干扰S100钙结合蛋白B表达对多柔比星诱导心肌细胞凋亡的影响及相关机制
|
摘要
目的探讨干扰S100钙结合蛋白B(S100B)表达对多柔比星诱导心肌细胞凋亡的影响及其相关机制。方法分离培养SD大鼠心肌细胞,共分4组:(1)空白对照组:正常培养,不处理;(2)多柔比星组:2μmol/L多柔比星处理2 h;(3)阴性对照组:2μmol/L多柔比星处理2 h+转染S100B小干扰RNA(siRNA)阴性对照;(4) S100B组:2μmol/L多柔比星处理2 h+转染S100B siRNA。采用噻唑蓝比色法检测各组心肌细胞存活率,采用TUNEL法检测各组心肌细胞凋亡率,采用Elisa法检测各组心肌细胞上清液中肿瘤坏死因子(TNF)-α、白介素(IL)-1β、IL-6水平,采用蛋白印记法检测各组心肌细胞S100B、Caspase-3、Bax、B淋巴细胞瘤(Bcl)-2表达。结果分离培养的大鼠心肌细胞状态良好,阳性率可达(92.33±5.16)%。与空白对照组比较,多柔比星组心肌细胞存活率和Bcl-2蛋白表达降低;心肌细胞凋亡率,TNF-α、IL-1β、IL-6水平,S100B、Caspase-3、Bax蛋白表达增加(P <0.01)。与多柔比星组比较,S100B组心肌细胞存活率和Bcl-2蛋白表达增加;心肌细胞凋亡率,TNF-α、IL-1β、IL-6水平,S100B、Caspase-3、Bax蛋白表达降低(P <0.01)。结论干扰S100B表达能够抑制多柔比星诱导的心肌细胞凋亡,其机制可能与抑制炎性损伤和调节凋亡相关蛋白有关。
Objective To investigate the effect of interfering S100 calcium-binding protein B(S100 B) expression on the doxorubicin induced cardiomyocyte apoptosis and its related mechanisms.Methods The cardiac myocytes cells were isolated and cultured from SD rats.The experiment was divided into 4 groups:(1)blank control group:without treated;(2)doxorubicin group:treated with doxorubicin(2 μmol/L) for 2 h;(3)negative control group:treated with doxorubicin(2 μmol/L) for 2 h,and transfected with S100 B small interference RNA(siRNA) negative control;(4)S100 B group:treated with doxorubicin(2 μmol/L) for 2 h,and transfected with S100 B siRNA.The survival rate of cardiomyocyte was detected by thiazolium colorimetry,the apoptosis rate of cardiomyocyte was detected by TUNEL,the levels of TNF-α,IL-1β and IL-6 in the supernatant were detected by Elisa method,and the expressions of S100 B,Caspase-3,Bax and Bcl-2 were detected by Western blot.Results The purity of cardiomyocyte reached(92.33±5.16)%.Compared with the blank control group,the survival rate and Bcl-2 expression of cardiomyocyte in doxorubicin group were decreased,the apoptotic rate of cardiomyocyte,the levels of TNF-α,IL-1β and IL-6,the expressions of S100 B,Caspase-3 and Bax were increased(P<0.01).Compared with the doxorubicin group,the survival rate of cardiomyocyte and the expression of Bcl-2 protein in S100 B group were increased.Apoptotic rate,TNF-α,IL-1β andIL-6,S100 B,Caspase-3,Bax expression were decreased(P<0.01).Conclusion Interfering S100 B expression can inhibit doxorubicin induced cardiomyocyte apoptosis,and its mechanism may be related to inhibiting inflammatory injury and regulating apoptosis related proteins.
