BACKGROUND Hepatocellular carcinoma(HCC)has become a great threat for people’s health.Many long noncoding RNAs are involved in the pathogenesis of HCC.SNHG15,as a tissue specific long noncoding RNAs,has been studied ...BACKGROUND Hepatocellular carcinoma(HCC)has become a great threat for people’s health.Many long noncoding RNAs are involved in the pathogenesis of HCC.SNHG15,as a tissue specific long noncoding RNAs,has been studied in many human cancers,except HCC.AIM To explore the regulatory mechanism of SNHG15 in HCC.METHODS In the present research,101 HCC patient samples,two HCC cell lines and one normal liver cell line were used.RT-qPCR and Western blot analysis were applied to detect SNHG15,miR-490-3p and histone deacetylase 2(HDAC2)expression.The regulatory mechanism of SNHG15 was investigated using CCK-8,Transwell and luciferase reporter assays.RESULTS Our research showed that up-regulation of SNHG15 was found in HCC and was related to aggressive behaviors in HCC patients.Moreover,knockdown of SNHG15 restrained HCC cell proliferation,migration and invasion.In addition,SNHG15 served as a molecular sponge for miR-490-3p.Further,miR-490-3p directly targets HDAC2.HDAC2 was involved in HCC progression by interacting with the SNHG15/miR-490-3p axis.CONCLUSION In conclusion,long noncoding RNA SNHG15 promotes HCC progression by mediating the miR-490-3p/HDAC2 axis in HCC.展开更多
Background:Cochlear hair cell injury is a common pathological feature of hearing loss.The basic helix-loop-helix family,member e40(Bhlhe40),a gene belonging to the basic helix-loop-helix(bHLH)family,exhibits strong tr...Background:Cochlear hair cell injury is a common pathological feature of hearing loss.The basic helix-loop-helix family,member e40(Bhlhe40),a gene belonging to the basic helix-loop-helix(bHLH)family,exhibits strong transcriptional repression activity.Methods:Oxidative damage,in House Ear Institute-Organ of Corti 1(HEI-OC1)cells,was caused using hydrogen peroxide(H2O2).The Ad-Bhlhe40 particles were constructed to overexpress Bhlhe40 in HEI-OC1 cells.Various assays including cell counting kit-8(CCK-8),terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay(TUNEL),flow cytometry,immunofluorescence,and corresponding commercial kits were employed to investigate the impacts of Bhlhe40 on cell viability,apoptosis,oxidative stress levels,mitochondrial membrane potential and cellular senescence.Additionally,a dual-luciferase reporter assay was performed to confirm the targeting of the histone deacetylases 2(Hdac2)by Bhlhe40.Results:The results revealed that Bhlhe40 was downregulated in H_(2)O_(2)-treated HEI-OC1 cells,but its overexpression improved cell viability and mitigated H_(2)O_(2)-induced oxidative injury in HEI-OC1 cells with increase of superoxide dismutase(SOD),catalase(CAT)and glutathione peroxidase(GPx)activities and decrease of reactive oxygen species(ROS)levels.Besides,overexpression of Bhlhe40 suppressed H_(2)O_(2)-triggered cell senescence,as evidenced by the fact that the upregulation of P53,P21,and P16 in HEI-OC1 cells treated with H2O2 were all alleviated by Bhlhe40 overexpression.And we further verified that overexpression of Bhlhe40 could inhibit the expression of Hdac2,which may be related to the repression of Hdac2 transcription.Conclusion:This study suggests that Bhlhe40 plays a protective role against senescence and oxidative damage in cochlear hair cells exposed to H2O2.展开更多
