Objective:To explore the role of NLRP3/Caspase-1-mediated pyroptosis on peripheral blood monocytes in patients with essential hypertension(EH),and its regulation by pannexin-1(Panx-1)hemi-channels.Methods:The peripher...Objective:To explore the role of NLRP3/Caspase-1-mediated pyroptosis on peripheral blood monocytes in patients with essential hypertension(EH),and its regulation by pannexin-1(Panx-1)hemi-channels.Methods:The peripheral blood of EH patients and healthy subjects was collected,peripheral plasma of the two groups was subsequently separated,and IL-1βcontent in peripheral plasma was further measured by ELISA.In the meantime,the monocytes from peripheral blood were sorted by immunomagnetic beads,and mRNA and protein expression of Panx-1,NLRP3 inflammasome related molecules(NLRP3,ASC,Caspase-1),downstream effector IL-1β,and pyroptosis-related protein GSDMD,were analyzed by RT-qPCR and Western blot,respectively.Subsequently,human primary monocytes in two group were cultured in vitro.Immunofluorescence assay was performed to detect the expression and location of Panx-1 on monocytes.Finally,the cultured monocytes from EH patients were exposed to NLRP3 inflammasome activator lipopolysaccharide(LPS),and also to LPS pretreated with Panx-1 hemi-channel inhibitor probenecid or specific Panx-1 siRNA,followed by collection of the cell culture supernatant and monocytes.The cell viability of each group was detected by CCK-8 assay,IL-1βcontent in the culture supernatant was analyzed by ELISA,and the expression of target proteins on monocytes was determined by Western blot assay.Results:Compared with healthy subjects,EH patients showed higher IL-1βcontent in peripheral plasma,and increased expression in both mRNA and protein levels for Panx-1,NLRP3 inflammasome related molecules,IL-1βand GSDMD on peripheral monocytes.Furthermore,Panx-1 protein in EH patients was significantly higher than that in healthy subjects and localized on monocyte membrane as evidenced by immunofluorescence assay.In the cultured human primary monocytes from EH patients,the cell viability in each group showed no significant difference.LPS stimulation can induce NLRP3-dependent pyroptosis,by showing an increase in IL-1βcontent in cell culture supernatant and protein expressions of NLRP3 inflammasome related molecules and GSDMD on monocytes,by LPS exposure.However,the above stimulating effect of LPS could be counteracted by pretreatment with probenecid or Panx-1 siRNA.Conclusion:The NLRP3/Caspase-1-mediated pyroptosis is activated on peripheral blood monocytes in patients with EH,and this activation could be regulated by Panx-1 hemi-channels.展开更多
目的:观察厄贝沙坦在糖尿病大鼠心肌损伤中发挥的作用,并探讨其对丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPKs)信号通路及其相关因子表达影响。方法:雄性SD大鼠50只,随机分为糖尿病4周(DM4W)组、糖尿病8周(DM8W)组、糖...目的:观察厄贝沙坦在糖尿病大鼠心肌损伤中发挥的作用,并探讨其对丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPKs)信号通路及其相关因子表达影响。方法:雄性SD大鼠50只,随机分为糖尿病4周(DM4W)组、糖尿病8周(DM8W)组、糖尿病8周+厄贝沙坦(DM8W+Ir)组、对照(Con)组、高糖高胆固醇饮食(HC)组,各10只。复制2型糖尿病大鼠模型成功后,DM8W+Ir组给予厄贝沙坦溶液灌胃干预;饲喂第4周时,处死DM4W组大鼠并留取心脏组织标本;饲喂8周后,处死剩余组大鼠。比较各组大鼠空腹血糖、体质量、心体比和左心室质量指数;取大鼠离体心肌行Masson染色观察心肌细胞纤维化改变;ELISA法检测心肌细胞内炎症细胞因子白细胞介素(interleukin,IL)-1β、IL-6、IL-1表达水平;Western blotting法检测心肌组织丝裂原活化蛋白激酶磷酸酶-1(mitogen-activated protein kinase phosphatase-1,MKP-1)、P38丝裂原激活蛋白激酶(P38 mitogen-activated protein kinase,P38MAPK)、细胞外信号调节激酶(extracellular signal-related kinases,P-ERK1/2)蛋白表达水平。结果:HC组大鼠空腹血糖、体质量、心体比、左心室质量指数与Con组差异均无统计学意义(P>0.05),IL-1β、IL-6、IL-1表达水平和MKP-1、P38MAPK、P-ERK1/2蛋白表达与Con组差异亦均无统计学意义(P>0.05)。与Con组、HC组相比,各糖尿病组大鼠Masson染色心肌细胞明显纤维化改变;体质量减轻,心肌细胞内炎症细胞因子IL-1β、IL-6、IL-1表达水平升高,心肌组织MKP-1蛋白表达降低,P38MAPK、P-ERK1/2蛋白表达升高(P<0.05~P<0.01);与DM4W、DM8W组比较,DM8W+Ir组大鼠IL-1β、IL-6、IL-1表达水平降低,MKP-1蛋白表达升高,P38MAPK、P-ERK1/2表达蛋白降低(P<0.05~P<0.01)。结论:厄贝沙坦通过调控MAPKs信号通路发挥改善糖尿病心肌损伤的作用。展开更多