Objective To investigate the effect of interfering S100 calcium-binding protein B(S100 B) expression on the doxorubicin induced cardiomyocyte apoptosis and its related mechanisms.Methods The cardiac myocytes cells were isolated and cultured from SD rats.The experiment was divided into 4 groups:(1)blank control group:without treated;(2)doxorubicin group:treated with doxorubicin(2 μmol/L) for 2 h;(3)negative control group:treated with doxorubicin(2 μmol/L) for 2 h,and transfected with S100 B small interference RNA(siRNA) negative control;(4)S100 B group:treated with doxorubicin(2 μmol/L) for 2 h,and transfected with S100 B siRNA.The survival rate of cardiomyocyte was detected by thiazolium colorimetry,the apoptosis rate of cardiomyocyte was detected by TUNEL,the levels of TNF-α,IL-1β and IL-6 in the supernatant were detected by Elisa method,and the expressions of S100 B,Caspase-3,Bax and Bcl-2 were detected by Western blot.Results The purity of cardiomyocyte reached(92.33±5.16)%.Compared with the blank control group,the survival rate and Bcl-2 expression of cardiomyocyte in doxorubicin group were decreased,the apoptotic rate of cardiomyocyte,the levels of TNF-α,IL-1β and IL-6,the expressions of S100 B,Caspase-3 and Bax were increased(P<0.01).Compared with the doxorubicin group,the survival rate of cardiomyocyte and the expression of Bcl-2 protein in S100 B group were increased.Apoptotic rate,TNF-α,IL-1β andIL-6,S100 B,Caspase-3,Bax expression were decreased(P<0.01).Conclusion Interfering S100 B expression can inhibit doxorubicin induced cardiomyocyte apoptosis,and its mechanism may be related to inhibiting inflammatory injury and regulating apoptosis related proteins.
引文
[1]武旖旎,李东丽,余小燕,等.PI3K/Akt通路介导自噬在七氟烷后处理阿霉素心肌细胞损伤中的研究[J].中华全科医学,2017(1):17-19.
[2]黄俊辉,张曦蓓,肖佳,等.米非司酮逆转人乳腺癌细胞MCF-7/ADR耐多柔比星机制的探讨[J].实用肿瘤杂志,2011,26(6):584-588.
[3]杜天幸,杨继元.蒽环类抗肿瘤药物对心脏毒性防治的研究进展[J].华南国防医学杂志,2015,29(1):80-82.
[4]马军,沈志祥,秦叔逵.防治蒽环类抗肿瘤药物心脏毒性的中国专家共识(2011版)[J].临床肿瘤学杂志,2011,16(12):11 22-11 29.
[5]Tsoporis J N,Marks A,Haddad A,et al.S100B expression modulates left ventricular remodeling after myocardial infarction in mice[J].Circulation,2005,111(5):598-606.
[6]Yan L,Mathew L,Chellan B,et al.S100/calgranulin-mediated inflammation accelerates left ventricular hypertrophy and aortic valve sclerosis in chronic kidney disease in a receptor for advanced glycation end products-dependent manner[J].Arterioscler Thromb Vasc Biol,2014,34(7):1399-1411.
[7]张红欣,韩鲁军,安召伟,等.还原型谷胱甘肽对表阿霉素所致心脏毒性的保护作用及机制探讨[J].中西医结合心脑血管病杂志,2015,13(10):1219-1221.
[8]高秋,杨松,陈燕春,等.姜黄素对阿霉素致大鼠心脏毒性的保护作用及机制探讨[J].山东医药,2014,54(16):27-29.
[9]宝云龙,哈申高娃,杨莉,等.磷酸肌酸钠及丹参酮ⅡA磺酸钠对阿霉素心肌病大鼠内质网应激介导心肌细胞凋亡作用研究[J].中国分子心脏病学杂志,2014,14(5):1085-1088.
[10]王宇红,谭小雯,赵洪庆,等.左归降糖解郁方对糖尿病并发抑郁症大鼠海马星形胶质细胞GFAP、S100B的影响[J].中国中医药信息杂志,2017,24(9):1808-1811.
[11]张红心.脑苷肌肽治疗脑梗死老年患者疗效及其对血清中S100B、MMP-7和MMP-8的影响[J].重庆医学,2014(21):2781-2782.
