小檗碱对波动性高糖诱导的人冠状动脉内皮细胞凋亡的影响
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摘要
目的研究小檗碱对波动性高糖所致人冠状动脉内皮细胞(HCAECs)凋亡的影响。方法分离、鉴定并培养HCAECs,将HCAECs分为5组:正常葡萄糖组(5.5 mmol/L葡萄糖)、持续性高糖组(25 mmol/L葡萄糖)、波动性高糖组(5.5、25 mmol/L葡萄糖每24小时波动1次)、波动性高渗环境(不加入葡萄糖,加入与波动性高糖组相同浓度的甘露醇,保持与波动性高糖组相同的波动性高渗环境)、波动性高糖+小檗碱干预组(5.5、25mmol/L葡萄糖+50μmol/L小檗碱每24小时波动1次),各组细胞每24小时换液1次,共培养7 d后进行后期实验。检测各组HCAECs的细胞活力及凋亡情况,利用实时荧光定量(q RT-PCR)及Western blotting技术检测细胞凋亡相关蛋白Caspase-3表达量。结果与持续性高糖组相比,波动性高糖组HCAECs凋亡更显著(P<0.05),波动性高糖组Caspase-3蛋白及m RNA表达水平明显增加(P<0.05);但波动性高糖+小檗碱干预组较波动性高糖组HCAECs凋亡明显减弱(P<0.05),小檗碱可以明显下调波动性高糖诱导的Caspase-3表达(P<0.05)。结论波动性高糖较持续性高糖更易降低HCAECs活力,促进HCAECs凋亡,但小檗碱显著减弱波动性高糖环境下HCAECs凋亡,对HCAECs具有明显的保护作用。
Objective To investigate the effect of berberine on the apoptosis of human coronary artery endothelial cells(HCAECs)induced by intermittent high glucose in vitro. Methods HCAECs were isolated, identified, cultured, and divided into five groups:Group A was the normal glucose group(5.5 mmol/L glucose, NG); Group B was the persistent high glucose group(25 mmol/L glucose,PHG); Group C was the intermittent high glucose group(5.5 mmol/L and 25 mmol/L glucose fluctuated every 24 h, IHG); Group D was in a volatile hyperosmotic environment(5.5 mmol/L and 25 mmol/L mannitol alternating every 24 h to maintain the same osmotic pressure as IHG, OC); Group E was the fluctuation of hyperglycemia + berberine intervention group(5.5 mmol/L and 25 mmol/L glucose + 50 μmol/L berberine fluctuated every 24 h, IHG + BBR). The cells of each group were changed every 24 h, and the culture was stopped after 7 d, the cell viability and apoptosis were detected in each group. The expression of the apoptosis related protein Caspase-3 was determined by qRT-PCR and Western blotting. Results Compared with the persistent high glucose group, the apoptosis of HCAECs in the intermittent high glucose group was more significant(P < 0.05), but the apoptosis of HCAECs in the fluctuation of hyperglycemia + berberine intervention group was more significantly reduced than that in the intermittent high glucose group(P < 0.05). Berberine significantly reduced the expression of Caspase-3 induced by intermittent high glucose(P < 0.05)Conclusion Intermittent high glucose decreased the activity of HCAECs more easily than the persistent high glucose,and promoted the apoptosis of HCAECs. But berberine significantly reduced the apoptosis of HCAECs under intermittent high glucose, which had a significant protective effect on HCAECs.