OBJECTIVE To investigate the mechanisms of histone deacetylase 6(HDAC6)deacetylation activity on NALP3 inflammasome activation and explore the protective effect s of pharmacological inhibition of HDAC6 on dopaminergic...OBJECTIVE To investigate the mechanisms of histone deacetylase 6(HDAC6)deacetylation activity on NALP3 inflammasome activation and explore the protective effect s of pharmacological inhibition of HDAC6 on dopaminergic injury.METHODS In vitro and in vivo6-OHDA induced Parkinson disease(PD) model was used.To distinguish the effect of deacetylase catalytic domains of HDAC6,we used a specific HDAC6 inhibitor tubastatin A(TBA),siRNAHDAC6,and pcDNA-HDAC6-FLAG plasmid.First,the role of pharmacological inhibition or siRNA or overexpression of HDAC6 on NALP3 inflammasome and cell death was explored by using Western blotting,TUNEL,and flow cytometric analysis.Then,the acetylation level of peroxiredoxin 2(Prx2) and the production of reactive oxygen species(ROS) in cells under different treatments was examined by using immunoprecipitation and DCFH-DA fluorescence assay.The effects of TBA on neuroinflammation and nigrostriatal dopaminergic system in vivo was further investigated by using Western blotting,immunohistochemistry and HPLC analysis.RESULTS TBA remarkably inhibited 6-OHDA induced NALP3 inflammasome activation,reduced dopaminergic neurodegeneration and neuroinflammation as demonstrated by increased TH-positive neurons,striatal levels of DA and its metabolites,and decreased gliocyte proliferation.TBA recovered acetylation of Prx2,and reduced ROS production,which was associated with decreased NALP3 inflammasome activation.CONCLUSION HDAC6 may medicate deacetylation of Prx2 contributes to NALP3 inflammasome activation in PD pathology,suggesting that the development of specific pharmacological inhibitors of HDAC6 be required for this kind of disease.展开更多
目的观察参芪补肺汤对慢性阻塞性肺疾病(COPD)肺气虚证大鼠支气管平滑肌(ASM)中乙酰化组蛋白H4、组蛋白去乙酰化酶-2(HDAC2)和核因子-κB p65(NF-κB p65)表达的影响。方法取SD大鼠40只,随机分为正常组、模型组、参芪补肺汤组、氨茶碱组...目的观察参芪补肺汤对慢性阻塞性肺疾病(COPD)肺气虚证大鼠支气管平滑肌(ASM)中乙酰化组蛋白H4、组蛋白去乙酰化酶-2(HDAC2)和核因子-κB p65(NF-κB p65)表达的影响。方法取SD大鼠40只,随机分为正常组、模型组、参芪补肺汤组、氨茶碱组,每组10只。运用气管内注射脂多糖加烟熏28 d的方法建立COPD肺气虚证大鼠模型。光镜下观察肺组织的病理形态学变化,运用图像分析法测量小气道管壁和ASM的厚度,采用免疫组化和Western blot方法检测大鼠ASM中乙酰化组蛋白H4、HDAC2和NF-κB p65的蛋白表达,实时荧光定量PCR方法检测大鼠ASM组织中HDAC2 m RNA和NF-κB p65 m RNA的表达。结果与正常组比较,模型组大鼠气道管壁和ASM厚度明显增高(P<0.05);与模型组比较,参芪补肺汤组和氨茶碱组气道管壁和ASM厚度明显降低(P<0.05);参芪补肺汤组与氨茶碱组比较差异无统计学意义(P>0.05)。与正常组比较,模型组乙酰化组蛋白H4的蛋白表达、NF-κB p65 m RNA和蛋白的表达明显增高(P<0.05);HDAC2m RNA和蛋白的表达均明显降低(P<0.05);与模型组比较,参芪补肺汤组和氨茶碱组乙酰化组蛋白H4的蛋白表达、NF-κB p65 m RNA和蛋白的表达明显增高(P<0.05);HDAC2 m RNA和蛋白的表达明显降低(P<0.05)。参芪补肺汤组与氨茶碱组比较差异均无统计学意义(P>0.05)。结论参芪补肺汤可抑制COPD肺气虚证模型大鼠ASM增殖,其机制与其提高HDAC2的表达,使组蛋白H4去乙酰化,从而抑制NF-κB p65的表达有关。展开更多
文摘BACKGROUND Hepatocellular carcinoma(HCC)has become a great threat for people’s health.Many long noncoding RNAs are involved in the pathogenesis of HCC.SNHG15,as a tissue specific long noncoding RNAs,has been studied in many human cancers,except HCC.AIM To explore the regulatory mechanism of SNHG15 in HCC.METHODS In the present research,101 HCC patient samples,two HCC cell lines and one normal liver cell line were used.RT-qPCR and Western blot analysis were applied to detect SNHG15,miR-490-3p and histone deacetylase 2(HDAC2)expression.The regulatory mechanism of SNHG15 was investigated using CCK-8,Transwell and luciferase reporter assays.RESULTS Our research showed that up-regulation of SNHG15 was found in HCC and was related to aggressive behaviors in HCC patients.Moreover,knockdown of SNHG15 restrained HCC cell proliferation,migration and invasion.In addition,SNHG15 served as a molecular sponge for miR-490-3p.Further,miR-490-3p directly targets HDAC2.HDAC2 was involved in HCC progression by interacting with the SNHG15/miR-490-3p axis.CONCLUSION In conclusion,long noncoding RNA SNHG15 promotes HCC progression by mediating the miR-490-3p/HDAC2 axis in HCC.
基金This research was supported by the Special Fund for Economic and Technological Development of Longgang District,Shenzhen(LGKCYLWS2021000030).
