摘要
目的探讨ATP敏感钾通道(KATP)在外源性硫化氢(H2S)诱导人子宫肌瘤细胞凋亡中的作用。方法用硫氢化钠(NaHS)作为H2S的供体处理原代培养的人子宫肌瘤细胞,KATP通道抑制剂格列本脲(Gly)预处理细胞。实验分为对照组、不同浓度NaHS组、ATP敏感性钾通道(KATP)抑制剂组和Gly+NaHS组。用流式细胞术检测细胞凋亡率,采用实时定量PCR和Western blot分别检测p53和bcl-2 mRNA和蛋白的表达。结果与对照组比较,NaHS(10-4和10-3mol/L)处理人子宫肌瘤细胞24和48 h后以剂量和时间依赖的方式增加了细胞的凋亡率(均P<0.05)。与对照组比较,NaHS(10-4mol/L)处理人子宫肌瘤细胞24 h后p53的表达显著增加(P<0.05),而bcl-2表达显著降低(P<0.05)。Gly没有影响人子宫肌瘤细胞的凋亡率、p53和bcl-2的表达,但部分地拮抗NaHS的致凋亡作用以及NaHS诱导的子宫肌瘤细胞中p53表达的上调和bcl-2表达的下调(均P<0.05)。结论外源性H2S诱导子宫肌瘤细胞凋亡,其机制与H2S开放KATP通道有关。
Objective To explore the role of ATP-sensitive K＋ channels in the apoptosis induced by extrogenous hydrogen sulfide（H2S） in human uterine leiomyoma cells.Methods The human uterine leiomyoma cells were treated with a H2S donor sodium bisulfide（NaHS）.The inhibitor of ATP-sensitive K＋ channels glybenclamide（Gly） was used.The cells were divided into control,the different concentration NaHS,Gly and NaHS ＋ Gly groups.The rates of apoptosis of cells were measured by flow cytometry.The mRNA and protein expressions of p53 and bcl-2 in human leimyoma cells were measured by Real-time PCR and Western-Blot.Results Compared with the control,the rates of apoptosis were significantly increased in human uterine leiomyoma cells by treatment with NaHS（10-4 and 10-3mol/L） for 24 and 48 h in a dose-dependent and time-dependent manner.Compared with the control,the expression of p53 was significantly up-regulated and expression of bcl-2 was significantly down-regulated by treatment with NaHS（10-4mol/L） for 24 h.Gly did not affect the rates of apoptosis,expression of p53 and bcl-2 in human uterine leiomyoma cells.Gly partly blocked the effect of H2S which induced the apoptosis,up-regulated the expression of p53 and down-regulated the expression of bcl-2.Conclusions Extrogenous H2S promotes the apoptosis in human uterine leiomyoma cells,which is related with the activation of ATP-sensitive K ＋ channels by H2S.
出处
《中南医学科学杂志》
CAS
2013年第3期238-242,共5页
Medical Science Journal of Central South China