Objective To investigate the effect of berberine on the apoptosis of human coronary artery endothelial cells(HCAECs)induced by intermittent high glucose in vitro. Methods HCAECs were isolated, identified, cultured, and divided into five groups:Group A was the normal glucose group(5.5 mmol/L glucose, NG); Group B was the persistent high glucose group(25 mmol/L glucose,PHG); Group C was the intermittent high glucose group(5.5 mmol/L and 25 mmol/L glucose fluctuated every 24 h, IHG); Group D was in a volatile hyperosmotic environment(5.5 mmol/L and 25 mmol/L mannitol alternating every 24 h to maintain the same osmotic pressure as IHG, OC); Group E was the fluctuation of hyperglycemia + berberine intervention group(5.5 mmol/L and 25 mmol/L glucose + 50 μmol/L berberine fluctuated every 24 h, IHG + BBR). The cells of each group were changed every 24 h, and the culture was stopped after 7 d, the cell viability and apoptosis were detected in each group. The expression of the apoptosis related protein Caspase-3 was determined by qRT-PCR and Western blotting. Results Compared with the persistent high glucose group, the apoptosis of HCAECs in the intermittent high glucose group was more significant(P < 0.05), but the apoptosis of HCAECs in the fluctuation of hyperglycemia + berberine intervention group was more significantly reduced than that in the intermittent high glucose group(P < 0.05). Berberine significantly reduced the expression of Caspase-3 induced by intermittent high glucose(P < 0.05)Conclusion Intermittent high glucose decreased the activity of HCAECs more easily than the persistent high glucose,and promoted the apoptosis of HCAECs. But berberine significantly reduced the apoptosis of HCAECs under intermittent high glucose, which had a significant protective effect on HCAECs.
引文
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[18]Zhou H,Sun Y,Zhang L,et al.The RhoA/ROCK pathway mediates high glucose-induced cardiomyocyte apoptosis via oxidative stress,JNK,and p38MAPK pathways[J].Diabetes Metab Res Rev,2018,34(6):3022-3028.
[19]Nazanin N S,Saeedeh S F,Reza F,et al.Phosphodiesterase 4 and 7 inhibitors produce protective effects against high glucose-induced neurotoxicity in PC12 cells via modulation of the oxidative stress,apoptosis and inflammation pathways[J].Metab Brain Dis,2018,33(4):1293-1306.
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[22]Xue M,Qiqige C,Zhang Q,et al.Effects of tumor necrosis factor alpha(TNF-alpha)and interleukina 10(IL-10)on intercellular cell adhesion molecule-1(ICAM-1)and cluster of differentiation 31(CD31)in human coronary artery endothelial cells[J].Med Sci Monit,2018,6(27):4433-4439.
[23]Shimoda Y,Tagaya Y,Saito T,et al.Continuous or transient high level of glucose exposure differentially increases coronary artery endothelial cell proliferation and human colon cancer cell proliferation[J].Cell J,2017,19(Suppl 1):106-112.
[24]Song C L,Liu B,Shi Y F,et al.Micro RNA-130a alleviates human coronary artery endothelial cell injury and inflammatory responses by targeting PTEN via activating PI3K/Akt/eNOS signaling pathway[J].Oncotarget,2016,7(44):71922-71936.
[25]Cimen T,Efe T H,Akyel A,et al.Human endothelial cell-specific molecule-1(endocan)and coronary artery disease and microvascular angina[J].Angiology,2016,67(9):846-853.
[26]Zhu Y,Yi Y,Bai B,et al.The silencing of replication protein A1 induced cell apoptosis via regulating Caspase3[J].Life Sci,2018,5(15):141-149.
[27]Hou L,Zhang Y,Yu B,et al.Oocyte-G1 promotes male germ cell apoptosis through activation of Caspase-3[J].Gene,2018,9(5):22-30.
[28]宋兆峰,杜波,季晓平.多二磷酸腺苷-核糖聚合酶在大鼠心肌缺血再灌注损伤中的作用及其对核转录因子κB活性和炎症因子表达的调节[J].中华心血管病杂志,2012,40(12):997-1002.
[29]李向东,杨跃进,赵京林,等.缺血预适应改善猪急性心肌梗死再灌注后无复流和再灌注损伤的作用与通过蛋白激酶A通路减轻心肌水肿有关[J].中华心血管病杂志,2012,40(11):945-951.
[30]李美平,孙国杰,尤行宏.头针对心肌缺血再灌注大鼠心交感放电、心肌β1-肾上腺素受体蛋白表达及血浆去甲肾上腺素含量的影响[J].针刺研究,2012,37(5):385-389.
[31]Yu L,Li F,Zhao G,et al.Protective effect of berberine against myocardial ischemia reperfusion injury:Role of Notch1/Hes1-PTEN/Akt signaling[J].Apoptosis,2015,20(6):796-810.