[12]韩尧辉,潘皓,毛俊倩,等.S100B在多柔比星诱导心肌细胞损伤中的表达及作用[J].江苏大学学报(医学版),2013,23(3):216-219.
[13]Tsoporis JN,Mohammadzadeh F,Parker TG.S100B:a multifunctional role in cardiovascular pathophysiology[J].Amino Acids,2011,41(4):843-847.
[14]Villarreal A,Aviles Reyes RX,Angelo MF,et al.S100B alters neuronal survival and dendrite extension via RAGE-mediated NF-κB signaling[J].J Neurochem,2011,11 7(2):321-332.
[15]Ing DJ,Zang J,Dzau VJ,et al.Modulation of cytokineinduced cardiac myocyte apoptosis by nitric oxide,Bak,and Bcl-x[J].Circulation Res,1999,84(1):21-33.
[2]黄俊辉,张曦蓓,肖佳,等.米非司酮逆转人乳腺癌细胞MCF-7/ADR耐多柔比星机制的探讨[J].实用肿瘤杂志,2011,26(6):584-588.
[3]杜天幸,杨继元.蒽环类抗肿瘤药物对心脏毒性防治的研究进展[J].华南国防医学杂志,2015,29(1):80-82.
[4]马军,沈志祥,秦叔逵.防治蒽环类抗肿瘤药物心脏毒性的中国专家共识(2011版)[J].临床肿瘤学杂志,2011,16(12):11 22-11 29.
[5]Tsoporis J N,Marks A,Haddad A,et al.S100B expression modulates left ventricular remodeling after myocardial infarction in mice[J].Circulation,2005,111(5):598-606.
[6]Yan L,Mathew L,Chellan B,et al.S100/calgranulin-mediated inflammation accelerates left ventricular hypertrophy and aortic valve sclerosis in chronic kidney disease in a receptor for advanced glycation end products-dependent manner[J].Arterioscler Thromb Vasc Biol,2014,34(7):1399-1411.
[7]张红欣,韩鲁军,安召伟,等.还原型谷胱甘肽对表阿霉素所致心脏毒性的保护作用及机制探讨[J].中西医结合心脑血管病杂志,2015,13(10):1219-1221.
[8]高秋,杨松,陈燕春,等.姜黄素对阿霉素致大鼠心脏毒性的保护作用及机制探讨[J].山东医药,2014,54(16):27-29.
[9]宝云龙,哈申高娃,杨莉,等.磷酸肌酸钠及丹参酮ⅡA磺酸钠对阿霉素心肌病大鼠内质网应激介导心肌细胞凋亡作用研究[J].中国分子心脏病学杂志,2014,14(5):1085-1088.
[10]王宇红,谭小雯,赵洪庆,等.左归降糖解郁方对糖尿病并发抑郁症大鼠海马星形胶质细胞GFAP、S100B的影响[J].中国中医药信息杂志,2017,24(9):1808-1811.
[11]张红心.脑苷肌肽治疗脑梗死老年患者疗效及其对血清中S100B、MMP-7和MMP-8的影响[J].重庆医学,2014(21):2781-2782.
[12]韩尧辉,潘皓,毛俊倩,等.S100B在多柔比星诱导心肌细胞损伤中的表达及作用[J].江苏大学学报(医学版),2013,23(3):216-219.
[13]Tsoporis JN,Mohammadzadeh F,Parker TG.S100B:a multifunctional role in cardiovascular pathophysiology[J].Amino Acids,2011,41(4):843-847.
[14]Villarreal A,Aviles Reyes RX,Angelo MF,et al.S100B alters neuronal survival and dendrite extension via RAGE-mediated NF-κB signaling[J].J Neurochem,2011,11 7(2):321-332.
[15]Ing DJ,Zang J,Dzau VJ,et al.Modulation of cytokineinduced cardiac myocyte apoptosis by nitric oxide,Bak,and Bcl-x[J].Circulation Res,1999,84(1):21-33.