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 cisplatin at 20 mg/L concentrations for 0,6,12,24,36,48 h respectively.HMMSCs were pretreated with NaHS at 0,0.4,0.6,0.8,1.0 mmol/L respectively for 30 min before exposed 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 NaHS 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 cisplatin 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％ after HMMSCs were treated with cisplatin at 20 mg/L for 6,12,24,36,48 h respectively,and the differences were statistically significant 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 NaHS,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 NaHS 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 NaHS could inhibit the inhibitory action to the expression of p-ERK1/2 induced by cisplatin.Pretreatment with U0126 or HEGF inhibited or promoted the protection and the upregulated expression ofp-ERK1/2 caused by NaHS pretreatment.Conclusion The preconditioning of H2S can protect cell damage caused by cisplatin via activating the ERK1/2 pathway of HMMSCs.%目的 探讨硫化氢在人骨髓间质干细胞(human marrow mesenchymal stem cells,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、80 mg/L顺铂分别作用HMMSCs 24 h;20 mg/L顺铂分别作用0、6、12、24、36、48 h；在20 mg/L顺铂作用24 h前分别用0、0.4、0.6、0.8、1.0 mmol/L的NaHS预处理30 min; 0.6 mmol/L的NaHS预处理30 min前用ERK1/2通路抑制剂U0126处理30 min或用激动剂人上皮生长因子共同预处理30 min.检测细胞存活率、细胞凋亡率、磷酸化ERK 1/2(p-ERK1/2)和总ERK1/2(t-ERK1/2)的表达.结果 5、10、20、40、60、80 mg/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0mg/L组的差异有统计学意义；20 mg/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；0.6 mmol/L的NaHS预处理使细胞凋亡率从35.29％±2.77％下降至18.62％±0.97％；20 mg/L顺铂作用24 h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.
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