丙泊酚对氯化钴诱导人心肌AC16细胞低氧损伤的保护作用
|
摘要
目的:研究丙泊酚预处理对氯化钴(cobalt chloride,CoCl2)诱导离体人心肌AC16细胞低氧损伤的影响及其相关的分子机制。方法:以CoCl2处理的人心肌AC16细胞作为心肌细胞低氧的体外模型。将AC16细胞分为对照组、CoCl2诱导低氧组(CoCl2组)和丙泊酚+CoCl2诱导低氧组(Propofol+CoCl2组)。采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)评估细胞活力;采用流式细胞术分析AC16细胞凋亡比率(apoptosis ratio,AR)和线粒体膜电位(mitochondrial membrane potential,Δψm);采用对线粒体活性氧(reactive oxygen species,ROS)敏感的荧光探针测定各组AC16细胞中ROS含量,并检测AC16细胞中的丙二醛(malondialdehyde,MDA)和超氧化物歧化酶(superoxide dismutase,SOD)水平;采用Western印迹评估c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)和p38信号转导途径的激活。结果:1)与对照组相比,以500μmol/L CoCl2处理12 h的CoCl2组AC16细胞活力显著降低(P<0.01);2)与对照组相比,CoCl2组和Propofol+CoCl2组AC16细胞的AR均显著升高,而Δψm均显著降低(均P<0.01);与CoCl2组相比,Propofol+CoCl2组AC16细胞的AR显著降低,而Δψm显著升高(均P<0.05);3)与对照组相比,CoCl2组的ROS和MDA水平显著升高,SOD水平显著降低(均P<0.01);与对照组相比,Propofol+CoCl2组ROS和MDA水平显著升高,SOD水平显著降低(均P<0.05);4)与对照组相比,CoCl2组JNK和p38的磷酸化水平显著升高(均P<0.05);与CoCl2组相比,Propofol+CoCl2组JNK和p38的磷酸化水平显著降低(均P<0.05)。结论:丙泊酚预处理可能通过抑制JNK和p38信号通路的激活,从而保护人心肌AC16细胞免受CoCl2诱导的低氧损伤。
Objective: To explore the effect of propofol on human cardiac AC16 cells under CoCl_2-induced hypoxic injury and the possible mechanisms.Methods: Human AC16 cardiomyocytes were treated with cobalt chloride(CoCl_2) to mimic hypoxic condition in cultured cardiomyocytes. The AC16 cells were divided into 3 groups: a control group, a CoCl_2 hypoxia group(CoCl_2 group), and a propofol+CoCl_2 group(propofol+CoCl_2 group). The cell viability was assessed by cell counting kit-8(CCK-8). Cell apoptosis ratio(AR) and the mitochondrial membrane potential(Δψm) were detected by flow cytometry. The reactive oxygen species(ROS) production in AC16 cells were determined with the ROS-sensitive fluorescent probe. Meanwhile, total intracellular levels of malondialdehyde(MDA) and superoxide dismutase(SOD) in AC16 cells were detected with commercially available kits. Western blot was used to evaluate the activation of c-Jun N-terminal kinase(JNK) and p38 signaling pathways.Results: 1) Compared with the control group, AC16 cell viability was decreased significantly in the CoCl_2 group following the treatment with 500 μmol/L CoCl_2(P<0.01); 2) Compared with the control group, AR value in AC16 cells was increased significantly in the CoCl_2 group, while Δψm was decreased significantly(all P<0.01). Compared with the CoCl_2 group, AR value in AC16 cells was decreased significantly in the propofol+CoCl_2 group, while Δψm was increased significantly(both P<0.05); 3) Compared with the control group, the levels of ROS and MDA were increased significantly, and the level of SOD was significantly decreased in the CoCl_2 group(all P<0.01).Compared with the CoCl_2 group, the ROS and MDA levels in the propofol+CoCl_2 group were increased significantly and the SOD levels were decreased significantly(all P<0.05); 4) Compared with the control group, the phosphorylation levels of JNK and p38 were increased significantly(both P<0.05) in the CoCl_2 group. Compared with the CoCl_2 group, the phosphorylation levels of JNK and p38 were decreased significantly in the propofol+CoCl_2 group(both P<0.05).Conclusion: The pretreatment with propofol may protect human cardiac AC16 cells from the chemical hypoxia-induced injury through regulation of JNK and p38 signaling pathways.