[32]Wang Y,Liu J,Ma A,et al.Cardioprotective effect of berberine against myocardial ischemia/reperfusion injury via attenuating mitochondrial dysfunction and apoptosis[J].Int J Clin Exp Med,2015,8(8):14513-14519.
[33]姜甜,贾友宏,李一石.小檗碱调脂作用机制的研究进展[J].现代药物与临床,2016,31(5):727-730.
[34]沈宁,李彩娜,环奕,等.小檗碱调节血糖血脂代谢紊乱机制研究进展[J].药学学报,2010,45(6):699-704.
[2]Lunder M,Janic M,Sabovic M.Prevention of vascular complications in diabetes mellitus patients:Focus on the arterial wall[J].Curr Vasc Pharmacol,2018,17(1):6-15.
[3]Li H,Luo H Y,Liu Q,et al.Intermittent high glucose exacerbates A-FABP activation and inflammatory response through TLR4-JNK signaling in THP-1 cells[J].J Immunol Res,2018,4(11):272-281.
[4]Zhang W,Song J,Zhang Y,et al.Intermittent high glucose-induced oxidative stress modulates retinal pigmented epithelial cell autophagy and promotes cell survival via increased HMGB1[J].BMC Ophthalmol,2018,18(1):192-200.
[5]Tao S,Ren Y,Zheng H,et al.Salvianolic acid B inhibits intermittent high glucose-induced INS-1 cell apoptosis through regulation of Bcl-2 proteins and mitochondrial membrane potential[J].Eur J Pharmacol,2017,11(5):56-62.
[6]Kim Y J,Park S M,Jung H S,et al.Ginsenoside Rg3prevents INS-1 cell death from intermittent high glucose stress[J].Islets,2016,8(3):57-64.
[7]Wang L,Ma H,Xue Y,et al.Berberine inhibits the ischemia-reperfusion injury induced inflammatory response and apoptosis of myocardial cells through the phosphoinositide 3-kinase/RAC-alpha serine/threonineprotein kinase and nuclear factor-kappaB signaling pathways[J].Exp Ther Med,2018,15(2):1225-1232.
[8]Zhao L,Cang Z,Sun H,et al.Berberine improves glucogenesis and lipid metabolism in nonalcoholic fatty liver disease[J].BMC Endocr Disord,2017,17(1):13-21.
[9]Yao Q,Xu D,Li H L,et al.Berberine promoted myocardial protection of postoperative patients through regulating myocardial autophagy[J].Biomed Pharmacother,2018,9(15):1050-1053.
[10]Zhu Q W,Li Y G.Berberine attenuates myocardial ischemia reperfusion injury by suppressing the activation of PI3K/AKT signaling[J].Exp Ther Med,2016,11(3):978-984.
[11]王灿,蒋建东,孔维佳.小檗碱治疗代谢性疾病抗炎作用的研究进展[J].药物评价研究,2018,41(4):692-696.
[12]Zhao G L,Yu L M,Gao W L,et al.Berberine protects rat heart from ischemia/reperfusion injury via activating JAK2/STAT3 signaling and attenuating endoplasmic reticulum stress[J].Acta Pharmacol Sin,2016,37:354-367.
[13]Zhang Z,Li J,Yang L,et al.The Cytotoxic role of intermittent high glucose on apoptosis and cell viability in pancreatic beta cells[J].J Diabetes Res,2014,doi:org/10.1155/2014/712781.
[14]Zhong Y,Wang J J,Zhang S X.Intermittent but not constant high glucose induces ER stress and inflammation in human retinal pericytes[J].Adv Exp Med Biol,2012,7(23):285-292.
[15]Sun L Q,Chen Y Y,Wang X,et al.The protective effect of alpha lipoic acid on Schwann cells exposed to constant or intermittent high glucose[J].Biochem Pharmacol,2012,84(7):961-973.
[16]Maeda M,Hayashi T,Mizuno N,et al.Intermittent high glucose implements stress-induced senescence in human vascular endothelial cells:Role of superoxide production by NADPH oxidase[J].PLoS One,2015,10(4):e0123169.
[17]Wei J,Wu H,Zhang H,et al.Anthocyanins inhibit high glucose-induced renal tubular cell apoptosis caused by oxidative stress in db/db mice[J].Int J Mol Med,2018,41(3):1608-1618.
