OBJECTIVE Neocryptotanshinone(NCTS)is a natural product extracted from traditional Chinese herb Salvia miltiorrhiza Bunge.Previous studies have demonstrated the anti-inflammatory of NCTS in lipopolysaccharide(LPS)-sti...OBJECTIVE Neocryptotanshinone(NCTS)is a natural product extracted from traditional Chinese herb Salvia miltiorrhiza Bunge.Previous studies have demonstrated the anti-inflammatory of NCTS in lipopolysaccharide(LPS)-stimulated mouse macrophage(RAW 264.7).However,the protective effect and mechanism of NCTS in cardiomy⁃ocytes are still undefined.This study is to investigate whether NCTS exerts its cardioprotective effect against hypoxia/re⁃oxygenation(H/R)-induced H9C2 cell injury.METHODS The model of H/R injury was established through hypoxia for 8 h and reoxygenation for 12 h in H9C2 cardiomyocytes of rats.Cultured cardiomyocytes were randomly divided into four groups,control group,H/R group,H/R+NCTS pretreated group(1,2,5 and 10μmol·L^-1),and H/R+NCTS+HX531(an RXRαantagonist,2μmol·L^-1)co-treated group.The cell viability was measured by Cell Counting Kit-8,Hoechst33258 staining was used to observe the morphology of apoptotic changes.Mitochondrial membrane potential was detected by JC-1 fluorescent probe,and protein expressions of RXRα,Bcl-2,Bax,caspase-3 and cleaved caspase-3 with Western blotting.RESULTS Compared with control group,the cell viability in model group was decreased(P<0.05).After treated with NCTS in different concentrations,the CCK8 results showed that NCTS in 2μmol·L^-1 had protective effects.Result of Hoechst33258 staining suggested that the apoptosis was notably increased in model group(P<0.05),Meanwhile,the JC-1 results showed that the mitochondrial membrane potential of the model group decreased which was consistent with previous study.impressively,NCTS could restore the mitochondrial membrane potential as well as apoptosis.Fur⁃ther western blot experiments showed that NCTS treat could upregulate Bcl-2 protein,and downregulate the levels of Bax and cleaved caspase-3/caspase-3 ratio.Since RXRαis a critical upstreaming proteins which can directly mediate the apoptosis,we then determined the effect of NCTS on it.Intriguingly,RXRαwas notably activated by NCTS,while the HX531,the antagonist of RXRα,could abolished NCTS'effect when co-treated with NCTS.CONCLUSION NCTS in 2μmol·L^-1 was effective to protect H9C2 cell from H/R-induced cell injury through RXRα-mediated mitochondria apop⁃tosis.Current results provide possible drugs for the treatment of ischemic cardiomyopathy.展开更多
目的使用饮食诱导肥胖(DIO)大鼠模型,探索间歇性缺氧-复氧(IHR)对肥胖大鼠体质量、食水摄入量、循环代谢因子和中枢瘦素注射反应的影响。方法通过12周高脂饮食(HFD)喂养建立DIO大鼠模型,将其随机分为3组并继续HFD喂养:常氧组(NM,n=15)...目的使用饮食诱导肥胖(DIO)大鼠模型,探索间歇性缺氧-复氧(IHR)对肥胖大鼠体质量、食水摄入量、循环代谢因子和中枢瘦素注射反应的影响。方法通过12周高脂饮食(HFD)喂养建立DIO大鼠模型,将其随机分为3组并继续HFD喂养:常氧组(NM,n=15)、间歇性缺氧组(IH:6%O2,30周期/h,8 h/d,4周,n=15),IHR组(缺氧2周后复氧2周,n=15)。记录大鼠体质量、饮食饮水情况,检测循环瘦素、IL-6、Ang-II含量。IHR干预结束后,大鼠接受4μg瘦素侧脑室注射,1 h后处死取材下丘脑及肝脏。通过免疫组化观察下丘脑POMC、FRA-1、FRA-2表达,Western blotting检测下丘脑POMC、pSTAT3、LepR表达,RT-PCR检测下丘脑和肝脏中LepR mRNA含量,对比各组大鼠下丘脑瘦素受体(LepR)及下游通路蛋白的变化。结果IH暴露导致DIO大鼠体质量(P=0.001)和摄食量(P=0.001)增加,全身炎症因子升高(瘦素P=0.004;IL-6 P=0.008;Ang-II P<0.001)。IH抑制下丘脑食欲抑制肽POMC表达(P<0.001 vs NM组),降低反映瘦素反应性神经元活性的FRA-1表达(P<0.001 vs NM组),抑制对瘦素响应的pSTAT3表达(瘦素+vs瘦素-,P=0.241),降低对外源性瘦素给药的反应性(P<0.001 vs NM组),并下调下丘脑和肝脏LepR mRNA含量(P<0.001 vs NM组)。经过2周的复氧治疗后,IH加剧的体质量增加和代谢紊乱能够得到改善,下丘脑瘦素反应性也有所提高。结论IH可能通过下调LepR表达损害下丘脑瘦素信号传导,从而促进肥胖大鼠增重,这可以通过复氧治疗得到改善。展开更多