Objective: To explore the effect of propofol on human cardiac AC16 cells under CoCl_2-induced hypoxic injury and the possible mechanisms.Methods: Human AC16 cardiomyocytes were treated with cobalt chloride(CoCl_2) to mimic hypoxic condition in cultured cardiomyocytes. The AC16 cells were divided into 3 groups: a control group, a CoCl_2 hypoxia group(CoCl_2 group), and a propofol+CoCl_2 group(propofol+CoCl_2 group). The cell viability was assessed by cell counting kit-8(CCK-8). Cell apoptosis ratio(AR) and the mitochondrial membrane potential(Δψm) were detected by flow cytometry. The reactive oxygen species(ROS) production in AC16 cells were determined with the ROS-sensitive fluorescent probe. Meanwhile, total intracellular levels of malondialdehyde(MDA) and superoxide dismutase(SOD) in AC16 cells were detected with commercially available kits. Western blot was used to evaluate the activation of c-Jun N-terminal kinase(JNK) and p38 signaling pathways.Results: 1) Compared with the control group, AC16 cell viability was decreased significantly in the CoCl_2 group following the treatment with 500 μmol/L CoCl_2(P<0.01); 2) Compared with the control group, AR value in AC16 cells was increased significantly in the CoCl_2 group, while Δψm was decreased significantly(all P<0.01). Compared with the CoCl_2 group, AR value in AC16 cells was decreased significantly in the propofol+CoCl_2 group, while Δψm was increased significantly(both P<0.05); 3) Compared with the control group, the levels of ROS and MDA were increased significantly, and the level of SOD was significantly decreased in the CoCl_2 group(all P<0.01).Compared with the CoCl_2 group, the ROS and MDA levels in the propofol+CoCl_2 group were increased significantly and the SOD levels were decreased significantly(all P<0.05); 4) Compared with the control group, the phosphorylation levels of JNK and p38 were increased significantly(both P<0.05) in the CoCl_2 group. Compared with the CoCl_2 group, the phosphorylation levels of JNK and p38 were decreased significantly in the propofol+CoCl_2 group(both P<0.05).Conclusion: The pretreatment with propofol may protect human cardiac AC16 cells from the chemical hypoxia-induced injury through regulation of JNK and p38 signaling pathways.
引文
[1]Gray LD,Morris C.The principles and conduct of anaesthesia for emergency surgery[J].Anaesthesia,2013,68(Suppl 1):14-29.
[2]Abubaih A,Weissman C.Anesthesia for patients with concomitant sepsis and cardiac dysfunction[J].Anesthesiol Clin,2016,34(4):761-774.
[3]孔令恒,梁飞,陈玉龙,等.钠钙交换体激活CaMK II介导大鼠离体心脏缺血再灌注损伤[J].中南大学学报(医学版),2018,43(1):28-34.KONG Lingheng,LIANG Fei,CHEN Yulong,et al.Na+/Ca2+exchanger mediates ischemia-reperfusion injury by activation of CaMKII in isolated rat heart[J].Journal of Central South University.Medicine Science,2018,43(1):28-34.
[4]Tsai YC,Huang CC,Chu LM,et al.Differential influence of propofol on different cell types in terms of the expression of various oxidative stress-related enzymes in an experimental endotoxemia model[J].Acta Anaesthesiol Taiwan,2012,50(4):159-166.