目的:探究血清组蛋白去乙酰化酶3(histone deacetylase 3,HDAC3)、高迁移率蛋白B1(high mobility group B 1,HMGB-1)与非瓣膜性心房颤动发生、发展及结构重构的相关性。方法:选取入院非瓣膜性心房颤动病人为房颤组(atrial fibrillation ...目的:探究血清组蛋白去乙酰化酶3(histone deacetylase 3,HDAC3)、高迁移率蛋白B1(high mobility group B 1,HMGB-1)与非瓣膜性心房颤动发生、发展及结构重构的相关性。方法:选取入院非瓣膜性心房颤动病人为房颤组(atrial fibrillation AF group,n=166),依据相关指南分为阵发性心房颤动组(paroxysmal AF group,n=42)、持续性心房颤动组(persistent AF group,n=63)、永久性心房颤动组(permanent AF group,n=61)。选取同期本院体检50例窦性心律者为对照组,统计各组一般临床资料、心脏彩超、NT-proBNP等结果,采用酶联吸附免疫法(ELISA)测定各组HDAC3、HMGB-1的水平。结果:房颤组HDAC3、HMGB-1、LAD、LVEDD水平均高于对照组(P<0.05);永久性房颤组LAD、LVEDD、HDAC3、HMGB-1水平均高于持续性房颤组与阵发性房颤组(P<0.05),且持续性房颤组较阵发性房颤组亦升高(P<0.05);经Pearson相关分析,HDAC3与LAD(r=0.635,P<0.05)、LVEDD(r=0.651,P<0.05)、HMGB-1(r=0.757,P<0.05)呈正相关;HMGB-1与LAD(r=0.658,P<0.05)、LVEDD(r=0.569,P<0.05)呈正相关;二元logistic回归分析,HDAC3(OR=1.712,95%CI:1.070~2.739,P<0.05)、HMGB-1(OR=1.287,95%CI:1.008~1.634,P<0.05)与心房颤动的发生相关。结论:HDAC3、HMGB-1水平与心房颤动的发生及发展有一定相关性,可能是心房颤动发生的独立危险因素;HDAC3、HMGB-1水平可能与心房颤动心肌结构重构相关。展开更多
基金National Natural Science Foundation of China(No.81970313)Natural Science Foundation of Anhui Province(No.2008085MH238)+1 种基金512 Talent Cultivation Plan of Bengbu Medical College(No.by51201321)Key Program of Natural Science Foundation of Bengbu Medical College(No.2020byzd109)。
文摘Objective:To explore the role of NLRP3/Caspase-1-mediated pyroptosis on peripheral blood monocytes in patients with essential hypertension(EH),and its regulation by pannexin-1(Panx-1)hemi-channels.Methods:The peripheral blood of EH patients and healthy subjects was collected,peripheral plasma of the two groups was subsequently separated,and IL-1βcontent in peripheral plasma was further measured by ELISA.In the meantime,the monocytes from peripheral blood were sorted by immunomagnetic beads,and mRNA and protein expression of Panx-1,NLRP3 inflammasome related molecules(NLRP3,ASC,Caspase-1),downstream effector IL-1β,and pyroptosis-related protein GSDMD,were analyzed by RT-qPCR and Western blot,respectively.Subsequently,human primary monocytes in two group were cultured in vitro.Immunofluorescence assay was performed to detect the expression and location of Panx-1 on monocytes.Finally,the cultured monocytes from EH patients were exposed to NLRP3 inflammasome activator lipopolysaccharide(LPS),and also to LPS pretreated with Panx-1 hemi-channel inhibitor probenecid or specific Panx-1 siRNA,followed by collection of the cell culture supernatant and monocytes.The cell viability of each group was detected by CCK-8 assay,IL-1βcontent in the culture supernatant was analyzed by ELISA,and the expression of target proteins on monocytes was determined by Western blot assay.Results:Compared with healthy subjects,EH patients showed higher IL-1βcontent in peripheral plasma,and increased expression in both mRNA and protein levels for Panx-1,NLRP3 inflammasome related molecules,IL-1βand GSDMD on peripheral monocytes.Furthermore,Panx-1 protein in EH patients was significantly higher than that in healthy subjects and localized on monocyte membrane as evidenced by immunofluorescence assay.In the cultured human primary monocytes from EH patients,the cell viability in each group showed no significant difference.LPS stimulation can induce NLRP3-dependent pyroptosis,by showing an increase in IL-1βcontent in cell culture supernatant and protein expressions of NLRP3 inflammasome related molecules and GSDMD on monocytes,by LPS exposure.However,the above stimulating effect of LPS could be counteracted by pretreatment with probenecid or Panx-1 siRNA.Conclusion:The NLRP3/Caspase-1-mediated pyroptosis is activated on peripheral blood monocytes in patients with EH,and this activation could be regulated by Panx-1 hemi-channels.
