ERK1／2通路介导硫化氢在入骨髓间质干细胞对抗顺铂诱导损伤中的作用  

ERK1/2 pathway mediates the protective effect of hydrogen sulfide against cisplatin-induced injury in human marrow mesenchymai stem cells

作　　者：郭海华[1] 黄纲[1] 李敬春 冯鉴强[3] 雍碧城[1] 

机构地区：[1]中山大学附属第一医院骨肿瘤科,广州510080 [2]广州市妇女儿童医疗中心骨科 [3]中山大学中医学院生理教研室

出　　处：《中华骨科杂志》2014年第3期323-329,共7页Chinese Journal of Orthopaedics

基　　金：广东省科技计划项目（20128031800050）

摘　　要：目的探讨硫化氢在人骨髓间质干细胞（humanmarrowmesenchymalstemcells，HMMSCs）对抗顺铂诱导损伤中的作用及其机制。方法0、5、10、20、40、60、80mg／L顺铂分别作用HMMSCs24h；20mg／L顺铂分别作用0、6、12、24、36、48h；在20mg／L顺铂作用24h前分别用0、0．4、0．6、0．8、1．0mmol／L的NaHS预处理30min；0．6mmol／L的NaHS预处理30min前用ERK1／2通路抑制剂U0126处理30min或用激动剂人上皮生长因子共同预处理30min。检测细胞存活率、细胞凋亡率、磷酸化ERK1／2（p—ERK1／2）和总ERK1／2（t．ERK1／2）的表达。结果5、10、20、40、60、80mg／L顺铂作用后细胞存活率分别降至71．72％±2．72％、59．41％±5．44％、50-37％±4．55％、38．97％±2．92％、30．11％±4．64％、21．71％±5．35％，与0mglL组的差异有统计学意义；20mg／L顺铂作用不同时间后细胞存活率分别降至70．30％±6．20％、61．63％±2．70％、51．29％±3．13％、38．72％±3．66％、27．57％±2．32％，与0h组的差异有统计学意义；不同浓度NaHS预处理可使细胞存活率升高至65．99％±2．67％、72．93％±5．44％、75．10％±4．71％、76．56％＋5．25％，与顺铂处理组的差异有统计学意义；0．6mmol／L的NaHS预处理使细胞凋亡率从35．29％±2．77％下降至18．62％±O．97％；20mg／L顺铂作用24h使p-ERK1/2表达下调，NaHS预处理可抑制顺铂对p-ERK1/2表达的抑制作用。应用U0126或人上皮生长因子分别抑制或促进了NaHS的保护作用及上调p-ERK1/2表达的作用。结论硫化氢预处理能通过激活HMMSCs的ERK1／2通路对抗顺铂诱导的细胞损伤。Objective To explore the protection and mechanism of hydrogen sulfide （H2S） preconditioning against injury induced by cisplatin in human marrow mesenchymal stem cells （HMMSCs）. Methods HMMSCs were treated by cisplatin at 0, 5, 10, 20, 40, 60, 80 mg/L concentrations for 24 h respectively and were exposed to eisplatin at 20 mg/L concentrations for 0, 6, 12, 24, 36, 48 h respectively. HMMSCs were pretreated with NariS at 0, 0.4, 0.6, 0.8, 1.0 mmol/L respectively for 30 min before ex- posed to cisplatin at 20 mg/L concentrations for 24 h. HMMSCs were treated by U0126 or combined with human epidermal growth factor （HEGF） together for 30 min before they were preconditioned with NariS for 30 min. The cell survival rate, cell apoptosis rate, the expression of p-ERK1/2 and t-ERK1/2 were recorded. Results The cell survival rate decreased to 71.72%±2.72%, 59.41%±5.44%, 50.37%±4.55%, 38.97%±2.92%, 30.11%±4.64% and 21.71%_±5.35% respectively after cells were treated with eisplatin at 5, 10, 20, 40, 60, 80 mg/L concentrations respectively, and the differences were statistically significant compared with 0 mg/L group. The cell viability fell to 70.30%_±6.20%, 61.63%±2.70%, 51.29%_±3.13%, 38.72%±3.66% and 27.57%_±2.32% af- ter HMMSCs were treated with cisplatin at 20 mg/L for 6, 12, 24, 36, 48 h respectively, and the differences were statistically signif- icant compared with 0 h group. The cell viability increased to 65.99%±2.67%, 72.93%_±5.44%, 75.10%±4.71% and 76.56%-± 5.25% when HMMSCs got pretreatment of NariS, and the differences had statistical significance compared with cisplatin group. The cell apoptotic rate decreased from 35.29%±2.77% to 18.62%±0.97% when HMMSCs were pretreated with Naris at 0.6 mmo/ L. Treatment of HMMSCs with cisplatin at 20 mg/L for 24 h reduced p-ERK1/2 expression. The pretreatment of Naris could inhib- it the inhibitory action to the expression of p-ERK1/2 induced by eisplatin. Pretreatment with U0126 or HEGF inhibited or promot- ed the protection and the upregulated e

关 键 词：MAP激酶信号系统 硫化氢 间质干细胞 顺铂 细胞保护 

分 类 号：R363[医药卫生—病理学]