中华骨科杂志
中華骨科雜誌
중화골과잡지
CHINESE JOURNAL OF ORTHOPAEDICS
2014年
3期
323-329
,共7页
郭海华%黄纲%李敬春%冯鉴强%雍碧城
郭海華%黃綱%李敬春%馮鑒彊%雍碧城
곽해화%황강%리경춘%풍감강%옹벽성
MAP激酶信号系统%硫化氢%间质干细胞%顺铂%细胞保护
MAP激酶信號繫統%硫化氫%間質榦細胞%順鉑%細胞保護
MAP격매신호계통%류화경%간질간세포%순박%세포보호
MAP kinase signaling system%Hydrogen sulfide%Mesenchymal stem cells%Cisplatin%Cytoprotection
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.
目的 探討硫化氫在人骨髓間質榦細胞(human marrow mesenchymal stem cells,HMMSCs)對抗順鉑誘導損傷中的作用及其機製.方法 0、5、10、20、40、60、80 mg/L順鉑分彆作用HMMSCs 24 h;20 mg/L順鉑分彆作用0、6、12、24、36、48 h；在20 mg/L順鉑作用24 h前分彆用0、0.4、0.6、0.8、1.0 mmol/L的NaHS預處理30 min; 0.6 mmol/L的NaHS預處理30 min前用ERK1/2通路抑製劑U0126處理30 min或用激動劑人上皮生長因子共同預處理30 min.檢測細胞存活率、細胞凋亡率、燐痠化ERK 1/2(p-ERK1/2)和總ERK1/2(t-ERK1/2)的錶達.結果 5、10、20、40、60、80 mg/L順鉑作用後細胞存活率分彆降至71.72％±2.72％、59.41％±5.44％、50.37％±4.55％、38.97％±2.92％、30.11％±4.64％、21.71％±5.35％,與0mg/L組的差異有統計學意義；20 mg/L順鉑作用不同時間後細胞存活率分彆降至70.30％±6.20％、61.63％±2.70％、51.29％±3.13％、38.72％±3.66％、27.57％±2.32％,與0h組的差異有統計學意義；不同濃度NaHS預處理可使細胞存活率升高至65.99％±2.67％、72.93％±5.44％、75.10％±4.71％、76.56％±5.25％,與順鉑處理組的差異有統計學意義；0.6 mmol/L的NaHS預處理使細胞凋亡率從35.29％±2.77％下降至18.62％±0.97％；20 mg/L順鉑作用24 h使p-ERK1/2錶達下調,NaHS預處理可抑製順鉑對p-ERK1/2錶達的抑製作用.應用U0126或人上皮生長因子分彆抑製或促進瞭NaHS的保護作用及上調p-ERK1/2錶達的作用.結論 硫化氫預處理能通過激活HMMSCs的ERK1/2通路對抗順鉑誘導的細胞損傷.
목적 탐토류화경재인골수간질간세포(human marrow mesenchymal stem cells,HMMSCs)대항순박유도손상중적작용급기궤제.방법 0、5、10、20、40、60、80 mg/L순박분별작용HMMSCs 24 h;20 mg/L순박분별작용0、6、12、24、36、48 h；재20 mg/L순박작용24 h전분별용0、0.4、0.6、0.8、1.0 mmol/L적NaHS예처리30 min; 0.6 mmol/L적NaHS예처리30 min전용ERK1/2통로억제제U0126처리30 min혹용격동제인상피생장인자공동예처리30 min.검측세포존활솔、세포조망솔、린산화ERK 1/2(p-ERK1/2)화총ERK1/2(t-ERK1/2)적표체.결과 5、10、20、40、60、80 mg/L순박작용후세포존활솔분별강지71.72％±2.72％、59.41％±5.44％、50.37％±4.55％、38.97％±2.92％、30.11％±4.64％、21.71％±5.35％,여0mg/L조적차이유통계학의의；20 mg/L순박작용불동시간후세포존활솔분별강지70.30％±6.20％、61.63％±2.70％、51.29％±3.13％、38.72％±3.66％、27.57％±2.32％,여0h조적차이유통계학의의；불동농도NaHS예처리가사세포존활솔승고지65.99％±2.67％、72.93％±5.44％、75.10％±4.71％、76.56％±5.25％,여순박처리조적차이유통계학의의；0.6 mmol/L적NaHS예처리사세포조망솔종35.29％±2.77％하강지18.62％±0.97％；20 mg/L순박작용24 h사p-ERK1/2표체하조,NaHS예처리가억제순박대p-ERK1/2표체적억제작용.응용U0126혹인상피생장인자분별억제혹촉진료NaHS적보호작용급상조p-ERK1/2표체적작용.결론 류화경예처리능통과격활HMMSCs적ERK1/2통로대항순박유도적세포손상.
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.
