0.01). Conclusion: Epilepsy can up-regulate the expression of miRNA21 in rat hippocampal neurons, and it may further induce apoptosis of hippocampal neurons by down-regulating Bcl-2, which may affect epileptic rats.
基金supported by the Key Discipline Construction Project of the Union Hospital of Fujian Province,China,No.Δ211002#
文摘Hippocampal neurons undergo various forms of cell death after status epilepticus.Necrostatin-1 specifically inhibits necroptosis mediated by receptor interacting protein kinase 1 (RIP1) and RIP3 receptors.However,there are no reports of necroptosis in mouse models of status epilepticus.Therefore,in this study,we investigated the effects of necrostatin-1 on hippocampal neurons in mice with status epilepticus,and,furthermore,we tested different amounts of the compound to identify the optimal concentration for inhibiting necroptosis and apoptosis.A mouse model of status epilepticus was produced by intraperitoneal injection of kainic acid,12 mg/kg.Different concentrations of necrostatin- 1 (10,20,40,and 80 μM) were administered into the lateral ventricle 15 minutes before kainic acid injection.Hippocampal damage was assessed by hematoxylin-eosin staining 24 hours after the model was successfully produced.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining,western blot assay and immunohistochemistry were used to evaluate the expression of apoptosis-related and necroptosis-related proteins.Necrostatin-1 alleviated damage to hippocampal tissue in the mouse model of epilepsy.The 40 μM concentration of necrostatin-1 significantly decreased the number of apoptotic cells in the hippocampal CA1 region.Furthermore,necrostatin-1 significantly downregulated necroptosis-related proteins (MLKL,RIP1,and RIP3) and apoptosis-related proteins (cleaved-Caspase-3,Bax),and it upregulated the expression of anti-apoptotic protein Bcl-2.Taken together,our findings show that necrostatin-1 effectively inhibits necroptosis and apoptosis in mice with status epilepticus,with the 40 μM concentration of the compound having an optimal effect.The experiments were approved by the Animal Ethics Committee of Fujian Medical University,China (approval No.2016-032) on November 9,2016.