[5]Wang B,Luo T,Chen D,et al.Propofol reduces apoptosis and upregulates endothelial nitric oxide synthase protein expression in hydrogen peroxide-stimulated human umbilical vein endothelial cells[J].Anesth Analg,2007,105(4):1027-1033.
[6]Zhang J,Xia Y,Xu Z,et al.Propofol suppressed hypoxia/reoxygenation-induced apoptosis in HBVSMC by regulation of the expression of Bcl-2,Bax,caspase3,Kir6.1,and p-JNK[J].Oxid Med Cell Longev,2016,2016:1518738.
[7]Lu Y,Gu Y,Ding X,et al.Intracellular Ca2+homeostasis and JAK1/STAT3 pathway are involved in the protective effect of propofol on BV2 microglia against hypoxia-induced inflammation and apoptosis[J].PLoS One,2017,12(5):e0178098.
[8]Yu X,Sun X,Zhao M,et al.Propofol attenuates myocardial ischemia reperfusion injury partly through inhibition of resident cardiac mast cell activation[J].Int Immunopharmacol,2018,54(2):267-274.
[9]Mao SY,Meng XY,Xu ZW,et al.The role of ZFP580,a novel zinc finger protein,in TGF-mediated cytoprotection against chemical hypoxia induced apoptosis in H9c2 cardiac myocytes[J].Mol Med Rep,2017,15(4):2154-2162.
[10]Jeon YJ,Kim HS,Song KS,et al.Protective effect of dieckol against chemical hypoxia-induced cytotoxicity in primary cultured mouse hepatocytes[J].Drug Chem Toxicol,2015,38(2):180-187.
[11]Feng AY,Kaye AD,Kaye RJ,et al.Novel propofol derivatives and implications for anesthesia practice[J].J Anaesthesiol Clin Pharmacol,2017,33(1):9-15.
[12]Liu XR,Cao L,Li T,et al.Propofol attenuates H2O2-induced oxidative stress and apoptosis via the mitochondria-and ERmedicated pathways in neonatal rat cardiomyocytes[J].Apoptosis,2017,22(5):639-646.
[13]Chen XH,Zhou X,Yang XY,et al.Propofol protects against H2O2-induced oxidative injury in differentiated PC12 cells via inhibition of Ca2+-dependent NADPH oxidase[J].Cell Mol Neurobiol,2016,36(4):541-551.
[14]Wang Z,Yang P,Qi Y.Role of microRNA-134 in the neuroprotective effects of propofol against oxygen-glucosedeprivation and related mechanisms[J].Int J Clin Exp Med,2015,8(11):20617-20623.
[15]Gokcinar D,Ergin V,Cumaoglu A,et al.Effects of ketamine,propofol,and ketofol on proinflammatory cytokines and markers of oxidative stress in a rat model of endotoxemia-induced acute lung injury[J].Acta Biochim Pol,2013,60(3):451-456.
[16]Jiang Z,Song F,Li Y,et al.Capsular polysaccharide of mycoplasma ovipneumoniae induces sheep airway epithelial cell apoptosis via ROS-dependent JNK/P38 MAPK pathways[J].Oxid Med Cell Longev,2017,2017:6175841.
[17]Xiong T,Dong W,Fu H,et al.Involvement of the nuclear factor-κB pathway in the adhesion of neutrophils to renal tubular cells after injury induced by neonatal postasphyxial serum[J].Mol Cell Biochem,2014,388(1/2):85-94.
[2]Abubaih A,Weissman C.Anesthesia for patients with concomitant sepsis and cardiac dysfunction[J].Anesthesiol Clin,2016,34(4):761-774.
[3]孔令恒,梁飞,陈玉龙,等.钠钙交换体激活CaMK II介导大鼠离体心脏缺血再灌注损伤[J].中南大学学报(医学版),2018,43(1):28-34.KONG Lingheng,LIANG Fei,CHEN Yulong,et al.Na+/Ca2+exchanger mediates ischemia-reperfusion injury by activation of CaMKII in isolated rat heart[J].Journal of Central South University.Medicine Science,2018,43(1):28-34.