文摘Background:Cochlear hair cell injury is a common pathological feature of hearing loss.The basic helix-loop-helix family,member e40(Bhlhe40),a gene belonging to the basic helix-loop-helix(bHLH)family,exhibits strong transcriptional repression activity.Methods:Oxidative damage,in House Ear Institute-Organ of Corti 1(HEI-OC1)cells,was caused using hydrogen peroxide(H2O2).The Ad-Bhlhe40 particles were constructed to overexpress Bhlhe40 in HEI-OC1 cells.Various assays including cell counting kit-8(CCK-8),terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay(TUNEL),flow cytometry,immunofluorescence,and corresponding commercial kits were employed to investigate the impacts of Bhlhe40 on cell viability,apoptosis,oxidative stress levels,mitochondrial membrane potential and cellular senescence.Additionally,a dual-luciferase reporter assay was performed to confirm the targeting of the histone deacetylases 2(Hdac2)by Bhlhe40.Results:The results revealed that Bhlhe40 was downregulated in H_(2)O_(2)-treated HEI-OC1 cells,but its overexpression improved cell viability and mitigated H_(2)O_(2)-induced oxidative injury in HEI-OC1 cells with increase of superoxide dismutase(SOD),catalase(CAT)and glutathione peroxidase(GPx)activities and decrease of reactive oxygen species(ROS)levels.Besides,overexpression of Bhlhe40 suppressed H_(2)O_(2)-triggered cell senescence,as evidenced by the fact that the upregulation of P53,P21,and P16 in HEI-OC1 cells treated with H2O2 were all alleviated by Bhlhe40 overexpression.And we further verified that overexpression of Bhlhe40 could inhibit the expression of Hdac2,which may be related to the repression of Hdac2 transcription.Conclusion:This study suggests that Bhlhe40 plays a protective role against senescence and oxidative damage in cochlear hair cells exposed to H2O2.
基金Key Technologies Research and Development Program of Shandong Province (2017GSF18171,2018GSF118139).
文摘OBJECTIVE To investigate the mechanisms of histone deacetylase 6(HDAC6)deacetylation activity on NALP3 inflammasome activation and explore the protective effect s of pharmacological inhibition of HDAC6 on dopaminergic injury.METHODS In vitro and in vivo6-OHDA induced Parkinson disease(PD) model was used.To distinguish the effect of deacetylase catalytic domains of HDAC6,we used a specific HDAC6 inhibitor tubastatin A(TBA),siRNAHDAC6,and pcDNA-HDAC6-FLAG plasmid.First,the role of pharmacological inhibition or siRNA or overexpression of HDAC6 on NALP3 inflammasome and cell death was explored by using Western blotting,TUNEL,and flow cytometric analysis.Then,the acetylation level of peroxiredoxin 2(Prx2) and the production of reactive oxygen species(ROS) in cells under different treatments was examined by using immunoprecipitation and DCFH-DA fluorescence assay.The effects of TBA on neuroinflammation and nigrostriatal dopaminergic system in vivo was further investigated by using Western blotting,immunohistochemistry and HPLC analysis.RESULTS TBA remarkably inhibited 6-OHDA induced NALP3 inflammasome activation,reduced dopaminergic neurodegeneration and neuroinflammation as demonstrated by increased TH-positive neurons,striatal levels of DA and its metabolites,and decreased gliocyte proliferation.TBA recovered acetylation of Prx2,and reduced ROS production,which was associated with decreased NALP3 inflammasome activation.CONCLUSION HDAC6 may medicate deacetylation of Prx2 contributes to NALP3 inflammasome activation in PD pathology,suggesting that the development of specific pharmacological inhibitors of HDAC6 be required for this kind of disease.
文摘目的观察参芪补肺汤对慢性阻塞性肺疾病(COPD)肺气虚证大鼠支气管平滑肌(ASM)中乙酰化组蛋白H4、组蛋白去乙酰化酶-2(HDAC2)和核因子-κB p65(NF-κB p65)表达的影响。方法取SD大鼠40只,随机分为正常组、模型组、参芪补肺汤组、氨茶碱组,每组10只。运用气管内注射脂多糖加烟熏28 d的方法建立COPD肺气虚证大鼠模型。光镜下观察肺组织的病理形态学变化,运用图像分析法测量小气道管壁和ASM的厚度,采用免疫组化和Western blot方法检测大鼠ASM中乙酰化组蛋白H4、HDAC2和NF-κB p65的蛋白表达,实时荧光定量PCR方法检测大鼠ASM组织中HDAC2 m RNA和NF-κB p65 m RNA的表达。结果与正常组比较,模型组大鼠气道管壁和ASM厚度明显增高(P<0.05);与模型组比较,参芪补肺汤组和氨茶碱组气道管壁和ASM厚度明显降低(P<0.05);参芪补肺汤组与氨茶碱组比较差异无统计学意义(P>0.05)。与正常组比较,模型组乙酰化组蛋白H4的蛋白表达、NF-κB p65 m RNA和蛋白的表达明显增高(P<0.05);HDAC2m RNA和蛋白的表达均明显降低(P<0.05);与模型组比较,参芪补肺汤组和氨茶碱组乙酰化组蛋白H4的蛋白表达、NF-κB p65 m RNA和蛋白的表达明显增高(P<0.05);HDAC2 m RNA和蛋白的表达明显降低(P<0.05)。参芪补肺汤组与氨茶碱组比较差异均无统计学意义(P>0.05)。结论参芪补肺汤可抑制COPD肺气虚证模型大鼠ASM增殖,其机制与其提高HDAC2的表达,使组蛋白H4去乙酰化,从而抑制NF-κB p65的表达有关。