[18]Zhou H,Sun Y,Zhang L,et al.The RhoA/ROCK pathway mediates high glucose-induced cardiomyocyte apoptosis via oxidative stress,JNK,and p38MAPK pathways[J].Diabetes Metab Res Rev,2018,34(6):3022-3028.
[19]Nazanin N S,Saeedeh S F,Reza F,et al.Phosphodiesterase 4 and 7 inhibitors produce protective effects against high glucose-induced neurotoxicity in PC12 cells via modulation of the oxidative stress,apoptosis and inflammation pathways[J].Metab Brain Dis,2018,33(4):1293-1306.
[20]Jadaun P,Yadav D,Bisen P S.Spirulina platensis prevents high glucose-induced oxidative stress mitochondrial damage mediated apoptosis in cardiomyoblasts[J].Cytotechnology,2018,70(2):523-536.
[21]Fei Y,Sun L,Yuan C,et al.CFTR ameliorates high glucose-induced oxidative stress and inflammation by mediating the NF-kappaB and MAPK signaling pathways in endothelial cells[J].Int J Mol Med,2018,41(6):3501-3508.
[22]Xue M,Qiqige C,Zhang Q,et al.Effects of tumor necrosis factor alpha(TNF-alpha)and interleukina 10(IL-10)on intercellular cell adhesion molecule-1(ICAM-1)and cluster of differentiation 31(CD31)in human coronary artery endothelial cells[J].Med Sci Monit,2018,6(27):4433-4439.
[23]Shimoda Y,Tagaya Y,Saito T,et al.Continuous or transient high level of glucose exposure differentially increases coronary artery endothelial cell proliferation and human colon cancer cell proliferation[J].Cell J,2017,19(Suppl 1):106-112.
[24]Song C L,Liu B,Shi Y F,et al.Micro RNA-130a alleviates human coronary artery endothelial cell injury and inflammatory responses by targeting PTEN via activating PI3K/Akt/eNOS signaling pathway[J].Oncotarget,2016,7(44):71922-71936.
[25]Cimen T,Efe T H,Akyel A,et al.Human endothelial cell-specific molecule-1(endocan)and coronary artery disease and microvascular angina[J].Angiology,2016,67(9):846-853.
[26]Zhu Y,Yi Y,Bai B,et al.The silencing of replication protein A1 induced cell apoptosis via regulating Caspase3[J].Life Sci,2018,5(15):141-149.
[27]Hou L,Zhang Y,Yu B,et al.Oocyte-G1 promotes male germ cell apoptosis through activation of Caspase-3[J].Gene,2018,9(5):22-30.
[28]宋兆峰,杜波,季晓平.多二磷酸腺苷-核糖聚合酶在大鼠心肌缺血再灌注损伤中的作用及其对核转录因子κB活性和炎症因子表达的调节[J].中华心血管病杂志,2012,40(12):997-1002.
[29]李向东,杨跃进,赵京林,等.缺血预适应改善猪急性心肌梗死再灌注后无复流和再灌注损伤的作用与通过蛋白激酶A通路减轻心肌水肿有关[J].中华心血管病杂志,2012,40(11):945-951.
[30]李美平,孙国杰,尤行宏.头针对心肌缺血再灌注大鼠心交感放电、心肌β1-肾上腺素受体蛋白表达及血浆去甲肾上腺素含量的影响[J].针刺研究,2012,37(5):385-389.
[31]Yu L,Li F,Zhao G,et al.Protective effect of berberine against myocardial ischemia reperfusion injury:Role of Notch1/Hes1-PTEN/Akt signaling[J].Apoptosis,2015,20(6):796-810.
[32]Wang Y,Liu J,Ma A,et al.Cardioprotective effect of berberine against myocardial ischemia/reperfusion injury via attenuating mitochondrial dysfunction and apoptosis[J].Int J Clin Exp Med,2015,8(8):14513-14519.
[33]姜甜,贾友宏,李一石.小檗碱调脂作用机制的研究进展[J].现代药物与临床,2016,31(5):727-730.
[34]沈宁,李彩娜,环奕,等.小檗碱调节血糖血脂代谢紊乱机制研究进展[J].药学学报,2010,45(6):699-704.
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