[4]Tsai YC,Huang CC,Chu LM,et al.Differential influence of propofol on different cell types in terms of the expression of various oxidative stress-related enzymes in an experimental endotoxemia model[J].Acta Anaesthesiol Taiwan,2012,50(4):159-166.
[5]Wang B,Luo T,Chen D,et al.Propofol reduces apoptosis and upregulates endothelial nitric oxide synthase protein expression in hydrogen peroxide-stimulated human umbilical vein endothelial cells[J].Anesth Analg,2007,105(4):1027-1033.
[6]Zhang J,Xia Y,Xu Z,et al.Propofol suppressed hypoxia/reoxygenation-induced apoptosis in HBVSMC by regulation of the expression of Bcl-2,Bax,caspase3,Kir6.1,and p-JNK[J].Oxid Med Cell Longev,2016,2016:1518738.
[7]Lu Y,Gu Y,Ding X,et al.Intracellular Ca2+homeostasis and JAK1/STAT3 pathway are involved in the protective effect of propofol on BV2 microglia against hypoxia-induced inflammation and apoptosis[J].PLoS One,2017,12(5):e0178098.
[8]Yu X,Sun X,Zhao M,et al.Propofol attenuates myocardial ischemia reperfusion injury partly through inhibition of resident cardiac mast cell activation[J].Int Immunopharmacol,2018,54(2):267-274.
[9]Mao SY,Meng XY,Xu ZW,et al.The role of ZFP580,a novel zinc finger protein,in TGF-mediated cytoprotection against chemical hypoxia induced apoptosis in H9c2 cardiac myocytes[J].Mol Med Rep,2017,15(4):2154-2162.
[10]Jeon YJ,Kim HS,Song KS,et al.Protective effect of dieckol against chemical hypoxia-induced cytotoxicity in primary cultured mouse hepatocytes[J].Drug Chem Toxicol,2015,38(2):180-187.
[11]Feng AY,Kaye AD,Kaye RJ,et al.Novel propofol derivatives and implications for anesthesia practice[J].J Anaesthesiol Clin Pharmacol,2017,33(1):9-15.
[12]Liu XR,Cao L,Li T,et al.Propofol attenuates H2O2-induced oxidative stress and apoptosis via the mitochondria-and ERmedicated pathways in neonatal rat cardiomyocytes[J].Apoptosis,2017,22(5):639-646.
[13]Chen XH,Zhou X,Yang XY,et al.Propofol protects against H2O2-induced oxidative injury in differentiated PC12 cells via inhibition of Ca2+-dependent NADPH oxidase[J].Cell Mol Neurobiol,2016,36(4):541-551.
[14]Wang Z,Yang P,Qi Y.Role of microRNA-134 in the neuroprotective effects of propofol against oxygen-glucosedeprivation and related mechanisms[J].Int J Clin Exp Med,2015,8(11):20617-20623.
[15]Gokcinar D,Ergin V,Cumaoglu A,et al.Effects of ketamine,propofol,and ketofol on proinflammatory cytokines and markers of oxidative stress in a rat model of endotoxemia-induced acute lung injury[J].Acta Biochim Pol,2013,60(3):451-456.
[16]Jiang Z,Song F,Li Y,et al.Capsular polysaccharide of mycoplasma ovipneumoniae induces sheep airway epithelial cell apoptosis via ROS-dependent JNK/P38 MAPK pathways[J].Oxid Med Cell Longev,2017,2017:6175841.
[17]Xiong T,Dong W,Fu H,et al.Involvement of the nuclear factor-κB pathway in the adhesion of neutrophils to renal tubular cells after injury induced by neonatal postasphyxial serum[J].Mol Cell Biochem,2014,388(1/2):85-94.