基金National Natural Science Foundation of China(81822049and 81673712)
文摘OBJECTIVE Neocryptotanshinone(NCTS)is a natural product extracted from traditional Chinese herb Salvia miltiorrhiza Bunge.Previous studies have demonstrated the anti-inflammatory of NCTS in lipopolysaccharide(LPS)-stimulated mouse macrophage(RAW 264.7).However,the protective effect and mechanism of NCTS in cardiomy⁃ocytes are still undefined.This study is to investigate whether NCTS exerts its cardioprotective effect against hypoxia/re⁃oxygenation(H/R)-induced H9C2 cell injury.METHODS The model of H/R injury was established through hypoxia for 8 h and reoxygenation for 12 h in H9C2 cardiomyocytes of rats.Cultured cardiomyocytes were randomly divided into four groups,control group,H/R group,H/R+NCTS pretreated group(1,2,5 and 10μmol·L^-1),and H/R+NCTS+HX531(an RXRαantagonist,2μmol·L^-1)co-treated group.The cell viability was measured by Cell Counting Kit-8,Hoechst33258 staining was used to observe the morphology of apoptotic changes.Mitochondrial membrane potential was detected by JC-1 fluorescent probe,and protein expressions of RXRα,Bcl-2,Bax,caspase-3 and cleaved caspase-3 with Western blotting.RESULTS Compared with control group,the cell viability in model group was decreased(P<0.05).After treated with NCTS in different concentrations,the CCK8 results showed that NCTS in 2μmol·L^-1 had protective effects.Result of Hoechst33258 staining suggested that the apoptosis was notably increased in model group(P<0.05),Meanwhile,the JC-1 results showed that the mitochondrial membrane potential of the model group decreased which was consistent with previous study.impressively,NCTS could restore the mitochondrial membrane potential as well as apoptosis.Fur⁃ther western blot experiments showed that NCTS treat could upregulate Bcl-2 protein,and downregulate the levels of Bax and cleaved caspase-3/caspase-3 ratio.Since RXRαis a critical upstreaming proteins which can directly mediate the apoptosis,we then determined the effect of NCTS on it.Intriguingly,RXRαwas notably activated by NCTS,while the HX531,the antagonist of RXRα,could abolished NCTS'effect when co-treated with NCTS.CONCLUSION NCTS in 2μmol·L^-1 was effective to protect H9C2 cell from H/R-induced cell injury through RXRα-mediated mitochondria apop⁃tosis.Current results provide possible drugs for the treatment of ischemic cardiomyopathy.
文摘目的使用饮食诱导肥胖(DIO)大鼠模型,探索间歇性缺氧-复氧(IHR)对肥胖大鼠体质量、食水摄入量、循环代谢因子和中枢瘦素注射反应的影响。方法通过12周高脂饮食(HFD)喂养建立DIO大鼠模型,将其随机分为3组并继续HFD喂养:常氧组(NM,n=15)、间歇性缺氧组(IH:6%O2,30周期/h,8 h/d,4周,n=15),IHR组(缺氧2周后复氧2周,n=15)。记录大鼠体质量、饮食饮水情况,检测循环瘦素、IL-6、Ang-II含量。IHR干预结束后,大鼠接受4μg瘦素侧脑室注射,1 h后处死取材下丘脑及肝脏。通过免疫组化观察下丘脑POMC、FRA-1、FRA-2表达,Western blotting检测下丘脑POMC、pSTAT3、LepR表达,RT-PCR检测下丘脑和肝脏中LepR mRNA含量,对比各组大鼠下丘脑瘦素受体(LepR)及下游通路蛋白的变化。结果IH暴露导致DIO大鼠体质量(P=0.001)和摄食量(P=0.001)增加,全身炎症因子升高(瘦素P=0.004;IL-6 P=0.008;Ang-II P<0.001)。IH抑制下丘脑食欲抑制肽POMC表达(P<0.001 vs NM组),降低反映瘦素反应性神经元活性的FRA-1表达(P<0.001 vs NM组),抑制对瘦素响应的pSTAT3表达(瘦素+vs瘦素-,P=0.241),降低对外源性瘦素给药的反应性(P<0.001 vs NM组),并下调下丘脑和肝脏LepR mRNA含量(P<0.001 vs NM组)。经过2周的复氧治疗后,IH加剧的体质量增加和代谢紊乱能够得到改善,下丘脑瘦素反应性也有所提高。结论IH可能通过下调LepR表达损害下丘脑瘦素信号传导,从而促进肥胖大鼠增重,这可以通过复氧治疗得到改善。