文摘目的:观察厄贝沙坦在糖尿病大鼠心肌损伤中发挥的作用,并探讨其对丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPKs)信号通路及其相关因子表达影响。方法:雄性SD大鼠50只,随机分为糖尿病4周(DM4W)组、糖尿病8周(DM8W)组、糖尿病8周+厄贝沙坦(DM8W+Ir)组、对照(Con)组、高糖高胆固醇饮食(HC)组,各10只。复制2型糖尿病大鼠模型成功后,DM8W+Ir组给予厄贝沙坦溶液灌胃干预;饲喂第4周时,处死DM4W组大鼠并留取心脏组织标本;饲喂8周后,处死剩余组大鼠。比较各组大鼠空腹血糖、体质量、心体比和左心室质量指数;取大鼠离体心肌行Masson染色观察心肌细胞纤维化改变;ELISA法检测心肌细胞内炎症细胞因子白细胞介素(interleukin,IL)-1β、IL-6、IL-1表达水平;Western blotting法检测心肌组织丝裂原活化蛋白激酶磷酸酶-1(mitogen-activated protein kinase phosphatase-1,MKP-1)、P38丝裂原激活蛋白激酶(P38 mitogen-activated protein kinase,P38MAPK)、细胞外信号调节激酶(extracellular signal-related kinases,P-ERK1/2)蛋白表达水平。结果:HC组大鼠空腹血糖、体质量、心体比、左心室质量指数与Con组差异均无统计学意义(P>0.05),IL-1β、IL-6、IL-1表达水平和MKP-1、P38MAPK、P-ERK1/2蛋白表达与Con组差异亦均无统计学意义(P>0.05)。与Con组、HC组相比,各糖尿病组大鼠Masson染色心肌细胞明显纤维化改变;体质量减轻,心肌细胞内炎症细胞因子IL-1β、IL-6、IL-1表达水平升高,心肌组织MKP-1蛋白表达降低,P38MAPK、P-ERK1/2蛋白表达升高(P<0.05~P<0.01);与DM4W、DM8W组比较,DM8W+Ir组大鼠IL-1β、IL-6、IL-1表达水平降低,MKP-1蛋白表达升高,P38MAPK、P-ERK1/2表达蛋白降低(P<0.05~P<0.01)。结论:厄贝沙坦通过调控MAPKs信号通路发挥改善糖尿病心肌损伤的作用。
文摘目的:探究血清组蛋白去乙酰化酶3(histone deacetylase 3,HDAC3)、高迁移率蛋白B1(high mobility group B 1,HMGB-1)与非瓣膜性心房颤动发生、发展及结构重构的相关性。方法:选取入院非瓣膜性心房颤动病人为房颤组(atrial fibrillation AF group,n=166),依据相关指南分为阵发性心房颤动组(paroxysmal AF group,n=42)、持续性心房颤动组(persistent AF group,n=63)、永久性心房颤动组(permanent AF group,n=61)。选取同期本院体检50例窦性心律者为对照组,统计各组一般临床资料、心脏彩超、NT-proBNP等结果,采用酶联吸附免疫法(ELISA)测定各组HDAC3、HMGB-1的水平。结果:房颤组HDAC3、HMGB-1、LAD、LVEDD水平均高于对照组(P<0.05);永久性房颤组LAD、LVEDD、HDAC3、HMGB-1水平均高于持续性房颤组与阵发性房颤组(P<0.05),且持续性房颤组较阵发性房颤组亦升高(P<0.05);经Pearson相关分析,HDAC3与LAD(r=0.635,P<0.05)、LVEDD(r=0.651,P<0.05)、HMGB-1(r=0.757,P<0.05)呈正相关;HMGB-1与LAD(r=0.658,P<0.05)、LVEDD(r=0.569,P<0.05)呈正相关;二元logistic回归分析,HDAC3(OR=1.712,95%CI:1.070~2.739,P<0.05)、HMGB-1(OR=1.287,95%CI:1.008~1.634,P<0.05)与心房颤动的发生相关。结论:HDAC3、HMGB-1水平与心房颤动的发生及发展有一定相关性,可能是心房颤动发生的独立危险因素;HDAC3、HMGB-1水平可能与心房颤动心肌结构重构相关。
