目的观察重组人促红细胞生成素(recombinant human erythropoietin,rhEPO)对缺血/再灌注损伤大鼠心肌细胞Mitofusin2(Mfn2)蛋白表达的影响及其抗心肌细胞凋亡的作用。方法选取成年SD大鼠35只,随机分为正常组(Normal),假手术组(Sham),缺...目的观察重组人促红细胞生成素(recombinant human erythropoietin,rhEPO)对缺血/再灌注损伤大鼠心肌细胞Mitofusin2(Mfn2)蛋白表达的影响及其抗心肌细胞凋亡的作用。方法选取成年SD大鼠35只,随机分为正常组(Normal),假手术组(Sham),缺血再灌注组(I/R),缺血再灌注EPO治疗组(I/R+EPO)。各组分别于再灌注3h和24h后,剪取心脏缺血/再灌注损伤区域,用脱氧核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测心肌细胞凋亡,免疫组化法检测Mfn2蛋白的表达。结果再灌注3h和24h后,与正常组和假手术组相比,I/R组Mfn2蛋白的表达和心肌细胞凋亡均显著增加;与I/R组相比,I/R+EPO组Mfn2蛋白的表达和心肌细胞凋亡均显著降低。结论EPO可以下调缺血再灌注损伤后心肌细胞Mfn2蛋白的表达,抑制心肌细胞的凋亡。展开更多
Objective:To identify the molecular mechanisms of the effects of the Buyin Qianzheng formula(BYQZF)on the mitochondrial dynamics in a Parkin overexpression Parkinson's disease(PD)cell model.Methods:First,a stable ...Objective:To identify the molecular mechanisms of the effects of the Buyin Qianzheng formula(BYQZF)on the mitochondrial dynamics in a Parkin overexpression Parkinson's disease(PD)cell model.Methods:First,a stable Parkin overexpression cell model was constructed using plasmid transfection.Then,we examined the protective effect of BYQZF on the mitochondrial dysfunction of the Parkin overexpression PD cell model induced by neurotoxin 1-methyl-4-phenylpyridinium ion(MPPþ).The mRNA expression level of Parkin was evaluated using real-time quantitative PCR.The cell survival rate was detected using the Cell Counting Kit-8 assay.We evaluated the cellular adenosine triphosphate(ATP)levels using luciferase assays.A laser scanning confocal microscope was used to observe the mitochondrial morphology,activity,and mitochondrial membrane potential(DJm).Western blot was conducted to evaluate the levels of the fusion proteins mitofusin1,mitofusin2,optic atrophy 1,dynaminrelated protein 1,and mitochondrial fission protein 1.Results:Parkin overexpression attenuated MPPþ-induced mitochondrial damage,increased mitochondrial activity and DJm.BYQZF increased the survival of MPPþ-induced cells that overexpressed Parkin and upregulated the mitochondrial form factor and activity.It also inhibited a decrease in the DJm and ATP levels.These findings suggested that BYQZF protected against MPPþ-induced mitochondrial dysfunction and enhanced the protective