Objective Stroke is a leading cause of death and disability worldwide,with ischemic stroke accounting for 80%-85%of cases.Despite the prevalence,effective treatments remain scarce.The compelling evidence suggest that ...Objective Stroke is a leading cause of death and disability worldwide,with ischemic stroke accounting for 80%-85%of cases.Despite the prevalence,effective treatments remain scarce.The compelling evidence suggest that high concentrations of ATP in the brain post-stroke can trigger irreversible neuronal damage and necrosis,contributing to a range of neurocellular dysfunctions.Pyroptosis,a recently identified form of programmed cell death,is characterized by caspase-1 activation and the action of the Gasdermin D(GSDMD)protein family,leading to cell perforation and inflammatory death.Methods In this study,human neuroblastoma SH-SY5Y cells were used to investigate the mechanisms of ATP-induced neurotoxicity and the protective effects of hydrogen sulfide(H_(2)S)against this toxicity through the antagonization of pyroptosis.We employed CCK-8 and LDH assays to assess cell viability.YO-PRO-1 fluorescent dyes and flow cytometry were conducted for detecting changes in cell membrane permeability.Western blot analysis was used to measure protein levels associated with cellular dysfunction.Results Our results indicate that high concentrations of ATP enhance cytotoxicity and increase cell membrane permeability in SH-SY5Y cells,that are mitigated by the H_(2)S donor NaHS.Furthermore,ATP was found to promote the activation of the NOD-like receptor pyrin domain-containing 1(NLRP-1),caspase-1,and the cleavage of GSDMD,with NaHS significantly attenuating these effects.Conclusion Our research suggests that H2S protects SH-SY5Y cells from ATP-induced neurotoxicity through a mechanism mediated by the NLRP1,caspase-1,and GSDMD pathway.展开更多
目的脑信号蛋白4D(Sema4D)是发现最早的免疫信号素,可能参与骨质疏松症的发生和发展。文章通过检测骨质疏松小鼠血清中可溶性脑信号蛋白4D(sSema4D)的水平,探寻sSema4D与骨质疏松症发病的关系。方法选取3月龄SPF级野生型C57小鼠,随机抽...目的脑信号蛋白4D(Sema4D)是发现最早的免疫信号素,可能参与骨质疏松症的发生和发展。文章通过检测骨质疏松小鼠血清中可溶性脑信号蛋白4D(sSema4D)的水平,探寻sSema4D与骨质疏松症发病的关系。方法选取3月龄SPF级野生型C57小鼠,随机抽样法分为两组:骨质疏松组(n=50):每天给予0.9%维甲酸(90 mL/kg)灌胃共21 d;对照组(n=50):每天灌以0.9%氯化钠注射液(90 mL/kg)。应用Micro-CT对小鼠胫骨近端干骺端进行扫描并行三维重建,观察骨小梁数目、结构、形态、粗细、排列及走向情况;使用Inveon Research Work Place软件对胫骨近端干骺端兴趣区域的CT值进行处理,测量小鼠骨小梁数目、骨小梁厚度、骨小梁体积分数、骨小梁分离度。采用硬组织切片技术切片,苦味酸-酸性品红法染色,显微镜下观察股骨、胫骨组织骨胶原纤维病理学改变。采用酶联免疫吸附法检测小鼠血清中sSema4D水平。结果骨质疏松症组小鼠的骨小梁数目、骨小梁厚度、骨小梁体积分数较对照组明显减少(5.8 mm vs 7.0 mm,0.06 mm vs 0.08 mm,0.3%vs0.5%,P<0.01),骨小梁分离度明显增大[(0.69±0.13)mm vs(0.54±0.12)mm,P<0.05]。骨组织病理学表现:苦味酸-酸性品红法染色镜下显示骨质疏松组小鼠的胶原纤维排列疏松、稀少,纤维结构紊乱,部分胶原纤维连续性变差,溶解及断裂,骨髓腔增大等;血清中sSema4D水平:骨质疏松症组小鼠血清sSema4D较对照组的sSema4D显著升高(P<0.01)。结论 sSema4D可能促使骨质疏松症的发生。展开更多
文摘Objective Stroke is a leading cause of death and disability worldwide,with ischemic stroke accounting for 80%-85%of cases.Despite the prevalence,effective treatments remain scarce.The compelling evidence suggest that high concentrations of ATP in the brain post-stroke can trigger irreversible neuronal damage and necrosis,contributing to a range of neurocellular dysfunctions.Pyroptosis,a recently identified form of programmed cell death,is characterized by caspase-1 activation and the action of the Gasdermin D(GSDMD)protein family,leading to cell perforation and inflammatory death.Methods In this study,human neuroblastoma SH-SY5Y cells were used to investigate the mechanisms of ATP-induced neurotoxicity and the protective effects of hydrogen sulfide(H_(2)S)against this toxicity through the antagonization of pyroptosis.We employed CCK-8 and LDH assays to assess cell viability.YO-PRO-1 fluorescent dyes and flow cytometry were conducted for detecting changes in cell membrane permeability.Western blot analysis was used to measure protein levels associated with cellular dysfunction.Results Our results indicate that high concentrations of ATP enhance cytotoxicity and increase cell membrane permeability in SH-SY5Y cells,that are mitigated by the H_(2)S donor NaHS.Furthermore,ATP was found to promote the activation of the NOD-like receptor pyrin domain-containing 1(NLRP-1),caspase-1,and the cleavage of GSDMD,with NaHS significantly attenuating these effects.Conclusion Our research suggests that H2S protects SH-SY5Y cells from ATP-induced neurotoxicity through a mechanism mediated by the NLRP1,caspase-1,and GSDMD pathway.
文摘目的脑信号蛋白4D(Sema4D)是发现最早的免疫信号素,可能参与骨质疏松症的发生和发展。文章通过检测骨质疏松小鼠血清中可溶性脑信号蛋白4D(sSema4D)的水平,探寻sSema4D与骨质疏松症发病的关系。方法选取3月龄SPF级野生型C57小鼠,随机抽样法分为两组:骨质疏松组(n=50):每天给予0.9%维甲酸(90 mL/kg)灌胃共21 d;对照组(n=50):每天灌以0.9%氯化钠注射液(90 mL/kg)。应用Micro-CT对小鼠胫骨近端干骺端进行扫描并行三维重建,观察骨小梁数目、结构、形态、粗细、排列及走向情况;使用Inveon Research Work Place软件对胫骨近端干骺端兴趣区域的CT值进行处理,测量小鼠骨小梁数目、骨小梁厚度、骨小梁体积分数、骨小梁分离度。采用硬组织切片技术切片,苦味酸-酸性品红法染色,显微镜下观察股骨、胫骨组织骨胶原纤维病理学改变。采用酶联免疫吸附法检测小鼠血清中sSema4D水平。结果骨质疏松症组小鼠的骨小梁数目、骨小梁厚度、骨小梁体积分数较对照组明显减少(5.8 mm vs 7.0 mm,0.06 mm vs 0.08 mm,0.3%vs0.5%,P<0.01),骨小梁分离度明显增大[(0.69±0.13)mm vs(0.54±0.12)mm,P<0.05]。骨组织病理学表现:苦味酸-酸性品红法染色镜下显示骨质疏松组小鼠的胶原纤维排列疏松、稀少,纤维结构紊乱,部分胶原纤维连续性变差,溶解及断裂,骨髓腔增大等;血清中sSema4D水平:骨质疏松症组小鼠血清sSema4D较对照组的sSema4D显著升高(P<0.01)。结论 sSema4D可能促使骨质疏松症的发生。
