S1P对缺氧/复氧诱导的共培养心肌细胞和成肌细胞损伤的影响  

Effects of S1P on Anoxia/Reoxygenation Injury in Co-cultured Cardiomyocytes and Myoblasts

作　　者：于欢[1] 谷翔[2] 张普华[2] 徐良贤[1] 陈培良[1] 秦仲君 赵勇[1] 

机构地区：[1]九江学院江西省系统生物医学重点实验室,九江332000 [2]九江学院附属医院心内科,九江332000

出　　处：《华中科技大学学报（医学版）》2013年第6期637-641,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家青年科学基金资助项目(No.81000075);江西省教育厅重点科研项目(No.GJJ11694);江西省教育厅科研项目(No.MP876;No.GJJ11242);江西省卫生厅科研项目(No.20113138)

摘　　要：目的探讨1-磷酸神经鞘氨醇（sphingosine 1-phosphate,S1P）对联合培养的大鼠骨骼肌成肌细胞及新生大鼠心肌细胞缺氧/复氧所致细胞损伤的影响。方法体外分离与培养新生大鼠心肌细胞,并与大鼠骨骼肌成肌细胞共同培养,构建缺氧/复氧损伤模型,于复氧期开始给予S1P干预。利用2,4二硝基苯肼显色法检测乳酸脱氢酶（lactate dehydrogenase,LDH）活性,硫代巴比妥酸显色法检测丙二醛（maleic dialdehyde,MDA）含量,原位末端标记法（TUNEL）检测细胞凋亡,并比较各组间差异,从而观察S1P对共同培养新生大鼠心肌细胞和大鼠骨骼肌成肌细胞缺氧/复氧诱导细胞损伤的影响。结果①新生大鼠心肌细胞在培养24h后开始贴壁生长,培养6d内细胞数量未见明显变化,培养2d后倒置显微镜下见散在的具有搏动能力的心肌细胞,第3天可见散在的心肌细胞相互间连接成合胞且可见细胞搏动,4~5d呈同一节律的搏动。②100、500、1 000nmol/L S1P均可减少共培养心肌细胞和骨骼肌成肌细胞缺氧/复氧损伤后LDH活性和MDA含量,即可减轻复氧所诱导的细胞损伤。③S1P可抑制共培养心肌细胞和骨骼肌成肌细胞缺氧/复氧损伤后的细胞凋亡。结论 S1P可减少因复氧所诱导的共同培养细胞损伤,抑制细胞凋亡。Objective To explore the effects of sphingosine 1-phosphate（S1P）on the anoxia/reoxygenation（A/R）injury in co-cultured rat skeletal myoblasts and neonatal rat cardiomyocytes.Methods The neonatal rat cardiomyocytes were isolated in vitro and cultured.They were then co-cultured with rat skeletal myoblasts.The A/R injury model was established and S1P was given at the beginning of the reoxygenation.The activity of lactate dehydrogenase（LDH）and the content of maleic dialdehyde（MDA）were measured by using spectrophotometric procedures.Apoptosis of cells was assessed by TUNEL assay.The effects of S1Pon the A/R injury in co-cultured cells were examined.Results ① The neonatal rat cardiocmyocytes began to adhere after 24hof culture.The cell number experienced no significant change within 6days of culture.Beating cardiomyocytes were sparsely seen after 2days of culture under the inverse microscope.The disperse cardiomyocytes were connected on the 3rd day and beat on the same rhythm at 4-5days.② S1P（100,500and 1 000nmol/L）could significantly reduce the activity of LDH and the content of MDA induced by the A/R injury in co-cultured cardiomyocytes and skeletal myoblasts,which suggested that S1Pcould alleviate the A/R-induced cell injury.③S1Pcould inhibit A/R-induced apoptosis of co-cultured cardiomyocytes and skeletal myoblasts.Conclusion S1Pcould protect cardiomyocytes from A/R-induced injury by inhibiting cell apoptosis

关 键 词：1-磷酸神经鞘氨醇 骨骼肌成肌细胞 心肌细胞 缺氧 复氧 细胞凋亡 

分 类 号：R363[医药卫生—病理学]