effect of Parkin overexpression.Furthermore,the formula upregulated the expression of the fusion proteins mitofusin1,mitofusin2,and optic atrophy 1(closely related to mitochondrial quality remodeling),and reduced the expression of the fission protein dynamicrelated protein 1,as well as mitochondrial fission protein 1.Conclusion:The mechanism by which BYQZF increased the mitochondrial protective effect of Parkin gene overexpression in MPPþ-induced cells may be related to improving mitochondrial function and regulating the balance of mitochondrial division and fusion proteins.展开更多
目的探讨姜黄素对脓毒症小鼠急性肺损伤的保护作用及机制。方法取120只清洁级BALB/c雄性小鼠随机(随机数字法)分成8组,即假手术组、脓毒症组、姜黄素对照组、姜黄素干预组、阴性病毒-脓毒症组、阴性病毒-姜黄素干预组、线粒体融合蛋白(M...目的探讨姜黄素对脓毒症小鼠急性肺损伤的保护作用及机制。方法取120只清洁级BALB/c雄性小鼠随机(随机数字法)分成8组,即假手术组、脓毒症组、姜黄素对照组、姜黄素干预组、阴性病毒-脓毒症组、阴性病毒-姜黄素干预组、线粒体融合蛋白(Mfn2)干扰-脓毒症组、Mfn2干扰-姜黄素干预组,每组15只。脓毒症组及姜黄素干预组通过盲肠结扎穿孔术(cecal ligation and puncture,CLP)造模,姜黄素干预组及姜黄素对照组予姜黄素200 mg/(kg·d)灌胃1周,阴性病毒-脓毒症组及阴性病毒-姜黄素干预组通过尾静脉注射阴性腺相关病毒来建立,Mfn2干扰-脓毒症组及Mfn2干扰-姜黄素干预组通过尾静脉注射携带Mfn2干扰序列的腺相关病毒建立模型,各组均于24 h后处死小鼠。通过肺湿干比和病理学检查反映肺损伤程度,ELISA法检测肺泡灌洗液的炎症因子TNF-α和IL-6,Western blot检测细胞凋亡的关键分子caspase-3的活化,并通过TUNEL检测细胞凋亡。采用SPSS 22.0软件进行统计分析,计数资料比较采用χ^2检验,计量资料组间比较采用单因素方差分析。结果与假手术组比较,脓毒症组小鼠肺组织湿干重比显著增加(71.11±3.78 vs 31.11±5.61,P=0.002);与假手术组相比,脓毒症组织病理评分明显增加(P=0.006),炎症因子TNF-α(P=0.001)和IL-6(P=0.012)显著增高,肺组织细胞凋亡及凋亡相关蛋白caspase-3 cleaved表达同样明显增加(P=0.001)。与脓毒症组比较,姜黄素干预组小鼠肺组织湿干重比明显降低(32.84±6.15 vs 71.11±3.78,P=0.004),组织病理评分显著降低(P=0.004),炎症因子TNF-α(P=0.013)和IL-6(P=0.003)均明显降低,肺组织细胞凋亡及凋亡相关蛋白caspase-3 cleaved表达同样显著减少(P=0.012)。在下调Mfn2后,Mfn2干扰-姜黄素干预组与Mfn2干扰-脓毒症组相比,小鼠肺组织干湿重比无明显降低,组织病理评分也无显著降低,炎症因子TNF-α和IL-6均无明显降低,肺组织细胞凋亡及凋亡相关蛋白caspase-3 cleaved表达无显著减少。结论姜黄素可能通过上调线粒体融合蛋白2的表达减轻脓毒症急性肺损伤。展开更多
文摘目的观察重组人促红细胞生成素(recombinant human erythropoietin,rhEPO)对缺血/再灌注损伤大鼠心肌细胞Mitofusin2(Mfn2)蛋白表达的影响及其抗心肌细胞凋亡的作用。方法选取成年SD大鼠35只,随机分为正常组(Normal),假手术组(Sham),缺血再灌注组(I/R),缺血再灌注EPO治疗组(I/R+EPO)。各组分别于再灌注3h和24h后,剪取心脏缺血/再灌注损伤区域,用脱氧核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测心肌细胞凋亡,免疫组化法检测Mfn2蛋白的表达。结果再灌注3h和24h后,与正常组和假手术组相比,I/R组Mfn2蛋白的表达和心肌细胞凋亡均显著增加;与I/R组相比,I/R+EPO组Mfn2蛋白的表达和心肌细胞凋亡均显著降低。结论EPO可以下调缺血再灌注损伤后心肌细胞Mfn2蛋白的表达,抑制心肌细胞的凋亡。
基金This work is supported by the National Natural Science Foundation of China(81774110 and 81573773).
文摘Objective:To identify the molecular mechanisms of the effects of the Buyin Qianzheng formula(BYQZF)on the mitochondrial dynamics in a Parkin overexpression Parkinson's disease(PD)cell model.Methods:First,a stable Parkin overexpression cell model was constructed using plasmid transfection.Then,we examined the protective effect of BYQZF on the mitochondrial dysfunction of the Parkin overexpression PD cell model induced by neurotoxin 1-methyl-4-phenylpyridinium ion(MPPþ).The mRNA expression level of Parkin was evaluated using real-time quantitative PCR.The cell survival rate was detected using the Cell Counting Kit-8 assay.We evaluated the cellular adenosine triphosphate(ATP)levels using luciferase assays.A laser scanning confocal microscope was used to observe the mitochondrial morphology,activity,and mitochondrial membrane potential(DJm).Western blot was conducted to evaluate the levels of the fusion proteins mitofusin1,mitofusin2,optic atrophy 1,dynaminrelated protein 1,and mitochondrial fission protein 1.Results:Parkin overexpression attenuated MPPþ-induced mitochondrial damage,increased mitochondrial activity and DJm.BYQZF increased the survival of MPPþ-induced cells that overexpressed Parkin and upregulated the mitochondrial form factor and activity.It also inhibited a decrease in the DJm and ATP levels.These findings suggested that BYQZF protected against MPPþ-induced mitochondrial dysfunction and enhanced the protective effect of Parkin overexpression.Furthermore,the formula upregulated the expression of the fusion proteins mitofusin1,mitofusin2,and optic atrophy 1(closely related to mitochondrial quality remodeling),and reduced the expression of the fission protein dynamicrelated protein 1,as well as mitochondrial fission protein 1.Conclusion:The mechanism by which BYQZF increased the mitochondrial protective effect of Parkin gene overexpression in MPPþ-induced cells may be related to improving mitochondrial function and regulating the balance of mitochondrial division and fusion proteins.
文摘目的探讨姜黄素对脓毒症小鼠急性肺损伤的保护作用及机制。方法取120只清洁级BALB/c雄性小鼠随机(随机数字法)分成8组,即假手术组、脓毒症组、姜黄素对照组、姜黄素干预组、阴性病毒-脓毒症组、阴性病毒-姜黄素干预组、线粒体融合蛋白(Mfn2)干扰-脓毒症组、Mfn2干扰-姜黄素干预组,每组15只。脓毒症组及姜黄素干预组通过盲肠结扎穿孔术(cecal ligation and puncture,CLP)造模,姜黄素干预组及姜黄素对照组予姜黄素200 mg/(kg·d)灌胃1周,阴性病毒-脓毒症组及阴性病毒-姜黄素干预组通过尾静脉注射阴性腺相关病毒来建立,Mfn2干扰-脓毒症组及Mfn2干扰-姜黄素干预组通过尾静脉注射携带Mfn2干扰序列的腺相关病毒建立模型,各组均于24 h后处死小鼠。通过肺湿干比和病理学检查反映肺损伤程度,ELISA法检测肺泡灌洗液的炎症因子TNF-α和IL-6,Western blot检测细胞凋亡的关键分子caspase-3的活化,并通过TUNEL检测细胞凋亡。采用SPSS 22.0软件进行统计分析,计数资料比较采用χ^2检验,计量资料组间比较采用单因素方差分析。结果与假手术组比较,脓毒症组小鼠肺组织湿干重比显著增加(71.11±3.78 vs 31.11±5.61,P=0.002);与假手术组相比,脓毒症组织病理评分明显增加(P=0.006),炎症因子TNF-α(P=0.001)和IL-6(P=0.012)显著增高,肺组织细胞凋亡及凋亡相关蛋白caspase-3 cleaved表达同样明显增加(P=0.001)。与脓毒症组比较,姜黄素干预组小鼠肺组织湿干重比明显降低(32.84±6.15 vs 71.11±3.78,P=0.004),组织病理评分显著降低(P=0.004),炎症因子TNF-α(P=0.013)和IL-6(P=0.003)均明显降低,肺组织细胞凋亡及凋亡相关蛋白caspase-3 cleaved表达同样显著减少(P=0.012)。在下调Mfn2后,Mfn2干扰-姜黄素干预组与Mfn2干扰-脓毒症组相比,小鼠肺组织干湿重比无明显降低,组织病理评分也无显著降低,炎症因子TNF-α和IL-6均无明显降低,肺组织细胞凋亡及凋亡相关蛋白caspase-3 cleaved表达无显著减少。结论姜黄素可能通过上调线粒体融合蛋白2的表达减轻脓毒症急性肺损伤。
